WO2024137663A2 - Traitement de maladies et de troubles liés au dkk2 - Google Patents

Traitement de maladies et de troubles liés au dkk2 Download PDF

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WO2024137663A2
WO2024137663A2 PCT/US2023/084879 US2023084879W WO2024137663A2 WO 2024137663 A2 WO2024137663 A2 WO 2024137663A2 US 2023084879 W US2023084879 W US 2023084879W WO 2024137663 A2 WO2024137663 A2 WO 2024137663A2
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modified
oligonucleotide
sense strand
nucleoside
nucleosides
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WO2024137663A3 (fr
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Omri GOTTESMAN
Shannon BRUSE
Paul BUSKE
Brian CAJES
David JAKUBOSKY
Sarah KLEINSTEIN
David Lewis
David Rozema
John VEKICH
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Empirico Inc
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Empirico Inc
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Publication of WO2024137663A3 publication Critical patent/WO2024137663A3/fr
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/542—Carboxylic acids, e.g. a fatty acid or an amino acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering nucleic acids [NA]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/315—Phosphorothioates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/34—Spatial arrangement of the modifications
    • C12N2310/343—Spatial arrangement of the modifications having patterns, e.g. ==--==--==--
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/35—Nature of the modification
    • C12N2310/351—Conjugate
    • C12N2310/3515—Lipophilic moiety, e.g. cholesterol
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2320/00—Applications; Uses
    • C12N2320/10—Applications; Uses in screening processes
    • C12N2320/11—Applications; Uses in screening processes for the determination of target sites, i.e. of active nucleic acids

Definitions

  • Alopecia hair loss
  • improved therapeutics are needed for treating hair loss.
  • composition comprising an oligonucleotide that targets dickkopf WNT signaling pathway inhibitor 2 (DKK2) and when administered to a subject in an effective amount increases a hair count in the subject wherein the oligonucleotide is selected from SEQ ID NO: 7770-7821.
  • the hair count is increased by about 10% or more, as compared to prior to administration.
  • the hair count includes a vellus hair count, a non-vellus hair count, or a total hair count.
  • composition comprising an oligonucleotide that targets dickkopf WNT signaling pathway inhibitor 2 (DKK2) and when administered to a subject in an effective amount increases a hair thickness measurement in the subject wherein the oligonucleotide is selected from SEQ ID NO: 7770-7821.
  • the hair thickness measurement is increased by about 10% or more, as compared to prior to administration.
  • composition comprising an oligonucleotide that targets dickkopf WNT signaling pathway inhibitor 2 (DKK2) and when administered to a subject in an effective amount increases a hair density measurement in the subject wherein the oligonucleotide is selected from SEQ ID NO: 7770-7821.
  • the hair density measurement is increased by about 10% or more, as compared to prior to administration.
  • composition comprising an oligonucleotide that targets dickkopf WNT signaling pathway inhibitor 2 (DKK2) and when administered to a subject in an effective amount increases a number of hair follicles in the subject wherein the oligonucleotide is selected from SEQ ID NO: 7770-7821.
  • the number of hair follicles is increased by about 10% or more, as compared to prior to administration.
  • the number of hair follicles includes a number of terminal hair follicles, a number of anagen hair follicles, a number of telogen hair follicles, a number of catagen hair follicles, a number of vellus-like miniaturized hair follicles, a number of indeterminate hair follicles, or a total number of hair follicles.
  • composition comprising an oligonucleotide that targets dickkopf WNT signaling pathway inhibitor 2 (DKK2) and when administered to a subject in an effective amount changes a hair loss assessment score in the subject wherein the oligonucleotide is selected from SEQ ID NO: 7770-7821.
  • the hair loss assessment score is changed by about 10% or more, as compared to prior to administration.
  • composition comprising an oligonucleotide that targets dickkopf WNT signaling pathway inhibitor 2 (DKK2) and when administered to a subject in an effective amount changes a protein or mRNA level of [3-catenin, a-SMA, collagen I, or collagen III, in the subject wherein the oligonucleotide is selected from SEQ ID NO: 7770-7821.
  • the protein or mRNA level of [3-catenin, a-SMA, collagen I, or collagen III is changed by about 10% or more, as compared to prior to administration.
  • composition comprising an oligonucleotide that targets dickkopf WNT signaling pathway inhibitor 2 (DKK2) and when administered to the skin of a subject in an effective amount decreases a level of DKK2 mRNA or DKK2 protein wherein the oligonucleotide is selected from SEQ ID NO: 7770-7821.
  • the skin comprises scalp skin.
  • the level of DKK2 mRNA or DKK2 protein decreased by about 10% or more, as compared to prior to administration.
  • the oligonucleotide comprises a modified intemucleoside linkage.
  • the modified intemucleoside linkage comprises alkylphosphonate, phosphorothioate, methylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, or carboxymethyl ester, or a combination thereof.
  • the modified intemucleoside linkage comprises one or more phosphorothioate or phosphate linkages.
  • the oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 modified intemucleoside linkages.
  • the oligonucleotide comprises 2 or more modified intemucleoside linkages, 3 or more modified intemucleoside linkages, 4 or more modified intemucleoside linkages, 5 or more modified intemucleoside linkages, 6 or more modified intemucleoside linkages, 7 or more modified intemucleoside linkages, 8 or more modified intemucleoside linkages, 9 or more modified intemucleoside linkages, 10 or more modified intemucleoside linkages, 11 or more modified intemucleoside linkages, 12 or more modified intemucleoside linkages, 13 or more modified intemucleoside linkages, 14 or more modified intemucleoside linkages, 15 or more modified intemucleoside linkages, 16 or more modified intemucleoside linkages, 17 or more modified intemucleoside linkages, 18 or more modified intemucleoside linkages, 19 or more modified intemucleoside linkages, or 20 or more modified in
  • the oligonucleotide comprises a modified nucleoside.
  • the modified nucleoside comprises a locked nucleic acid (LNA), hexitol nucleic acid (HNA), cyclohexene nucleic acid (CeNA), 2'-O-methoxyethyl, 2'-O-alkyl, 2'-O-allyl, 2'-O-allyl, 2'-fluoro, or 2'-deoxy, or a combination thereof.
  • the modified nucleoside comprises an LNA.
  • the modified nucleoside comprises a 2’, 4’ constrained ethyl nucleic acid.
  • the modified nucleoside comprises a 2'-O-methyl nucleoside, 2'-deoxyfluoro nucleoside, 2'-O-N-methylacetamido (2'-0-NMA) nucleoside, a 2'-O- dimethylaminoethoxyethyl (2'-0-DMAE0E) nucleoside, 2'-O-aminopropyl (2'-O-AP) nucleoside, or 2'-ara-F, or a combination thereof.
  • the modified nucleoside comprises one or more 2’-fluoro modified nucleosides.
  • the modified nucleoside comprises a 2'-O-alkyl modified nucleoside.
  • the oligonucleotide comprises a lipid attached at a 3’ or 5’ terminus of the oligonucleotide.
  • the lipid comprises cholesterol, myristoyl, palmitoyl, stearoyl, lithocholoyl, docosanoyl, docosahexaenoyl, myristyl, palmityl, stearyl, or a-tocopherol, or a combination thereof.
  • the oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 modified nucleosides.
  • the oligonucleotide comprises 2 or more modified nucleosides, 3 or more modified nucleosides, 4 or more modified nucleosides, 5 or more modified nucleosides, 6 or more modified nucleosides, 7 or more modified nucleosides, 8 or more modified nucleosides, 9 or more modified nucleosides, 10 or more modified nucleosides, 11 or more modified nucleosides, 12 or more modified nucleosides, 13 or more modified nucleosides, 14 or more modified nucleosides, 15 or more modified nucleosides, 16 or more modified nucleosides, 17 or more modified nucleosides, 18 or more modified nucleosides, 19 or more modified nucleosides, 20 or more modified nucleosides, or 21 or more modified nucleosides.
  • the oligonucleotide comprises a small interfering RNA (siRNA) comprising a sense strand and an antisense strand.
  • siRNA small interfering RNA
  • the sense strand is 14-30 nucleosides in length.
  • the antisense strand is 14-30 nucleosides in length.
  • composition comprising an oligonucleotide that inhibits the expression of dickkopf WNT signaling pathway inhibitor 2 (DKK2) wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, each strand is independently about 14-30 nucleosides in length, and at least one of the sense strand and the antisense strand comprises a nucleoside sequence comprising about 14-30 contiguous nucleosides of SEQ ID NO: 1 wherein the sense strand is selected from SEQ ID NO: 7770-7821, and wherein the antisense strand is selected from SEQ ID NO: 7822-7873.
  • DKK2 dickkopf WNT signaling pathway inhibitor 2
  • composition comprising an oligonucleotide that inhibits the expression of dickkopf WNT signaling pathway inhibitor 2 (DKK2), wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, each strand is independently about 14-30 nucleosides in length, and at least one of the sense strand and the antisense strand comprises a nucleoside sequence comprising about 14-30 contiguous nucleosides of a full-length human DKK2 pre- mRNA sequence wherein the sense strand is selected from SEQ ID NO: 7770-7821 and wherein the antisense strand is selected from SEQ ID NO: 7822-7873.
  • DKK2 dickkopf WNT signaling pathway inhibitor 2
  • the sense strand is selected from the group consisting of SEQ ID NO: 7787, SEQ ID NO: 7799, and SEQ ID NO: 7804; wherein the antisense strand is selected from the group consisting of SEQ ID NO: 7839, SEQ ID NO: 7821, and SEQ ID NO: 7856.
  • the sense strand is selected from the group consisting of: SEQ ID NO: 7684, SEQ ID NO: 7874, and SEQ ID NO: 7875; wherein the antisense strand is selected from the group consisting of: SEQ ID NO: 7328, SEQ ID NO: 7747, and SEQ ID NO: 7752.
  • the oligonucleotide further comprises a modification pattern.
  • the modification pattern is selected from the group consisting of: 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, 68S, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, I 3AS.
  • N is any nucleotide
  • dN is a 2’ deoxy-modified nucleoside
  • Nf is a 2’-fluoro-modified nucleoside
  • n is a 2’-O-methyl modified nucleoside
  • nm is a 2’-O-methoxyethyl modified nucleoside
  • s is a phosphorothioate or phosphate linkage.
  • composition comprising a small interfering RNA (siRNA) that targets dickkopf WNT signaling pathway inhibitor 2 (DKK2) and when administered to a cell decreases expression of DKK2, wherein the siRNA comprises a sense strand and an antisense strand; and wherein the antisense strand comprises modification pattern 6AS, 7AS, 8AS, 9AS, 10AS, 1 IAS, 12AS, HAS.
  • siRNA small interfering RNA
  • DKK2 dickkopf WNT signaling pathway inhibitor 2
  • the sense strand comprises a modification pattern selected from the group consisting of: 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 5 IS, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, and 68S, wherein “Nf ’ is a 2’-fluoro- modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “nm” is a 2’-O-methoxyethyl modified nucleoside and “s” is a phosphorothioate or phosphate linkage.
  • a method of treating hair loss in a subject in need thereof comprising administering to the subject a composition described herein.
  • the hair loss comprises any one or more of male pattern baldness, female pattern baldness, alopecia areata, or non-scarring hair loss.
  • the administration is topical.
  • FIG. 1 depicts a plot of DKK2 mRNA expression in Lymphoblastoid Cell Lines from donors with known rs35290077 (G96R) genotypes following treatment with Vitamin D (dark, left) or vehicle (light, right).
  • the plot shows each copy of the rs35290077 alternative (G) allele results in approximately 50% reduction of DKK2 mRNA induction in the presence of Vitamin D.
  • the x-axis is labeled rs35290077 Donor genotype for (left to right) C/C/, C/G, and G/G/; the y-axis is labeled relative DKK2 expression from 0 to 4 at 1 unit intervals.
  • FIG. 2A depicts images of 2 mice 14 days following hair removal, and after topical treatment with a negative control siRNA (ETD01043). The images show whitening and graying of the fur on the backs of the mice in the hair that grew back.
  • ETD01043 negative control siRNA
  • FIG. 2B depicts images of 2 mice 14 days following hair removal, and after topical treatment with a DKK2 siRNA (ETD01551). The images indicate that coat color of the mice was retained in the hair that grew back on these mice.
  • DKK2 siRNA ETD01551
  • a Genome Wide Association Study detects associations between genetic variants and traits in a population sample and this improves understanding of the biology of disease and provides evidence of applicable treatments.
  • a GWAS generally utilizes genotyping and/or sequencing data, and often involves an evaluation of millions of genetic variants that are relatively evenly distributed across the genome.
  • the most common GWAS design is the case-control study, which involves comparing variant frequencies in cases versus controls. If a variant has a significantly different frequency in cases versus controls, that variant is considered associated with disease.
  • Association statistics used in a GWAS include p-values, as a measure of statistical significance; odds ratios (OR), as a measure of effect size; or beta coefficients (beta), as a measure of effect size.
  • OR odds ratios
  • beta beta coefficients
  • An additional concept in design and interpretation of GWAS is that of linkage disequilibrium, which is the non-random association of alleles. The presence of linkage disequilibrium can obfuscate which variant is “causal.”
  • Hair loss is a common problem, particularly among men, but is also common among women. It may be affected by any of several factors, including heredity, hormones, thyroid disorders, nutritional status, environmental factors, physical stress, or emotional stress. Hair loss may include, among other things, androgenetic alopecia (male pattern baldness), alopecia areata, or non-scarring hair loss.
  • the DKK2 gene is located on chromosome 4 and encodes dickkopf WNT signaling pathway inhibitor 2 (DKK2), a member of the dickkopf family.
  • DKK2 dickkopf WNT signaling pathway inhibitor 2
  • a non-limiting example of a DKK2 gene is included on GenBank under NCBI reference sequence NM_014421.3 (May 9, 2020).
  • DKK2 proteins may be secreted, include two cysteine rich regions, and be involved in embryonic development through interactions with the Wnt signaling pathway. DKK2 can act as either an agonist or antagonist of Wnt/beta-catenin signaling, depending on the cellular context and the presence of the co-factor kremen 2. Activity of DKK2 may also be modulated by binding to the Wnt co-receptor LDL-receptor related protein 6 (LRP6). In some cases, DKK2 protein is intracellular. In some cases, DKK2 protein is secreted. The secreted DKK2 protein may be locally secreted. Here, it is shown that genetic variants that cause inactivation of the DKK2 gene in humans are associated with decreased risk of male pattern baldness. Therefore, inhibition of DKK2 serve as a therapeutic strategy for treatment of hair loss such as male pattern baldness, alopecia areata, or non-scarring hair loss.
  • LRP6 Wnt co-receptor LDL-receptor
  • compositions comprising an oligonucleotide that targets DKK2.
  • the oligonucleotide may include a small interfering RNA (siRNA) or an antisense oligonucleotide (ASO).
  • siRNA small interfering RNA
  • ASO antisense oligonucleotide
  • methods of treating hair loss by providing an oligonucleotide that targets DKK2 to a subject in need thereof.
  • compositions comprising an oligonucleotide.
  • the composition comprises an oligonucleotide that targets dickkopf WNT signaling pathway inhibitor 2 (DKK2).
  • DKK2 dickkopf WNT signaling pathway inhibitor 2
  • the composition consists of an oligonucleotide that targets DKK2.
  • the oligonucleotide reduces DKK2 mRNA expression in the subject.
  • the oligonucleotide reduces DKK2 protein expression in the subject.
  • the oligonucleotide may include a small interfering RNA (siRNA) described herein.
  • the oligonucleotide may include an antisense oligonucleotide (ASO) described herein.
  • ASO antisense oligonucleotide
  • a composition described herein is used in a method of treating a disorder in a subject in need thereof.
  • Some embodiments relate to a composition comprising an oligonucleotide for use in a method of treating a disorder as described herein.
  • Some embodiments relate to use of a composition comprising an oligonucleotide, in a method of treating a disorder as described herein.
  • Some embodiments include a composition comprising an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount decreases DKK2 mRNA or protein levels in a cell, fluid or tissue. Some embodiments include a composition comprising an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount decreases DKK2 mRNA levels in a cell or tissue.
  • the cell is a skin cell.
  • the tissue is skin (e.g. scalp dermis).
  • the DKK2 mRNA levels are decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration.
  • the DKK2 mRNA levels are decreased by about 10% or more, as compared to prior to administration. In some embodiments, the DKK2 mRNA levels are decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the DKK2 mRNA levels are decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the DKK2 mRNA levels are decreased by no more than about 10%, as compared to prior to administration.
  • the DKK2 mRNA levels are decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the DKK2 mRNA levels are decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or by a range defined by any of the two aforementioned percentages.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount decreases DKK2 protein levels in a cell, fluid or tissue.
  • the cell is a skin cell.
  • the fluid is a blood, serum, or plasma.
  • the administration of the oligonucleotide decreases circulatingDKK2 protein levels.
  • the tissue is skin (e.g. scalp dermis).
  • the DKK2 protein levels are decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration.
  • the DKK2 protein levels are decreased by about 10% or more, as compared to prior to administration. In some embodiments, the DKK2 protein levels are decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the DKK2 protein levels are decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the DKK2 protein levels are decreased by no more than about 10%, as compared to prior to administration.
  • the DKK2 protein levels are decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the DKK2 protein levels are decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or by a range defined by any of the two aforementioned percentages.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount decreases hair loss (e.g. male pattern baldness, alopecia areata, or non-scarring hair loss) or a symptom of hair loss.
  • hair loss e.g. male pattern baldness, alopecia areata, or non-scarring hair loss
  • the hair loss or symptom of hair loss is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration.
  • the hair loss or symptom of hair loss is decreased by about 10% or more, as compared to prior to administration.
  • the hair loss or symptom of hair loss is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100%, as compared to prior to administration. In some embodiments, the hair loss or symptom of hair loss is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the hair loss or symptom of hair loss is decreased by no more than about 10%, as compared to prior to administration.
  • the hair loss or symptom of hair loss is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration.
  • the hair loss or symptom of hair loss is decreased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or by a range defined by any of the two aforementioned percentages.
  • the hair loss or symptom of hair loss is assessed by phototrichogram.
  • the hair loss or symptom of hair loss is assessed by a macrophotography analysis.
  • the hair loss or symptom of hair loss is assessed by a questionnaire such as a Men's Hair Growth Questionnaire (MHGQ) or a Kingsley Alopecia Profile (KAP) questionnaire.
  • MHGQ Men's Hair Growth Questionnaire
  • KAP Kingsley Alopecia Profile
  • the hair loss or symptom of hair loss is assessed by a scalp biopsy.
  • the decrease in hair loss or symptom of hair loss is determined as a change in a hair loss hair loss assessment score.
  • the change in the hair loss assessment score may be an increase in the hair loss assessment score.
  • the change in the hair loss assessment score is a decrease in the hair loss assessment score.
  • the hair loss assessment score is obtained as part of an assessment that includes the questionnaire.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount results in a change in an hair loss assessment score.
  • the hair loss assessment score is changed by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration.
  • the hair loss assessment score is changed by about 10% or more, as compared to prior to administration.
  • the hair loss assessment score is changed by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration.
  • the hair loss assessment score is changed by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the hair loss assessment score is changed by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the hair loss assessment score is changed by no more than about 10%, as compared to prior to administration.
  • the hair loss assessment score is changed by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the hair loss assessment score is changed by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration.
  • the hair loss assessment score is changed by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount increases a total hair count (e.g. a vellus and non-vellus hair count).
  • a total hair count e.g. a vellus and non-vellus hair count.
  • the total hair count is determined in an area of skin.
  • the total hair count is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration.
  • the total hair count is increased by about 10% or more, as compared to prior to administration.
  • the total hair count is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the total hair count is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the total hair count is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration.
  • the total hair count is increased by no more than about 10%, as compared to prior to administration. In some embodiments, the total hair count is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the total hair count is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration.
  • the total hair count is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the total hair count is assessed by phototrichogram.
  • the total hair count is assessed by a macrophotography analysis.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount increases a vellus hair count.
  • the vellus hair count is determined in an area of skin.
  • the vellus hair count is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration.
  • the vellus hair count is increased by about 10% or more, as compared to prior to administration.
  • the vellus hair count is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the vellus hair count is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the vellus hair count is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration.
  • the vellus hair count is increased by no more than about 10%, as compared to prior to administration. In some embodiments, the vellus hair count is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the vellus hair count is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration.
  • the vellus hair count is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the vellus hair count is assessed by phototrichogram.
  • the vellus hair count is assessed by a macrophotography analysis.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount increases a non-vellus hair count.
  • the non-vellus hair count is determined in an area of skin.
  • the non-vellus hair count is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration.
  • the non-vellus hair count is increased by about 10% or more, as compared to prior to administration.
  • the non-vellus hair count is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the non-vellus hair count is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration.
  • the non-vellus hair count is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the non-vellus hair count is increased by no more than about 10%, as compared to prior to administration. In some embodiments, the non-vellus hair count is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration.
  • the non-vellus hair count is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration.
  • the non-vellus hair count is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the non-vellus hair count is assessed by phototrichogram.
  • the non-vellus hair count is assessed by a macrophotography analysis.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount increases hair thickness.
  • the thicknesses (e.g. widths) of individual hairs are increased.
  • the hair thickness is determined in an area of skin.
  • the increased hair thickness may include an increased vellus hair thickness.
  • the increased hair thickness may include an increased non-vellus hair thickness.
  • the hair thickness is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration.
  • the hair thickness is increased by about 10% or more, as compared to prior to administration.
  • the hair thickness is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the hair thickness is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the hair thickness is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration.
  • the hair thickness is increased by no more than about 10%, as compared to prior to administration. In some embodiments, the hair thickness is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the hair thickness is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration.
  • the hair thickness is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the hair thickness is assessed by phototrichogram.
  • the hair thickness is assessed by a macrophotography analysis.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount increases hair density.
  • the hair density is determined in an area of skin. In some embodiments, the hair density comprises a number of hairs per an area of skin.
  • the increased hair density may include an increased vellus hair density.
  • the increased hair density may include an increased non-vellus hair density.
  • the hair density is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the hair density is increased by about 10% or more, as compared to prior to administration.
  • the hair density is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the hair density is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the hair density is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration.
  • the hair density is increased by no more than about 10%, as compared to prior to administration. In some embodiments, the hair density is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the hair density is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration.
  • the hair density is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the hair density is assessed by phototrichogram.
  • the hair density is assessed by a macrophotography analysis.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount increases a number of hair follicles.
  • the number of hair follicles is determined in an area of skin.
  • the hair follicles may include terminal hair follicles, anagen hair follicles, telogen hair follicles, catagen hair follicles, vellus or vellus- like miniaturized hair follicles, or indeterminate hair follicles.
  • the hair follicles include terminal hair follicles.
  • the hair follicles include anagen hair follicles.
  • the hair follicles include telogen hair follicles. In some embodiments, the hair follicles include catagen hair follicles. In some embodiments, the hair follicles include vellus or vellus- like miniaturized hair follicles. In some embodiments, the hair follicles include indeterminate hair follicles. In some embodiments, the number of hair follicles is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the number of hair follicles is increased by about 10% or more, as compared to prior to administration.
  • the number of hair follicles is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration. In some embodiments, the number of hair follicles is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration.
  • the number of hair follicles is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the number of hair follicles is increased by no more than about 10%, as compared to prior to administration. In some embodiments, the number of hair follicles is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration.
  • the number of hair follicles is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration.
  • the number of hair follicles is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the number of hair follicles is assessed in a scalp biopsy.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount increases a hair color measurement.
  • the hair color measurement is determined on an area of skin.
  • the hair color measurement is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration.
  • the hair color measurement is increased by about 10% or more, as compared to prior to administration.
  • the hair color measurement is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration.
  • the hair color measurement is increased by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the hair color measurement is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the hair color measurement is increased by no more than about 10%, as compared to prior to administration.
  • the hair color measurement is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the hair color measurement is increased by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration.
  • the hair color measurement is increased by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the composition comprises an oligonucleotide that targets DKK2 and when administered to a subject in an effective amount results in a change in expression of a gene or protein.
  • the gene or protein may include a [3-catenin gene.
  • the gene or protein may include a [3-catenin protein.
  • the gene or protein may include an a-SMA gene.
  • the gene or protein may include an a-SMA protein.
  • the gene or protein may include a collagen I gene.
  • the gene or protein may include a collagen I protein.
  • the gene or protein may include a collagen III gene.
  • the gene or protein may include a collagen III protein.
  • the change in expression is determined in a tissue (e.g. skin), cell, or fluid sample.
  • the expression is changed by about 2.5% or more, about 5% or more, or about 7.5% or more, as compared to prior to administration. In some embodiments, the expression is changed by about 10% or more, as compared to prior to administration. In some embodiments, the expression is changed by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 100% or more, as compared to prior to administration.
  • the expression is changed by about 200% or more, about 300% or more, about 400% or more, about 500% or more, about 600% or more, about 700% or more, about 800% or more, about 900% or more, or about 1000% or more, as compared to prior to administration. In some embodiments, the expression is changed by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, as compared to prior to administration. In some embodiments, the expression is changed by no more than about 10%, as compared to prior to administration.
  • the expression is changed by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100%, as compared to prior to administration. In some embodiments, the expression is changed by no more than about 200%, no more than about 300%, no more than about 400%, no more than about 500%, no more than about 600%, no more than about 700%, no more than about 800%, no more than about 900%, or no more than about 1000%, as compared to prior to administration.
  • the expression is changed by 2.5%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 150%, 200%, 250%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • siRNAs siRNAs
  • the composition comprises an oligonucleotide that targets DKK2, wherein the oligonucleotide comprises a small interfering RNA (siRNA). In some embodiments, the composition comprises an oligonucleotide that targets DKK2, wherein the oligonucleotide comprises a small interfering RNA (siRNA) comprising a sense strand and an antisense strand.
  • siRNA small interfering RNA
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the sense strand is 12-30 nucleosides in length. In some embodiments, the sense strand is 14-30 nucleosides in length. In some embodiments, the composition comprises a sense strand that is at least about 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleosides in length, or a range defined by any of the two aforementioned numbers. In some embodiments, the composition comprises an antisense strand is 12-30 nucleosides in length.
  • the antisense strand is 14-30 nucleosides in length. In some embodiments, the composition comprises an antisense strand that is at least about 10, 11, 12, 13, 14, 15, 1516 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleosides in length, or a range defined by any of the two aforementioned numbers.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, each strand is independently about 14-30 nucleosides in length, and at least one of the sense strand and the antisense strand comprises a nucleoside sequence comprising about 14-30 contiguous nucleosides of a full-length human DKK2 pre-mRNA sequence.
  • At least one of the sense strand and the antisense strand comprise a nucleoside sequence comprising at least about 10, 11, 12, 13, 14, 15, 15, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more contiguous nucleosides of the pre-mRNA sequence.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, each strand is independently about 14-30 nucleosides in length, and at least one of the sense strand and the antisense strand comprises a nucleoside sequence comprising about 14-30 contiguous nucleosides of a full-length human DKK2 mRNA sequence such as SEQ ID NO: 7599.
  • At least one of the sense strand and the antisense strand comprise a nucleoside sequence comprising at least about 10, 11, 12, 13, 14, 15, 15, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more contiguous nucleosides of one of SEQ ID NO: 7599.
  • the siRNA may include one or more intemucleoside linkages and/or one or more nucleoside modifications. Any one of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of 7599. Any of the aforementioned siRNAs may include an antisense strand that lacks a 5’ U of an antisense sequence of one of SEQ ID NO: 7599.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the sense strand and the antisense strand form a double -stranded RNA duplex.
  • the first base pair of the double -stranded RNA duplex is an AU base pair.
  • the sense strand further comprises a 3’ overhang.
  • the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers.
  • the 3’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 3’ overhang comprises 2 nucleosides. In some embodiments, the sense strand further comprises a 5’ overhang. In some embodiments, the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers. In some embodiments, the 5’ overhang comprises 1, 2, or more nucleosides. In some embodiments, the 5’ overhang comprises 2 nucleosides.
  • the antisense strand further comprises a 3’ overhang.
  • the 3’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers.
  • the 3’ overhang comprises 1, 2, or more nucleosides.
  • the 3’ overhang comprises 2 nucleosides.
  • the antisense strand further comprises a 5’ overhang.
  • the 5’ overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleosides, or a range of nucleotides defined by any two of the aforementioned numbers.
  • the 5’ overhang comprises 1, 2, or more nucleosides.
  • the 5’ overhang comprises 2 nucleosides.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the siRNA binds with a 19mer in a human DKK2 mRNA.
  • the siRNA binds with a 12mer, a 13mer, a 14mer, a 15mer, a 16mer, a 17mer, a 18mer, a 19mer, a 20mer, a 21mer, a 22mer, a 23mer, a 24mer, or a 25mer in a human DKK2 mRNA.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the siRNA binds with a 17mer in a non -human primate DKK2 mRNA.
  • the siRNA binds with a 12mer, a 13mer, a 14mer, a 15mer, a 16mer, a 17mer, a 18mer, a 19mer, a 20mer, a 21mer, a 22mer, a 23mer, a 24mer, or a 25mer in a non-human primate DKK2 mRNA.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the siRNA binds with a 19mer in a human DKK2 mRNA, or a combination thereof.
  • the siRNA binds with a 12mer, a 13mer, a 14mer, a 15mer, a 16mer, a 17mer, and 18mer, a 19mer, a 20mer, a 21mer, a 22mer, a 23mer, a 24mer, or a 25mer in a human DKK2 mRNA.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the siRNA binds with a human DKK2 mRNA and less than or equal to 20 human off- targets, with no more than 2 mismatches in the antisense strand.
  • the siRNA binds with a human DKK2 mRNA and less than or equal to 10 human off-targets, with no more than 2 mismatches in the antisense strand.
  • the siRNA binds with a human DKK2 mRNA and less than or equal to 30 human off-targets, with no more than 2 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human DKK2 mRNA and less than or equal to 40 human off- targets, with no more than 2 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human DKK2 mRNA and less than or equal to 50 human off-targets, with no more than 2 mismatches in the antisense strand.
  • the siRNA binds with a human DKK2 mRNA and less than or equal to 10 human off-targets, with no more than 3 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human DKK2 mRNA and less than or equal to 20 human off- targets, with no more than 3 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human DKK2 mRNA and less than or equal to 30 human off-targets, with no more than 3 mismatches in the antisense strand.
  • the siRNA binds with a human DKK2 mRNA and less than or equal to 40 human off-targets, with no more than 3 mismatches in the antisense strand. In some embodiments, the siRNA binds with a human DKK2 mRNA and less than or equal to 50 human off- targets, with no more than 3 mismatches in the antisense strand.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, siRNA binds with a human DKK2 mRNA target site that does not harbor an SNP, with a minor allele frequency (MAF) greater or equal to 1% (pos. 2-18).
  • siRNA binds with a human DKK2 mRNA target site that does not harbor an SNP, with a minor allele frequency (MAF) greater or equal to 1% (pos. 2-18).
  • the MAF is greater or equal to about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20%.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA in Table 2B.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence in Table 2B.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 2B, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 2B, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence in Table 2B.
  • the sense strand or antisense strand may comprise an overhang.
  • the sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • any of the aforementioned siRNAs may include a sense strand that lacks a 3’A of a sense strand of one of the sequences of Table 2B. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of the sequences of Table 2B.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with a base sequence of an siRNA in any of Table 4-6.
  • the sense strand or antisense strand sequence comprises or consists of sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical to a base sequence of an siRNA in any of Table 4-6.
  • the sense strand or antisense strand sequence comprises or consists of the sequence of a base sequence of an siRNA in any of Table 4-6, or a sense strand or antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand sequence comprises or consists of the sequence of a base sequence of an siRNA in any of Table 4-6, or a sense strand or antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand sequence comprises or consists of a sequence 100% identical to a base sequence of an siRNA in any of Table 4-6.
  • the sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5 ’ to 3 ’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand may comprise a modification pattern described herein.
  • the sense strand or antisense strand may comprise, or may lack an overhang.
  • the sense strand or antisense strand may comprise a lipid moiety.
  • the sense strand or antisense strand may comprise a GalNAc moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3 ’A of a sense strand of one of the sequences of Tables4-6. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5 ’ U of an antisense strand sequence of any one of the sequences of T ables 4- 6.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA in Table 13B.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence in Table 13B.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 13B, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 13B, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence in Table 13B.
  • the sense strand or antisense strand may comprise an overhang.
  • the sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • any of the aforementioned siRNAs may include a sense strand that lacks a 3’A of a sense strand of one of the sequences of Table 13B. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of the sequences of Table 13B.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA in Table 13C.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence in Table 13C.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 13C, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 13C, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence in Table 13C.
  • the sense strand or antisense strand may comprise an overhang.
  • the sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • any of the aforementioned siRNAs may include a sense strand that lacks a 3’A of a sense strand of one of the sequences of Table 13C. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5 ’ U of an antisense strand sequence of any one of the sequences of Table 13C.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA in Table 18.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence in Table 18.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 18, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 18, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence in Table 18.
  • the sense strand or antisense strand may comprise an overhang.
  • the sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • any of the aforementioned siRNAs may include a sense strand that lacks a 3’A of a sense strand of one of the sequences of Table 18. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of the sequences of Table 18.
  • the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 1-3636.
  • the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 1-3636, at least 80% identical to any one of SEQ ID NOs: 1-3636, at least 85% identical to of any one of SEQ ID NOs: 1-3636, at least 90% identical to any one of SEQ ID NOs: 1-3636, or at least 95% identical to any one of SEQ ID NOs: 1-3636.
  • the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 1-3636, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 1-3636, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 1-3636.
  • the sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5 ’ to 3 ’ direction) of any of the aforementioned sequences.
  • the sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand may comprise a modification pattern described herein.
  • the sense strand may comprise an overhang.
  • the sense strand may comprise a lipid moiety.
  • the sense strand may comprise a GalNAc moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3’A of a sense strand of one of SEQ ID NO: 1-3636.
  • the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID NOs: 7770-7821.
  • the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 7770-7821, at least 80% identical to any one of SEQ ID NOs: 7770-7821, at least 85% identical to of any one of SEQ ID NOs: 7770-7821, at least 90% identical to any one of SEQ ID NOs: 7770-7821, or at least 95% identical to any one of SEQ ID NOs: 7770-7821.
  • the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 7770-7821, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 1-3636, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 7770-7821.
  • the sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand may comprise a modification pattern described herein.
  • the sense strand may comprise an overhang.
  • the sense strand may comprise a lipid moiety.
  • the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 3637-7272.
  • the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 3637-7272, at least 80% identical to any one of SEQ ID NOs: 3637-7272, at least 85% identical to of any one of SEQ ID NOs: 3637-7272, at least 90% identical to any one of SEQ ID NOs: 3637-7272, or at least 95% identical to any one of SEQ ID NOs: 3637-7272.
  • the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 3637-7272, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 3637-7272, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 3637- 7272.
  • the antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the antisense strand may comprise an overhang.
  • the antisense strand may comprise a modification pattern described herein.
  • the antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of SEQ ID NO 3637-7272.
  • the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID NOs: 7822-7873.
  • the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID NOs: 7822-7873, at least 80% identical to any one of SEQ ID NOs: 7822-7873, at least 85% identical to of any one of SEQ ID NOs: 7822-7873, at least 90% identical to any one of SEQ ID NOs: 7822-7873, or at least 95% identical to any one of SEQ ID NOs: 7822-7873.
  • the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 7822-7873, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID NOs: 7822-7873, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NOs: 7822- 7873.
  • the antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the antisense strand may comprise an overhang.
  • the antisense strand may comprise a modification pattern described herein.
  • the antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of SEQ ID NO 7822-7873.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset A.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset A.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset A, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset A, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset A. The sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3 ’A of a sense strand of one of the sequences of subset A.
  • Any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of the sequences of subset A.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset B.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset B.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset B, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset B, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset B.
  • the sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3 ’A of a sense strand of one of the sequences of subset B. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of the sequences of subset B.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset C.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset C.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset C, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset C, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset C.
  • the sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3 ’A of a sense strand of one of the sequences of subset C. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of the sequences of subset C.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset D.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset D.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset D, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset D, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset D.
  • the sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3 ’A of a sense strand of one of the sequences of subset D. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of the sequences of subset D.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset E.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset E.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset E, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset E, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset E.
  • the sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3 ’A of a sense strand of one of the sequences of subset E. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of the sequences of subset E.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence of an siRNA of subset F.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence of subset F.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset F, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand of subset F, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence of subset F.
  • the sense strand or antisense strand may comprise any modifications described herein.
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3 ’A of a sense strand of one of the sequences of subset F. Any one of the aforementioned siRNA may include an antisense strand that lacks a 5’ U of an antisense strand sequence of any one of the sequences of subset F.
  • the siRNA comprises a sense strand having a sequence in accordance with SEQ ID NO: 7787.
  • the sense strand sequence comprises or consists of sequence at least 75% identical to SEQ ID NO: 7787, at least 80% identical to SEQ ID NO: 7787, at least 85% identical to SEQ ID NO: 7787, at least 90% identical to SEQ ID NO: 7787, or at least 95% identical to SEQ ID NO: 7787.
  • the sense strand sequence comprises or consists of the sequence of SEQ ID NO 7787, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand sequence comprises or consists of the sequence of SEQ ID NO: 7787, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 7787.
  • the sense strand may comprise any modifications or modification pattern described herein.
  • the sense strand may comprise a moiety such as a GalNAc moiety or a lipid moiety.
  • the siRNA comprises an antisense strand having a sequence in accordance with SEQ ID NO: 7839.
  • the antisense strand sequence comprises or consists of sequence at least 75% identical to SEQ ID NO: 7839, at least 80% identical to SEQ ID NO: 7839, at least 85% identical to SEQ ID NO: 7839, at least 90% identical to SEQ ID NO: 7839, or at least 95% identical to SEQ ID NO: 7839.
  • the antisense strand sequence comprises or consists of the sequence of SEQ ID NO 7839, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions.
  • the antisense strand sequence comprises or consists of the sequence of SEQ ID NO: 7839, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 7839.
  • the antisense strand may comprise any modifications or modification pattern described herein.
  • the antisense strand may comprise a moiety such as a GalNAc moiety or a lipid moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of SEQ ID NO: 7787. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5 ’ U of an antisense strand sequence of one of SEQ ID NO: 7839.
  • the siRNA comprises a sense strand having a sequence in accordance with SEQ ID NO: 7799.
  • the sense strand sequence comprises or consists of sequence at least 75% identical to SEQ ID NO: 7799, at least 80% identical to SEQ ID NO: 7799, at least 85% identical to SEQ ID NO: 7799, at least 90% identical to SEQ ID NO: 7799, or at least 95% identical to SEQ ID NO: 7799.
  • the sense strand sequence comprises or consists of the sequence of SEQ ID NO 7799, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand sequence comprises or consists of the sequence of SEQ ID NO: 7799, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 7799.
  • the sense strand may comprise any modifications or modification pattern described herein.
  • the sense strand may comprise a moiety such as a GalNAc moiety or a lipid moiety.
  • the siRNA comprises an antisense strand having a sequence in accordance with SEQ ID NO: 7851.
  • the antisense strand sequence comprises or consists of sequence at least 75% identical to SEQ ID NO: 7851, at least 80% identical to SEQ ID NO: 7851, at least 85% identical to SEQ ID NO: 7851, at least 90% identical to SEQ ID NO: 7851, or at least 95% identical to SEQ ID NO: 7851.
  • the antisense strand sequence comprises or consists of the sequence of SEQ ID NO 7851, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions.
  • the antisense strand sequence comprises or consists of the sequence of SEQ ID NO: 7851, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 7851.
  • the antisense strand may comprise any modifications or modification pattern described herein.
  • the antisense strand may comprise a moiety such as a GalNAc moiety or a lipid moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 7799. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5 ’ U of an antisense strand sequence of one of SEQ ID NO: 7851.
  • the siRNA comprises a sense strand having a sequence in accordance with SEQ ID NO: 7804.
  • the sense strand sequence comprises or consists of sequence at least 75% identical to SEQ ID NO: 7804, at least 80% identical to SEQ ID NO: 7804, at least 85% identical to SEQ ID NO: 7804, at least 90% identical to SEQ ID NO: 7804, or at least 95% identical to SEQ ID NO: 7804.
  • the sense strand sequence comprises or consists of the sequence of SEQ ID NO 7804, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand sequence comprises or consists of the sequence of SEQ ID NO: 7804, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 7804.
  • the sense strand may comprise any modifications or modification pattern described herein.
  • the sense strand may comprise a moiety such as a GalNAc moiety or a lipid moiety.
  • the siRNA comprises an antisense strand having a sequence in accordance with SEQ ID NO: 7856.
  • the antisense strand sequence comprises or consists of sequence at least 75% identical to SEQ ID NO: 7856, at least 80% identical to SEQ ID NO: 7856, at least 85% identical to SEQ ID NO: 7856, at least 90% identical to SEQ ID NO: 7856, or at least 95% identical to SEQ ID NO: 7856.
  • the antisense strand sequence comprises or consists of the sequence of SEQ ID NO 7856, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions.
  • the antisense strand sequence comprises or consists of the sequence of SEQ ID NO: 7856, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID NO: 7856.
  • the antisense strand may comprise any modifications or modification pattern described herein.
  • the antisense strand may comprise a moiety such as a GalNAc moiety or a lipid moiety.
  • Any of the aforementioned siRNAs may include a sense strand that lacks a 3’ A of a sense strand sequence of one of SEQ ID NO: 7804. Any of the aforementioned siRNAs may include an antisense sense strand that lacks a 5 ’ U of an antisense strand sequence of one of SEQ ID NO: 7856.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an antisense oligonucleotide (ASO).
  • ASO antisense oligonucleotide
  • the ASO is 12-30 nucleosides in length. In some embodiments, the ASO is 14-30 nucleosides in length. In some embodiments, the ASO is at least about 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleosides in length, or a range defined by any of the two aforementioned numbers. In some embodiments, the ASO is 15-25 nucleosides in length. In some embodiments, the ASO is 20 nucleosides in length.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an ASO about 12-30 nucleosides in length and comprising a nucleoside sequence complementary to about 12-30 contiguous nucleosides of a full-length human DKK2 pre-mRNA sequence; wherein (i) the oligonucleotide comprises a modification comprising a modified nucleoside and/or a modified intemucleoside linkage, and/or (ii) the composition comprises a pharmaceutically acceptable carrier.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an ASO about 12-30 nucleosides in length and comprising a nucleoside sequence complementary to about 12-30 contiguous nucleosides of a full-length human DKK2 mRNA sequence such as SEQ ID NO: 7599; wherein (i) the oligonucleotide comprises a modification comprising a modified nucleoside and/or a modified intemucleoside linkage, and/or (ii) the composition comprises a pharmaceutically acceptable carrier.
  • the ASO is at least about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more contiguous nucleosides of SEQ ID NO: 7599.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a modification comprising a modified nucleoside and/or a modified intemucleoside linkage, and/or (ii) the composition comprises a pharmaceutically acceptable carrier.
  • the oligonucleotide comprises a modification comprising a modified nucleoside and/or a modified intemucleoside linkage.
  • the oligonucleotide comprises a modified intemucleoside linkage.
  • the modified intemucleoside linkage comprises alkylphosphonate, phosphorothioate, methylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, or carboxymethyl ester, or a combination thereof.
  • the modified intemucleoside linkage comprises one or more phosphorothioate linkages
  • a phosphorothioate may include a nonbridging oxygen atom in a phosphate backbone of the oligonucleotide that is replaced by sulfur.
  • Modified intemucleoside linkages may be included in siRNAs or ASOs. Benefits of the modified intemucleoside linkage may include decreased toxicity or improved pharmacokinetics.
  • the oligonucleotide comprises a duplex consisting of 21-36 nucleotide single strands with base pairing between 17-25 of the base pairs.
  • the duplex comprises blunt-ends at the 5 ’or 3’ ends of each strand.
  • One strand (antisense strand) is complementary to a target mRNA.
  • Each end of the antisense strand has one to five phosphorothioate bonds.
  • the 5’ end has an optional phosphate mimic such as a vinyl phosphonate.
  • the oligonucleotide is used to knock down a target mRNA or a target protein.
  • the sense strand has the same sequence as the target mRNA.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a modified intemucleoside linkage, wherein the oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 modified intemucleoside linkages, or a range of modified intemucleoside linkages defined by any two of the aforementioned numbers. In some embodiments, the oligonucleotide comprises no more than 18 modified intemucleoside linkages. In some embodiments, the oligonucleotide comprises no more than 20 modified intemucleoside linkages.
  • the oligonucleotide comprises 2 or more modified intemucleoside linkages, 3 or more modified intemucleoside linkages, 4 or more modified intemucleoside linkages, 5 or more modified intemucleoside linkages, 6 or more modified intemucleoside linkages, 7 or more modified intemucleoside linkages, 8 or more modified intemucleoside linkages, 9 or more modified intemucleoside linkages, 10 or more modified intemucleoside linkages, 11 or more modified intemucleoside linkages, 12 or more modified intemucleoside linkages, 13 or more modified intemucleoside linkages, 14 or more modified intemucleoside linkages, 15 or more modified intemucleoside linkages, 16 or more modified intemucleoside linkages, 17 or more modified intemucleoside linkages, 18 or more modified intemucleoside linkages, 19 or more modified intemucleoside linkages, or 20 or more modified in
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises the modified nucleoside.
  • the modified nucleoside comprises a locked nucleic acid (LNA), hexitol nucleic acid (HNA), cyclohexene nucleic acid (CeNA), 2'-O-methoxyethyl, 2'-O-alkyl, 2'-O-allyl, 2'-fluoro, or 2'-deoxy, or a combination thereof.
  • the modified nucleoside comprises a LNA.
  • the modified nucleoside comprises a 2’, 4’ constrained ethyl nucleic acid. In some embodiments, the modified nucleoside comprises HLA. In some embodiments, the modified nucleoside comprises CeNA. In some embodiments, the modified nucleoside comprises a 2'-O-methoxyethyl group. In some embodiments, the modified nucleoside comprises a 2'-O-alkyl group. In some embodiments, the modified nucleoside comprises 2’-O-methoxyethyl. In some embodiments, the modified nucleoside comprises a methoxyethyl.
  • position 4 of the sense strand may comprise a methoxyethyl nucleoside such as a 2’-O- methoxyethyl thymine.
  • the modified nucleoside comprises 2'-O-methyl.
  • the modified nucleoside comprises a 2'-O-allyl group.
  • the modified nucleoside comprises a 2'-fluoro group.
  • the modified nucleoside comprises a 2'- deoxy group.
  • the modified nucleoside comprises a 2'-O-methyl nucleoside, 2'- deoxyfluoro nucleoside, 2'-O-N-methylacetamido (2'-0-NMA) nucleoside, a 2'-O- dimethylaminoethoxyethyl (2'-0-DMAE0E) nucleoside, 2'-O-aminopropyl (2'-O-AP) nucleoside, or 2'- ara-F, or a combination thereof.
  • the modified nucleoside comprises a 2'-O-methyl nucleoside.
  • the modified nucleoside comprises a 2'-deoxyfluoro nucleoside.
  • the modified nucleoside comprises a 2'-0-NMA nucleoside. In some embodiments, the modified nucleoside comprises a 2'-0-DMAE0E nucleoside. In some embodiments, the modified nucleoside comprises a 2'-O-aminopropyl (2'-O-AP) nucleoside. In some embodiments, the modified nucleoside comprises 2'-ara-F. In some embodiments, the modified nucleoside comprises one or more 2’- fluoro modified nucleosides. In some embodiments, the modified nucleoside comprises a 2'-O-alkyl modified nucleoside.
  • the modified nucleoside comprises a 2’-O-methyl inosine nucleoside. In some embodiments, the modified nucleoside comprises an acyclic nucleic acid. In some embodiments, the acyclic nucleic is a glycol nucleic acid. In some embodiments, the modified nucleoside comprises an unlocked nucleic acid. Benefits of the modified nucleoside may include decreased toxicity or improved pharmacokinetics.
  • the modified nucleoside comprises a glycol nucleic acid (GNA).
  • GNA glycol nucleic acid
  • the modified nucleoside comprises an unlocked nucleic acid.
  • An unlocked nucleic acid may comprise the following structure:
  • the oligonucleotide comprises a modified nucleoside.
  • the modified nucleoside comprises a locked nucleic acid and an abasic site: are independently an H or a 3’ or 5’ linkage to a nucleotide via a phosphodiester or phosphorothioate bond.
  • the oligonucleotide comprises a phosphate mimic.
  • the phosphate mimic comprises methylphosphonate.
  • An example of a nucleotide that comprises a methylphosphonate is shown below: ’ methylphosphonate 2’-O-Methyl Uridine).
  • the oligonucleotide comprises a duplex consisting of 21-36 nucleotide single strands with base pairing between 17-25 of the base pairs.
  • the duplex comprises blunt-ends at the 5 ’or 3’ ends of each strand.
  • One strand (antisense strand) is complementary to a target mRNA.
  • Each end of the antisense strand has one to five phosphorothioate bonds.
  • the 5’ end has an optional phosphate mimic such as a vinyl phosphonate.
  • the oligonucleotide is used to knock down a target mRNA or a target protein.
  • the sense strand has the same sequence as the target mRNA.
  • the oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 modified nucleosides, or a range of nucleosides defined by any two of the aforementioned numbers. In some embodiments, the oligonucleotide comprises no more than 19 modified nucleosides. In some embodiments, the oligonucleotide comprises no more than 21 modified nucleosides.
  • the oligonucleotide comprises 2 or more modified nucleosides, 3 or more modified nucleosides, 4 or more modified nucleosides, 5 or more modified nucleosides, 6 or more modified nucleosides, 7 or more modified nucleosides, 8 or more modified nucleosides, 9 or more modified nucleosides, 10 or more modified nucleosides, 11 or more modified nucleosides, 12 or more modified nucleosides, 13 or more modified nucleosides, 14 or more modified nucleosides, 15 or more modified nucleosides, 16 or more modified nucleosides, 17 or more modified nucleosides, 18 or more modified nucleosides, 19 or more modified nucleosides, 20 or more modified nucleosides, or 21 or more modified nucleosides.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a moiety attached at a 3 ’ or 5 ’ terminus of the oligonucleotide.
  • moieties include a hydrophobic moiety or a sugar moiety, or a combination thereof.
  • the oligonucleotide is an siRNA having a sense strand, and the moiety is attached to a 5’ end of the sense strand.
  • the oligonucleotide is an siRNA having a sense strand, and the moiety is attached to a 3’ end of the sense strand.
  • the oligonucleotide is an siRNA having an antisense strand, and the moiety is attached to a 5 ’ end of the antisense strand. In some embodiments, the oligonucleotide is an siRNA having an antisense strand, and the moiety is attached to a 3’ end of the antisense strand. In some embodiments, the oligonucleotide is an ASO, and the moiety is attached to a 5’ end of the ASO. In some embodiments, the oligonucleotide is an ASO, and the moiety is attached to a 3’ end of the ASO.
  • the sense strand comprises at least three modified nucleosides, wherein the three modifications comprise a 2’-fluoro modified nucleoside, a 2’-O-methyl modified nucleoside, and 2’- O-methoxyethyl. In some embodiments, the sense strand comprises at least two modified nucleosides, wherein the two modifications comprise a 2’ -fluoro modified nucleoside, a 2’-O-methyl modified nucleoside, and 2’-O-methoxyethyl.
  • each nucleoside of the sense strand comprises a modified nucleoside, wherein the modified nucleosides are selected from the group consisting of a 2’- fluoro modified nucleoside, a 2’-O-methyl modified nucleoside, and 2’-O-methoxyethyl.
  • the sense strand comprises at least a 2’ -fluoro modified nucleoside, a 2’-O-methyl modified nucleoside, and 2’-O-methoxyethyl.
  • the antisense strand is combination of 2’-fluoro and 2’-O-Methyl modifications.
  • each nucleoside of the antisense strand comprises a modified nucleoside, wherein the modified nucleosides are selected from the group consisting of a 2 ’-fluoro modified nucleoside and a 2’-O-methyl modified nucleoside.
  • the sense strand comprises at least a 2 ’-fluoro modified nucleoside and a 2’-O-methyl modified nucleoside.
  • the oligonucleotide may include purines.
  • purines include adenine (A), inosine (I), guanine (G), or modified versions thereof.
  • the oligonucleotide may include pyrimidines. Examples of pyrimidines include cytosine (C), thymine (T), or uracil (U), or modified versions thereof.
  • the sense strand comprises purines and pyrimidines.
  • all purine nucleosides comprise 2’ -fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’-O-methyl and 2’-O-methoxyethyl.
  • all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’ -fluoro and 2’-O-methoxyethyl.
  • all purine nucleosides comprise 2’-O-methoxyethyl, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl. In some embodiments, all pyrimidine nucleosides comprise 2’-fluoro, and all purine nucleosides are modified with a mixture of 2’-O-methyl and 2’-O- methoxyethyl. In some embodiments, all pyrimidine nucleosides comprise 2’-O-methyl, and all purine nucleosides are modified with a mixture of 2 ’-fluoro and 2’-O-methoxyethyl.
  • all pyrimidine nucleosides comprise 2’-O-methoxyethyl, and all purine nucleosides are modified with a mixture of 2 ’-fluoro and 2’-O-methyl.
  • the sense strand may include a 2 ’-deoxy nucleoside.
  • At least one nucleotide at position 4 or 5 of the sense strand comprises a 2’- O-methoxyethyl modified nucleoside. In some embodiments, at least one nucleotide of the sense strand from position 6 to 9 comprise a 2’-fluoro-modified nucleoside. In some embodiments, at least two nucleotides of the sense strand at position 6 to 9 comprise a 2’-fluoro-modified nucleoside. In some embodiments, at least three nucleotides of the sense strand at positions 6 to 9 comprise a 2’ -fluoromodified nucleoside.
  • each nucleotide from positions 6 to 9 of the sense strand comprise a 2’-fluoro-modified nucleoside.
  • at least one nucleotide at position 16 to 20 of the sense strand comprises a 2’-O-methyl modified nucleoside.
  • at least two nucleotides at position 16 to 20 of the sense strand comprise a 2’-O-methyl modified nucleoside.
  • at least three nucleotides at position 16 to 20 of the sense strand comprise a 2’-O-methyl modified nucleoside.
  • at least four nucleotides at position 16 to 20 of the sense strand comprise a 2’-O-methyl modified nucleoside.
  • all nucleotides at position 16 to 20 of the sense strand comprise a 2’-O-methyl modified nucleoside.
  • any of the following is true with regards to the antisense strand: all purine nucleosides comprise 2’-fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl; all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl; all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides comprise 2’-fluoro; all pyrimidine nucleosides comprise 2’-fluoro, and all purine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl; all pyrimidine nucleosides comprise 2’-O-methyl, and all purine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl; all pyrimidine nucleosides comprise
  • all purine nucleosides comprise 2 ’-fluoro, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl. In some embodiments, all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides are modified with a mixture of 2’-fluoro and 2’-O- methyl; all purine nucleosides comprise 2’-O-methyl, and all pyrimidine nucleosides comprise 2’-fluoro.
  • all pyrimidine nucleosides comprise 2’-fluoro, and all purine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl; all pyrimidine nucleosides comprise 2’-O-methyl, and all purine nucleosides are modified with a mixture of 2’-fluoro and 2’-O-methyl. In some embodiments, all pyrimidine nucleosides comprise 2’-O-methyl, and all purine nucleosides comprise 2’- fluoro.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a hydrophobic moiety.
  • the hydrophobic moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide.
  • the hydrophobic moiety may include a lipid such as a fatty acid.
  • the hydrophobic moiety may include a hydrocarbon.
  • the hydrocarbon may be linear.
  • the hydrocarbon may be non-linear.
  • the hydrophobic moiety may include a lipid moiety or a cholesterol moiety, or a combination thereof.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a lipid attached at a 3 ’ or 5 ’ terminus of the oligonucleotide.
  • the lipid comprises cholesterol, myristoyl, palmitoyl, stearoyl, lithocholoyl, docosanoyl, docosahexaenoyl, myristyl, palmityl stearyl, or a-tocopherol, or a combination thereof.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a sugar moiety.
  • the sugar moiety may include an N- acetyl galactose moiety (e.g., an N-acetylgalactosamine (GalNAc) moiety), an N-acetyl glucose moiety (e.g., an N-acetylglucosamine (GlcNAc) moiety), a fucose moiety, or a mannose moiety.
  • the sugar moiety may include 1, 2, 3, or more sugar molecules.
  • the sugar moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide.
  • the sugar moiety may include an N-acetyl galactose moiety.
  • the sugar moiety may include an N-acetylgalactosamine (GalNAc) moiety.
  • the sugar moiety may include an N- acetyl glucose moiety.
  • the sugar moiety may include N-acetylglucosamine (GlcNAc) moiety.
  • the sugar moiety may include a fucose moiety.
  • the sugar moiety may include a mannose moiety. N-acetyl glucose, GlcNAc, fucose, or mannose may be useful for targeting macrophages since they may target or bind a mannose receptor such as CD206.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an N-acetylgalactosamine (GalNAc) moiety.
  • GalNAc may be useful for hepatocyte targeting.
  • the GalNAc moiety may include a bivalent or trivalent branched linker.
  • the oligo may be attached to 1, 2 or 3 GalNAcs through a bivalent or trivalent branched linker.
  • the GalNAc moiety may include 1, 2, 3, or more GalNAc molecules.
  • the GalNAc moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide.
  • the oligonucleotide may include purines.
  • purines include adenine (A), inosine (I), guanine (G), or modified versions thereof.
  • the oligonucleotide may include pyrimidines. Examples of pyrimidines include cytosine (C), thymine (T), or uracil (U), or modified versions thereof.
  • purines of the oligonucleotide comprise 2’-fluoro modified purines. In some embodiments, purines of the oligonucleotide comprise 2’-O-methyl modified purines. In some embodiments, purines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines. In some embodiments, all purines of the oligonucleotide comprise 2’-fluoro modified purines. In some embodiments, all purines of the oligonucleotide comprise 2’-O-methyl modified purines.
  • all purines of the oligonucleotide comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified purines.
  • 2’-O-methyl may include 2’-O-methyl. Where 2’-O-methyl modifications are described, it is contemplated that a 2’-methyl modification may be included, and vice versa.
  • pyrimidines of the oligonucleotide comprise 2 ’-fluoro modified pyrimidines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, pyrimidines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-0-methyl modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’-fluoro modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the oligonucleotide comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified pyrimidines.
  • purines of the oligonucleotide comprise 2’-fluoro modified purines, and pyrimidines of the oligonucleotide comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, purines of the oligonucleotide comprise 2’-O-methyl modified purines, and pyrimidines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines.
  • purines of the oligonucleotide comprise 2’-fluoro modified purines, and pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, purines of the oligonucleotide comprise 2’-O-methyl modified purines, and pyrimidines of the oligonucleotide comprise 2 ’-fluoro modified pyrimidines. In some embodiments, pyrimidines of the oligonucleotide comprise 2’ -fluoro modified pyrimidines, and purines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines.
  • pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines, and purines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines.
  • pyrimidines of the oligonucleotide comprise 2’-fluoro modified pyrimidines, and purines of the oligonucleotide comprise 2’- O-methyl modified purines.
  • pyrimidines of the oligonucleotide comprise 2’-O- methyl modified pyrimidines, and purines of the oligonucleotide comprise 2’-fluoro modified purines.
  • all purines of the oligonucleotide comprise 2 ’-fluoro modified purines, and all pyrimidines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the oligonucleotide comprise 2’-O-methyl modified purines, and all pyrimidines of the oligonucleotide comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified pyrimidines.
  • all purines of the oligonucleotide comprise 2 ’-fluoro modified purines, and all pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the oligonucleotide comprise 2’-O-methyl modified purines, and all pyrimidines of the oligonucleotide comprise 2 ’-fluoro modified pyrimidines.
  • all pyrimidines of the oligonucleotide comprise 2 ’-fluoro modified pyrimidines, and all purines of the oligonucleotide comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines, and all purines of the oligonucleotide comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines.
  • all pyrimidines of the oligonucleotide comprise 2 ’-fluoro modified pyrimidines, and all purines of the oligonucleotide comprise 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the oligonucleotide comprise 2’-O-methyl modified pyrimidines, and all purines of the oligonucleotide comprise 2’ -fluoro modified purines.
  • the oligonucleotide comprises a particular modification pattern.
  • position 9 counting from the 5’ end of the of a strand of the oligonucleotide may have a 2’F modification.
  • position 9 of a strand of the oligonucleotide is a pyrimidine
  • all purines in a strand of the oligonucleotide have a 2’0Me modification.
  • position 9 is the only pyrimidine between positions 5 and 11 of the sense stand, then position 9 is the only position with a 2’F modification in a strand of the oligonucleotide.
  • both of these pyrimidines are the only two positions with a 2’F modification in a strand of the oligonucleotide.
  • position 9 and only two other bases between positions 5 and 11 of a strand of the oligonucleotide are pyrimidines, and those two other pyrimidines are in adjacent positions so that there would be not three 2’F modifications in a row, then any combination of 2’F modifications can be made that give three 2’F modifications in total.
  • a strand of the oligonucleotide of any of the siRNAs comprises a modification pattern which conforms to any or all of these a strand of the oligonucleotide rules.
  • position 9 of a strand of the oligonucleotide when position 9 of a strand of the oligonucleotide is a purine, then all purines in a strand of the oligonucleotide have a 2’0Me modification. In some embodiments, when position 9 is the only purine between positions 5 and 11 of the sense stand, then position 9 is the only position with a 2’F modification in a strand of the oligonucleotide. In some embodiments, when position 9 and only one other base between positions 5 and 11 of a strand of the oligonucleotide are purines, then both of these purines are the only two positions with a 2’F modification in a strand of the oligonucleotide.
  • a strand of the oligonucleotide of any of the siRNAs comprises a modification pattern which conforms to any or all of these a strand of the oligonucleotide rules.
  • position 9 of a strand of the oligonucleotide can be a 2’deoxy.
  • 2’F and 2’0Me modifications may occur at the other positions of a strand of the oligonucleotide.
  • a strand of the oligonucleotide of any of the siRNAs comprises a modification pattern which conforms to these a strand of the oligonucleotide rules.
  • position nine of the sense strand comprises a 2’-fluoro-modified pyrimidine.
  • all purines of the sense strand comprise 2’-O-methyl modified purines.
  • 1, 2, 3, 4, or 5 pyrimidines between positions 5 and 11 comprise a 2 ’fluoromodified pyrimidine, provided there are not three 2’-fluoro-modified pyrimidines in a row.
  • the odd-numbered positions of the antisense strand comprise 2’-O-methyl modified nucleotides.
  • the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides and unmodified deoxyribonucleotide. In some embodiments, the even- numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides, 2’-O-methyl modified nucleotides and unmodified deoxyribonucleotide.
  • position nine of the sense strand comprises a 2’-fluoro-modified pyrimidine; all purines of the sense strand comprises 2’-O-methyl modified purines; 1, 2, 3, 4, or 5 pyrimidines between positions 5 and 11 comprise a 2’fluoro-modified pyrimidine, provided there are not three 2’-fluoro-modified pyrimidines in a row; the odd-numbered positions of the antisense strand comprise 2’-O-methyl modified nucleotides; and the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides and unmodified deoxyribonucleotides.
  • position nine of the sense strand comprises a 2’-fluoro-modified purine.
  • all pyrimidines of the sense strand comprise 2’-O-methyl modified purines.
  • 1, 2, 3, 4, or 5 purines between positions 5 and 11 comprise a 2’fluoro-modified purine, provided there are not three 2’-fluoro-modified purine in a row.
  • the odd-numbered positions of the antisense strand comprise 2’-O-methyl modified nucleotides.
  • the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides and unmodified deoxyribonucleotide.
  • the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides, 2’-O-methyl modified nucleotides and unmodified deoxyribonucleotide.
  • position nine of the sense strand comprises a 2 ’-fluoromodified purine; all pyrimidine of the sense strand comprises 2’-O-methyl modified pyrimidines; 1, 2, 3, 4, or 5 purines between positions 5 and 11 comprise a 2’fluoro-modified purines, provided there are not three 2’-fluoro-modified purines in a row; the odd-numbered positions of the antisense strand comprise 2’-O-methyl modified nucleotides; and the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides and unmodified deoxyribonucleotides. In some embodiments, there are not three 2’-fluoro-modified purines in a row. In some embodiments, there are not three 2’-flu
  • position nine of the sense strand comprises an unmodified deoxyribonucleotide.
  • positions 5, 7, and 8 of the sense strand comprise 2’-fluoro- modifed nucleotides.
  • all pyrimidines in positions 10 to 21 of the sense strand comprise 2’-O-methyl modified pyrimidines and all purines in positions 10 to 21 of the comprise 2’-O- methyl modified purines or 2’-fIuoro-modified purines.
  • the odd-numbered positions of the antisense strand comprise 2’-O-methyl modified nucleotides.
  • the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides and unmodified deoxyribonucleotides. In some embodiments, the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides, 2’-O-methyl modified nucleotides and unmodified deoxyribonucleotides.
  • position nine of the sense strand comprises an unmodified deoxyribonucleotide; positions 5, 7, and 8 of the sense strand comprise 2’-fluoro-modifed nucleotides; all pyrimidines in positions 10 to 21 of the sense strand comprise 2’-O-methyl modified pyrimidines and all purines in positions 10 to 21 of the comprise 2’-O-methyl modified purines or 2’-fluoro-modified purines; the odd-numbered positions of the antisense strand comprise 2’-O-methyl modified nucleotides; and the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides and unmodified deoxyribonucleotides.
  • position nine of the sense strand comprises an unmodified deoxyribonucleotide.
  • positions 5, 7, and 8 of the sense strand comprise 2’-fluoro- modifed nucleotides.
  • all purines in positions 10 to 21 of the sense strand comprise 2’-O-methyl modified purines and all pyrimidines in positions 10 to 21 of the comprise 2’-O-methyl modified pyrimidines or 2’-fluoro-modified pyrimidines.
  • the odd-numbered positions of the antisense strand comprise 2’-O-methyl modified nucleotides.
  • the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides and unmodified deoxyribonucleotides. In some embodiments, the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides, 2’-O-methyl modified nucleotides and unmodified deoxyribonucleotides.
  • position nine of the sense strand comprises an unmodified deoxyribonucleotide; positions 5, 7, and 8 of the sense strand comprise 2’-fluoro-modifed nucleotides; all purines in positions 10 to 21 of the sense strand comprise 2’-O-methyl modified purines and all pyrimidines in positions 10 to 21 of the comprise 2’-O-methyl modified pyrimidines or 2’-fluoro- modified pyrimidines; the odd-numbered positions of the antisense strand comprise 2’-O-methyl modified nucleotides; and the even-numbered positions of the antisense strand comprise 2’fluoro-modified nucleotides and unmodified deoxyribonucleotide.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a targeting ligand.
  • the moiety includes a negatively charged group attached at a 5’ end of the oligonucleotide. This may be referred to as a 5 ’-end group.
  • the negatively charged group is attached at a 5’ end of an antisense strand of an siRNA disclosed herein.
  • the 5 ’-end group may be or include a 5’-end phosphorothioate, 5’-end phosphorodithioate, 5’-end vinylphosphonate (5’-VP), 5’- end methylphosphonate, 5’-end cyclopropyl phosphonate, or a 5’-deoxy-5’-C-malonyl.
  • the 5’-end group may comprise 5 ’-VP.
  • the 5 ’-VP comprises a trans-vinylphosphonate or cis- vinylphosphonate.
  • the 5 ’-end group may include an extra 5’ phosphate.
  • a combination of 5 ’-end groups may be used.
  • the oligonucleotide includes a negatively charged group.
  • the negatively charged group may aid in cell or tissue penetration.
  • the negatively charged group may be attached at a 5’ or 3’ end (e.g. a 5’ end) of the oligonucleotide. This may be referred to as an end group.
  • the end group may be or include a phosphorothioate, phosphorodithioate, vinylphosphonate, methylphosphonate, cyclopropyl phosphonate, or a deoxy-C-malonyl.
  • the end group may include an extra 5’ phosphate such as an extra 5’ phosphate.
  • a combination of end groups may be used.
  • the oligonucleotide includes a phosphate mimic.
  • the phosphate mimic comprises vinyl phosphonate.
  • the vinyl phosphonate comprises a trans-vinylphosphonate.
  • the vinyl phosphonate comprises a cis- vinylphosphonate.
  • An example of a nucleotide that includes a vinyl phosphonate is shown below.
  • the vinyl phosphonate increases the stability of the oligonucleotide. In some embodiments, the vinyl phosphonate increases the accumulation of the oligonucleotide in tissues. In some embodiments, the vinyl phosphonate protects the oligonucleotide from an exonuclease or a phosphatase. In some embodiments, the vinyl phosphonate improves the binding affinity of the oligonucleotide with the siRNA processing machinery.
  • the oligonucleotide includes 1 vinyl phosphonate. In some embodiments, the oligonucleotide includes 2 vinyl phosphonates. In some embodiments, the oligonucleotide includes 3 vinyl phosphonates. In some embodiments, the oligonucleotide includes 4 vinyl phosphonates. In some embodiments, the antisense strand of the oligonucleotide comprises a vinyl phosphonate at the 5’ end. In some embodiments, the antisense strand of the oligonucleotide comprises a vinyl phosphonate at the 3’ end.
  • the sense strand of the oligonucleotide comprises a vinyl phosphonate at the 5’ end. In some embodiments, the sense strand of the oligonucleotide comprises a vinyl phosphonate at the 3’ end.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a hydrophobic moiety.
  • An oligonucleotide comprising a hydrophobic moiety may include, or be referred to as a hydrophobic conjugate. Hydrophobic moieties may be useful for enhancing cellular uptake.
  • the hydrophobic moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide.
  • the hydrophobic moiety may include a lipid such as a fatty acid.
  • the hydrophobic moiety may include a hydrocarbon.
  • the hydrocarbon may be linear.
  • the hydrocarbon may be non-linear.
  • the hydrophobic moiety may include a lipid moiety or a cholesterol moiety, or a combination thereof. In some embodiments, the hydrophobic moiety includes a cyclohexanyl. In some embodiments, the hydrophobic moiety includes a lipid. In some embodiments, the hydrophobic moiety is used in a specific format described herein. In some embodiments, the hydrophobic moiety is attached at a 5’ end of a sense strand without any phosphorothioate groups or linkages at the 5’ end. The hydrophobic moiety may include an esterified lipid.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a lipid attached at a 3 ’ or 5 ’ terminus of the oligonucleotide.
  • the lipid comprises cholesterol, myristoyl, palmitoyl, stearoyl, lithocholoyl, docosanoyl, docosahexaenoyl, myristyl, palmityl, stearyl, or a-tocopherol, or a combination thereof.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a hydrophobic ligand or moiety.
  • the hydrophobic ligand or moiety comprises cholesterol.
  • the hydrophobic ligand or moiety comprises a cholesterol derivative.
  • the hydrophobic ligand or moiety is attached at a 3’ terminus of the oligonucleotide.
  • the hydrophobic ligand or moiety s attached at a 5’ terminus of the oligonucleotide.
  • the composition comprises a sense strand, and the hydrophobic ligand or moiety is attached to the sense strand (e.g. attached to a 5’ end of the sense strand, or attached to a 3’ end of the sense strand).
  • the composition comprises an antisense strand, and the hydrophobic ligand or moiety is attached to the antisense strand (e.g. attached to a 5’ end of the antisense strand, or attached to a 3’ end of the antisense strand).
  • the composition comprises a hydrophobic ligand or moiety attached at a 3’ or 5’ terminus of the oligonucleotide.
  • a hydrophobic moiety is attached to the oligonucleotide (e.g. a sense strand and/or an antisense strand of a siRNA). In some embodiments, a hydrophobic moiety is attached at a 3’ terminus of the oligonucleotide. In some embodiments, a hydrophobic moiety is attached at a 5’ terminus of the oligonucleotide. In some embodiments, the hydrophobic moiety comprises cholesterol. In some embodiments, the hydrophobic moiety includes a cyclohexanyl. The hydrophobic moiety may include an esterified lipid.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a lipid attached at a 3 ’ or 5 ’ terminus of the oligonucleotide. In some embodiments, a lipid is attached at a 3’ terminus of the oligonucleotide. In some embodiments, a lipid is attached at a 5’ terminus of the oligonucleotide.
  • the lipid comprises cholesterol, myristoyl, palmitoyl, stearoyl, lithocholoyl, docosanoyl, docosahexaenoyl, myristyl, palmityl, stearyl, or a-tocopherol, or a combination thereof.
  • the lipid comprises stearyl, lithocholyl, docosanyl, docosahexaenyl, or myristyl.
  • the lipid comprises cholesterol.
  • the lipid includes a sterol such as cholesterol.
  • the lipid comprises stearyl, t-butylphenol, n-butylphenol, octylphenol, dodecylphenol, phenyl n-dodecyl, octadecylbenzamide, hexadecylbenzamide, or octadecylcyclohexyl.
  • the lipid comprises phenyl para C12. The lipid moiety may be esterified.
  • the oligonucleotide comprises any aspect of the following structure:
  • the oligonucleotide comprises any aspect of the following structure: some embodiments, the oligonucleotide comprises any aspect of the following structure: some embodiments, the oligonucleotide comprises any aspect of the following structure aspect included in the oligonucleotide may include the entire structure, or may include the lipid moiety, of any of the structures shown.
  • n is 1-3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.
  • R is an alkyl group. In some embodiments, the alkyl group contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbons.
  • the alkyl group contains 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbons, or a range defined by any two of the aforementioned numbers of carbons. In some embodiments, the alkyl group contains 4-18 carbons. In some embodiments, the lipid moiety comprises an alcohol or ether.
  • the lipid includes a fatty acid.
  • the lipid comprises a lipid depicted in Table 1.
  • the example lipid moieties in Table 1 are shown attached at a 5’ end of an oligonucleotide, in which the 5’ terminal phosphate of the oligonucleotide is shown with the lipid moiety.
  • a lipid moiety in Table 1 may be attached at a different point of attachment than shown.
  • the point of attachment of any of the lipid moieties in the table may be at a 3’ oligonucleotide end.
  • the lipid is used for targeting the oligonucleotide to a non- hepatic cell or tissue.
  • the lipid or lipid moiety includes 16 to 18 carbons. In some embodiments, the lipid includes 16 carbons. In some embodiments, the lipid includes 17 carbons. In some embodiments, the lipid includes 18 carbons. In some embodiments, the lipid moiety includes 16 carbons. In some embodiments, the lipid moiety includes 17 carbons. In some embodiments, the lipid moiety includes 18 carbons. In some embodiments, the lipid moiety includes 19 carbons. In some embodiments, the lipid moiety includes 20 carbons.
  • the hydrophobic moiety may include a linker that comprises a carbocycle.
  • the carbocycle may be six-membered. Some examples of a carbocycle include phenyl or cyclohexyl.
  • the linker may include a phenyl.
  • the linker may include a cyclohexyl.
  • the lipid may be attached to the carbocycle, which may in turn be attached at a phosphate (e.g. 5’ or 3’ phosphate) of the oligonucleotide.
  • the lipid or hydrocarbon, and the end of the sense are connected to the phenyl or cyclohexyl linker in the 1,4; 1,3; or 1,2 substitution pattern (e.g. the para, meta, or ortho phenyl configuration).
  • the lipid or hydrocarbon, and the end of the sense are connected to the phenyl or cyclohexyl linker in the
  • the lipid may be attached to the carbocycle in the 1,4 substitution pattern relative to the oligonucleotide.
  • the lipid may be attached to the carbocycle in the 1,3 substitution pattern relative to the oligonucleotide.
  • the lipid may be attached to the carbocycle in the 1,2 substitution pattern relative to the oligonucleotide.
  • the lipid may be attached to the carbocycle in the ortho orientation relative to the oligonucleotide.
  • the lipid may be attached to the carbocycle in the para orientation relative to the oligonucleotide.
  • the lipid may be attached to the carbocycle in the meta orientation relative to the oligonucleotide.
  • the lipid moiety may comprise or consist of the following structure
  • the lipid moiety comprises or consist of the following structure: comprises the following structure: lipid moiety comprises or consist of the following structure: some embodiments, the dotted line indicates a covalent connection.
  • the covalent connection may between an end of the sense or antisense strand. For example, the connection may be to the 5’ end of the sense strand.
  • n is 0-3. In some embodiments, n is 1-3. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • R is an alkyl group.
  • the alkyl group contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbons.
  • the alkyl group contains 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbons, or a range defined by any two of the aforementioned numbers of carbons.
  • R comprises or consists of an alkyl group containing 4-18 carbons.
  • the lipid moiety may be attached at a 5’ end of the oligonucleotide. The 5’ end may have one phosphate linking the lipid moiety to a 5’ carbon of a sugar of the oligonucleotide.
  • the 5’ end may have two phosphates linking the lipid moiety to a 5’ carbon of a sugar of the oligonucleotide.
  • the 5 ’ end may have three phosphates linking the lipid moiety to a 5 ’ carbon of a sugar of the oligonucleotide.
  • the 5’ end may have one phosphate connected to the 5’ carbon of a sugar of the oligonucleotide, where the one phosphate is connected to the lipid moiety.
  • the 5’ end may have two phosphates connected to the 5 ’ carbon of a sugar of the oligonucleotide, where the one of the two phosphates is connected to the lipid moiety.
  • the 5’ end may have three phosphates connected to the 5’ carbon of a sugar of the oligonucleotide, where the one of the three phosphates is connected to the lipid moiety.
  • the sugar may include a ribose.
  • the sugar may include a deoxyribose.
  • the sugar may be modified a such as a 2’ modified sugar (e.g. a 2’-0-methyl or 2’-fluoro ribose).
  • a phosphate of the 5’ end may include a modification such as a sulfur in place of an oxygen.
  • Two phosphates of the 5’ end may include a modification such as a sulfur in place of an oxygen.
  • Three phosphates of the 5’ end may include a modification such as a sulfur in place of an oxygen.
  • the oligonucleotide includes 1 lipid moiety. In some embodiments, the oligonucleotide includes 2 lipid moieties. In some embodiments, the oligonucleotide includes 3 lipid moieties. In some embodiments, the oligonucleotide includes 4 lipid moieties.
  • Some embodiments relate to a method of making an oligonucleotide comprising a hydrophobic conjugate.
  • a strategy for making hydrophobic conjugates may include use of a phosphoramidite reagent based upon a 6-membered ring alcohol such as a phenol or cyclohexanol. The phosphoramidite may be reacted to a nucleotide to connect the nucleotide to the hydrophobic moiety, and thereby produce the hydrophobic conjugate.
  • Some examples of phosphoramidite reagents that may be used to produce a hydrophobic conjugate are provided as follows: . In some embodiments, n is 1 -3. In some embodiments, n is 1. In some embodiments, n is 2.
  • n is 3.
  • R is an alkyl group.
  • the alkyl group contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbons.
  • the alkyl group contains 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbons, or a range defined by any two of the aforementioned numbers of carbons.
  • R comprises or consists of an alkyl group containing 4-18 carbons. Any one of the phosphoramidite reagents may be reacted to a 5’ end of an oligonucleotide to produce an oligonucleotide comprising a hydrophobic moiety.
  • the phosphoramidite reagents is reacted to a 5’ end of a sense strand of an siRNA.
  • the sense strand may then be hybridized to an antisense strand to form a duplex.
  • the hybridization may be performed by incubating the sense and antisense strands in solution at a given temperature.
  • the temperature may be gradually reduced.
  • the temperature may comprise or include a temperature comprising an annealing temperature for the sense and antisense strands.
  • the temperature may be below or include a temperature below the annealing temperature for the sense and antisense strands.
  • the temperature may be below a melting temperature of the sense and antisense strands.
  • the lipid may be attached to the oligonucleotide by a linker.
  • the linker may include a polyethyleneglycol (e.g. tetraethyleneglycol).
  • the modifications described herein may be useful for delivery to a cell or tissue, for example, extrahepatic delivery or targeting of an oligonucleotide composition.
  • the modifications described herein may be useful for targeting an oligonucleotide composition to a cell or tissue.
  • ETL2 may be conjugated to an oligonucleotide using the following reagent: 2.
  • Sugar moieties may be conjugated to an oligonucleotide using the following reagent: 2.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a sugar moiety.
  • the sugar moiety may include an N- acetyl galactose moiety (e.g. an N-acetylgalactosamine (GalNAc) moiety), an N-acetyl glucose moiety (e.g. an N-acetylglucosamine (GlcNAc) moiety), a fucose moiety, or a mannose moiety.
  • the sugar moiety may include 1, 2, 3, or more sugar molecules.
  • the sugar moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide.
  • the sugar moiety may include an N-acetyl galactose moiety.
  • the sugar moiety may include an N-acetylgalactosamine (GalNAc) moiety.
  • the sugar moiety may include an N-acetyl glucose moiety.
  • the sugar moiety may include N-acetylglucosamine (GlcNAc) moiety.
  • the sugar moiety may include a fucose moiety.
  • the sugar moiety may include a mannose moiety. N-acetyl glucose, GlcNAc, fucose, or mannose may be useful for targeting macrophages when they target or bind a mannose receptor such as CD206.
  • the sugar moiety may be useful for binding or targeting an asialoglycoprotein receptor such as an asialoglycoprotein receptor of a hepatocyte.
  • the GalNAc moiety may bind to an asialoglycoprotein receptor.
  • the GalNAc moiety may target a hepatocyte.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an N-acetylgalactosamine (GalNAc) moiety.
  • GalNAc may be useful for hepatocyte targeting.
  • the GalNAc moiety may include a bivalent or trivalent branched linker.
  • the oligo may be attached to 1, 2 or 3 GalNAcs through a bivalent or trivalent branched linker.
  • the GalNAc moiety may include 1, 2, 3, or more GalNAc molecules.
  • the GalNAc moiety may be attached at a 3’ or 5’ terminus of the oligonucleotide.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an N-acetylgalactosamine (GalNAc) ligand for hepatocyte targeting.
  • the composition comprises GalNAc.
  • the composition comprises a GalNAc derivative.
  • the GalNAc ligand is attached at a 3’ terminus of the oligonucleotide.
  • the GalNAc ligand is attached at a 5’ terminus of the oligonucleotide.
  • the composition comprises a sense strand, and the GalNAc ligand is attached to the sense strand (e.g. attached to a 5’ end of the sense strand, or attached to a 3’ end of the sense strand).
  • the composition comprises an antisense strand, and the GalNAc ligand is attached to the antisense strand (e.g. attached to a 5’ end of the antisense strand, or attached to a 3’ end of the antisense strand).
  • the composition comprises a GalNAc ligand attached at a 3’ or 5’ terminus of the oligonucleotide.
  • compositions comprising an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises a GalNAc moiety.
  • the GalNAc moiety may be included in any formula, structure, or GalNAc moiety shown below.
  • described herein is a compound (e.g. oligonucleotide) represented by Formula (I) or (II):
  • Q is selected from:
  • C3-10 carbocycle optionally substituted with one or more substituents independently selected from halogen, -CN, -NO 2 , -OR 7 , -SR 7 , -N(R 7 ) 2 , -C(O)R 7 , -C(O)N(R 7 ) 2 , -N(R 7 )C(O)R 7 - N(R 7 )C(O)N(R 7 ) 2 , -OC(O)N(R 7 ) 2 , -N(R 7 )C(O)OR 7 , -C(O)OR 7 , -OC(O)R 7 , -S(O)R 7 , and Ci 6 alkyl, wherein the C1-6 alkyl, is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO2, and -NH2;
  • R 1 is a linker selected from:
  • C1-6 alkyl optionally substituted with one or more substituents independently selected from halogen, -OR 7 , -SR 7 , -N(R 7 ) 2 , -C(O)R 7 , -C(O)N(R 7 ) 2 , -N(R 7 )C(O)R 7 -N(R 7 )C(O)N(R 7 ) 2 , - OC(O)N(R 7 ) 2 , -N(R 7 )C(O)OR 7 , -C(O)OR 7 , -OC(O)R 7 , and -S(O)R 7 ;
  • R 3 and R 4 are each independently selected from:
  • each R 5 is independently selected from: -OC(O)R 7 , -OC(O)N(R 7 ) 2 , -N(R 7 )C(O)R 7 -N(R 7 )C(O)N(R 7 ) 2 , -N(R 7 )C(O)OR 7 , -C(O)OR 7 , -OC(O)R 7 , and -S(O)R 7 ; each R 5 is independently selected from: -OC(O)R 7 , -OC(O)N(R 7 ) 2 , -N(R 7 )C(O)R 7 -N(R 7 )C(O)N(R 7 ) 2 , -N(R 7 )C(O)OR 7 , -C(O)R 7 , and -C(O)OR 7 ; each R 5 is independently selected from: -OC(O)R 7 , -OC(O)N(R 7 ) 2 ,
  • each w is independently selected from any value from 1 to 10. In some embodiments, each w is independently selected from any value from 1 to 5. In some embodiments, each w is 1. In some embodiments, each v is independently selected from any value from 1 to 10. In some embodiments, each v is independently selected from any value from 1 to 5. In some embodiments, each v is 1. In some embodiments, n is selected from any value from 1 to 10. In some embodiments, n is selected from any value from 1 to 5. In some embodiments, n is 2. In some embodiments, m is selected from any value from 1 to 10. In some embodiments, m is selected from any value from 1 to 5. In some embodiments, m is selected from 1 and 2.
  • z is 3 and Y is C.
  • Q is selected from C5-6 carbocycle optionally substituted with one or more substituents independently selected from halogen, -CN, -NO 2 , -OR 7 , -SR 7 , -N(R 7 ) 2 , -C(O)R 7 , -C(O)N(R 7 ) 2 , -N(R 7 )C(O)R 7 - N(R 7 )C(O)N(R 7 ) 2 , -OC(O)N(R 7 ) 2 , -N(R 7 )C(O)OR 7 , -C(O)OR 7 , -OC(O)R 7 , and -S(O)R 7 .
  • Q is selected from C5-6 carbocycle optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO 2 , and -NH 2 .
  • Q is selected from phenyl and cyclohexyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO 2 , and -NH 2 .
  • Q is selected from phenyl.
  • Q is selected from cyclohexyl.
  • R 1 is selected from -OP(O)(OR 7 )O-, -SP(O)(OR 7 )O-, -OP(S)(OR 7 )O-, -OP(O)(SR 7 )O-, - OP(O)(OR 7 )S-, -OP(O)(O )O-, -SP(O)(O )O-, -OP(S)(O )O-, -OP(O)(S )O-, -OP(O)(O )S-, -OP(O)(OR 7 )NR 7 -, -OP(O)(N(R 7 ) 2 )NR 7 -, -OP(OR 7 )O-, -OP(N(R 7 ) 2 )O-, -OP(OR 7 )N(R 7 )-, and -OPN(R 7 ) 2 .
  • R 1 is selected from -OP(O)(OR 7 )O-, -SP(O)(OR 7 )O-, -OP(S)(OR 7 )O-, - OP(O)(SR 7 )O-, -OP(O)(OR 7 )S-, -OP(O)(O )O-, -SP(O)(O )O-, -OP(S)(O )O-, -OP(O)(S )O-, -OP(O)(S )O-, -OP(O)(O )S-, and -OP(OR 7 )O-.
  • R 1 is selected from -OP(O)(OR 7 )O-, -OP(S)(OR 7 )O-, - OP(O)(O )O-, -OP(S)(O )O-, -OP(O)(S )O-, and -OP(OR 7 )O-. In some embodiments, R 1 is selected from - OP(O)(OR 7 )O- and -OP(OR 7 )O-.
  • R 2 is selected from C1-3 alkyl substituted with one or more substituents independently selected from halogen, -OR 7 , -OC(O)R 7 , -SR 7 , -N(R 7 )2, -C(O)R 7 , and -S(O)R 7 .
  • R 2 is selected from C1-3 alkyl substituted with one or more substituents independently selected from -OR 7 , -OC(O)R 7 , -SR 7 , and -N(R 7 )2.
  • R 2 is selected from C1-3 alkyl substituted with one or more substituents independently selected from -OR 7 and - OC(O)R 7 .
  • R 3 is selected from halogen, -OR 7 , -SR 7 , -N(R 7 )2, -C(O)R 7 , -OC(O)R 7 , and -S(O)R 7 In some embodiments, R 3 is selected from -OR 7 -SR 7 , -OC(O)R 7 , and -N(R 7 )2. In some embodiments, R 3 is selected from -OR 7 - and -OC(O)R 7 .
  • R 4 is selected from halogen, -OR 7 , -SR 7 , -N(R 7 )2, -C(O)R 7 , -OC(O)R 7 , and -S(O)R 7 In some embodiments, R 4 is selected from -OR 7 -SR 7 , -OC(O)R 7 , and -N(R 7 )2 In some embodiments, R 4 is selected from -OR 7 - and -OC(O)R 7 .
  • R 5 is selected from -OC(O)R 7 , -OC(O)N(R 7 )2, -N(R 7 )C(O)R 7 -N(R 7 )C(O)N(R 7 )2, and -N(R 7 )C(O)OR 7 . In some embodiments, R 5 is selected from -OC(O)R 7 and -N(R 7 )C(O)R 7 .
  • each R 7 is independently selected from C1-6 alkyl optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, and -SH.
  • Q is phenyl or cyclohexyl, each of which is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO2, -NH2, and C1-3 alkyl;
  • R 1 is selected from -OP(O)(OR 7 )O-, -OP(S)(OR 7 )O-, -0P(0)(0 )0-, -OP(S)(O )0-, -OP(O)(S )0-, and - OP(OR 7 )O-;
  • R 2 is Ci alkyl substituted with -OH or -0C(0)CH3;
  • R 3 is -OH or -0C( some embodiments, the compound compri
  • the oligonucleotide (J) is attached at a 5’ end or a 3’ end of the oligonucleotide.
  • the oligonucleotide comprises DNA.
  • the oligonucleotide comprises RNA.
  • the oligonucleotide comprises one or more modified intemucleoside linkages.
  • the one or more modified intemucleoside linkages comprise alkylphosphonate, phosphorothioate, methylphosphonate, phosphorodithioate, alkylphosphonothioate, phosphoramidate, carbamate, carbonate, phosphate triester, acetamidate, or carboxymethyl ester, or a combination thereof.
  • the oligonucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 modified intemucleoside linkages.
  • the compound binds to an asialoglycoprotein receptor.
  • the compound targets a hepatocyte.
  • Some embodiments include the following, where J is the oligonucleotide:
  • J may include one or more additional phosphates, or one or more phosphorothioates linking to the oligonucleotide.
  • J may include one or more additional phosphates linking to the oligonucleotide.
  • J may include one or more phosphorothioates linking to the oligonucleotide.
  • J is the oligonucleotide: J may include one or more additional phosphates, or one or more phosphorothioates linking to the oligonucleotide. J may include one or more additional phosphates linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide.
  • Some embodiments include the following, where J is the oligonucleotide:
  • J may include one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J may include one or more phosphates linking to the oligonucleotide.
  • J may include a phosphate linking to the oligonucleotide.
  • J may include one or more phosphorothioates linking to the oligonucleotide.
  • J may include a phosphorothioate linking to the oligonucleotide.
  • Some embodiments include the following, where J is the oligonucleotide:
  • J The structure in this compound attached to the oligonucleotide (J) may be referred to as “ETL17”, and is an example of a GalNAc moiety.
  • J may include one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J may include one or more phosphates linking to the oligonucleotide.
  • J may include a phosphate linking to the oligonucleotide.
  • J may include one or more phosphorothioates linking to the oligonucleotide.
  • J may include a phosphorothioate linking to the oligonucleotide.
  • Some embodiments include the following, where the phosphate or “5”’ indicates a connection to the oligonucleotide: [00135] Some embodiments include the following, where the phosphate or “5”’ indicates a connection to
  • Some embodiments include the following, where the phosphate or “5”’ indicates a connection to
  • J is the oligonucleotide: include one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J may include one or more phosphates linking to the oligonucleotide.
  • J may include a phosphate linking to the oligonucleotide.
  • J may include one or more phosphorothioates linking to the oligonucleotide.
  • J may include a phosphorothioate linking to the oligonucleotide.
  • Some embodiments include the following, where J is the oligonucleotide:
  • J The structure in this compound attached to the oligonucleotide (J) may be referred to as “ETL1,” and is an example of a GalNAc moiety.
  • J may include one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J may include one or more phosphates linking to the oligonucleotide.
  • J may include a phosphate linking to the oligonucleotide.
  • J may include one or more phosphorothioates linking to the oligonucleotide.
  • J may include a phosphorothioate linking to the oligonucleotide.
  • J may include one or more additional phosphates linking to the oligonucleotide.
  • J may include one or more phosphorothioates linking to the oligonucleotide.
  • Some embodiments include the following, where J is the oligonucleotide:
  • J may include one or more additional phosphates, or one or more phosphorothioates linking to the oligonucleotide. J may include one or more additional phosphates linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. [00142] Some embodiments include the following, where J is the oligonucleotide: or more phosphates or phosphorothioates linking to the oligonucleotide. J may include one or more phosphates linking to the oligonucleotide. J may include a phosphate linking to the oligonucleotide. J may include one or more phosphorothioates linking to the oligonucleotide. J may include a phosphorothioate linking to the oligonucleotide.
  • Some embodiments include the following, where J is the oligonucleotide:
  • J The structure in this compound attached to the oligonucleotide (J) may be referred to as “ETL17,” and is an example of a GalNAc moiety.
  • J may include one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J may include one or more phosphates linking to the oligonucleotide.
  • J may include a phosphate linking to the oligonucleotide.
  • J may include one or more phosphorothioates linking to the oligonucleotide.
  • J may include a phosphorothioate linking to the oligonucleotide.
  • compositions comprising an oligonucleotide that inhibits the expression of a target gene, wherein the oligonucleotide comprises a GalNAc moiety.
  • the GalNAc moiety may be included in any formula, structure, or GalNAc moiety shown below.
  • described herein is a compound (e.g. oligonucleotide) represented by Formula (III), (IV), or (V):
  • Q is selected from: C3-20 cyclic, heterocyclic or acyclic linker optionally substituted with one or more substituents independently selected from halogen, -CN, -NO2, -OR 7 , -SR 7 , -N(R 7 )2, -C(O)R 7 , - C(O)N(R 7 ) 2 , -N(R 7 )C(O)R 7 -N(R 7 )C(O)N(R 7 ) 2 , -OC(O)N(R 7 ) 2 , -N(R 7 )C(O)OR 7 , -C(O)OR 7 , -OC(O)R 7 , - S(O)R 7 , and Ci-e alkyl, wherein the C1-6 alkyl, is optionally substituted with one or more substituents independently selected from halogen, -CN, -OH, -SH, -NO2, and -NH2;
  • R 1 is a linker selected from: -O-, -S-, -N(R 7 )-, -C(O)-, -C(O)N(R 7 )-, -N(R 7 )C(O)-_ -N(R 7 )C(O)N(R 7 )-, -OC(O)N(R 7 )-, -N(R 7 )C(O)O-, -C(O)O-, -OC(O)-, -S(O)-, -S(O) 2 -, -OS(O) 2 -, -OP(O)(OR 7 )O-, - SP(O)(OR 7 )O-, -OP(S)(OR 7 )O-, -OP(O)(SR 7 )O-, -OP(O)(OR 7 )S-, -OP(O)(O-, -SP(O)(O )O-,
  • sugar moieties comprising the following structure, where J is an oligonucleotide:
  • J The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “L96,” and is an example of a GalNAc moiety.
  • J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J in some instances comprises a phosphate linking to the oligonucleotide.
  • J in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • sugar moieties comprising the following structure, where J is an oligonucleotide:
  • J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J in some instances comprises a phosphate linking to the oligonucleotide.
  • J in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • sugar moieties comprising the following structure, where J is an oligonucleotide:
  • J The structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “GluGalNAc,” and is an example of a GalNAc moiety.
  • J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J in some instances comprises a phosphate linking to the oligonucleotide.
  • J in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • sugar moieties comprising the following structure, where J and K are independently H, a GalNAc moiety or oligonucleotides:
  • the structures in these compounds in some instances are attached to the oligonucleotide (J or K) and referred to as “ademA GalNAc, ademG GalNAc, ademC GalNAc, or ademU GalNAc” depending on the base used in the nucleotide.
  • ademA GalNAc ademG GalNAc
  • ademC GalNAc ademC GalNAc
  • ademU GalNAc oligonucleotide
  • 2-4 GalNAc moieties are attached to the oligonucleotide.
  • J and K may in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J and K in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J and K in some instances comprises a phosphate linking to the oligonucleotide.
  • J and K in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J and K in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • sugar moieties comprising the following structure, where R is an oligonucleotide:
  • R in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • R in some instances comprises one or more phosphates linking to the oligonucleotide.
  • R in some instances comprises a phosphate linking to the oligonucleotide.
  • R in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • R in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • sugar moieties comprising the following structure, where J is an oligonucleotide:
  • J The structure in this compound attached to the oligonucleotide (J) may be referred to as “K2GalNAc,” and is an example of a GalNAc moiety.
  • J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J in some instances comprises a phosphate linking to the oligonucleotide.
  • J in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • J is an oligonucleotide and X is S or O: .
  • the structure in this compound attached to the oligonucleotide (J) in some instances is referred to as “ST23,” and is an example of a GalNAc moiety.
  • J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J in some instances comprises a phosphate linking to the oligonucleotide.
  • J in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • sugar moieties comprising the following structure, where J is an oligonucleotide:
  • J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J in some instances comprises a phosphate linking to the oligonucleotide.
  • J in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • sugar moieties comprising the following structure, where J or K comprises an oligonucleotide:
  • the structures in these compounds in some instances are attached to the oligonucleotide (J or K), referred to as “PyrGalNAc”, “PipGalNAc” and “TEG-GalNAc” are examples of GalNAc moieties.
  • 2-4 GalNAc moieties are attached oligonucleotide.
  • J and K in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J and K in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J and K in some instances comprises a phosphate linking to the oligonucleotide.
  • J and K in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J and K in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • sugar moieties comprising the following structure, where J is an oligonucleotide:
  • J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J in some instances comprises a phosphate linking to the oligonucleotide.
  • J in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • sugar moieties comprising the following structure, where Nu is an oligonucleotide:
  • Nu in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide. Nu in some instances comprises one or more phosphates linking to the oligonucleotide. Nu in some instances comprises a phosphate linking to the oligonucleotide. Nu in some instances comprises one or more phosphorothioates linking to the oligonucleotide. Nu in some instances comprises a phosphorothioate linking to the oligonucleotide. [00164] Provided herein are sugar moieties comprising the following structure, where J is an oligonucleotide:
  • J in some instances comprises one or more phosphates or phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises one or more phosphates linking to the oligonucleotide.
  • J in some instances comprises a phosphate linking to the oligonucleotide.
  • J in some instances comprises one or more phosphorothioates linking to the oligonucleotide.
  • J in some instances comprises a phosphorothioate linking to the oligonucleotide.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2 wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the sense strand comprises modification pattern IS: 5'-NfsnsNfnNfnNfNfNfnNfnNfnNfnNfnNfsnsn-3'.
  • NT is a 2’-fluoro-modified nucleoside
  • n is a 2’-O-methyl modified nucleoside
  • s is a phosphorothioate or phosphate.
  • the sense strand comprises modification pattern 2S: 5’-nsnsnnnNfnNfNfNfnnnnnnnnsn-3’, wherein “Nf ’ is a 2 ’-fluoro -modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, and “s” is a phosphorothioate or phosphate.
  • the sense strand comprises modification pattern 3S: 5’-nsnsnnNfhNfhNfnnnnnnnnsn-3’, wherein “NT’ is a 2’-fluoro-modified nucleoside, “n” is a 2’- O-methyl modified nucleoside, and “s” is a phosphorothioate or phosphate.
  • the sense strand comprises modification pattern 4S: 5'-NfsnsNfnNfnNfNfNfnNfnNfnNfnNfnNfnNfsnsnN- moiety-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 5S: 5’-nsnsnnnNfnNfNfNfnnnnnnnnsnsnN-moiety-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the moiety may include any moiety such as a lipid moiety.
  • the sense strand comprises modification pattern 6S: 5’-nnnnNfNfimNfNfnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 7S: 5’- nnnnnnNfNfNfimnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 8S: 5’- nnnnnNfNfNfimnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 9S: 5’- nnnnnnNfiiNfNfimnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern IOS: 5’- nnnnnnnNfNfiiNfnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 1 IS: 5’- nnnnnNfnnNfimnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 12S: 5’- nnnnNfNfiiNfNfimnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 13S: 5’- nnnnNfnnnNfNfnnnnnnnnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 14S: 5’- nnnnNfnnNfNfnnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 15S: 5’- nnnnnNfNfNfNfNfnnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 16S: 5’- nnnnNfNfimNfiiNfhnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 17S: 5’- nnnnnNfNfiiNfiiNfhnnnnnnnnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 18S: 5’- nnnnNfiiNfiiNfiiNfhnnnnnnnnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 19S: 5’- nnnnNfiiNfirNfNfimnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 20S: 5’- nnnnnnnNfhNfimnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 21S: 5’- nnnnNfnnNfNfnNfimnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 22S: 5’- nnnnNfnnnNfiiNfnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 23 S: 5’- nnnnnNfnNfNfnnnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 24S: 5’- nnnnnnNfnNfhNfnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 25 S: 5’- nnnnnNfnNfNfnNfhnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 26S: 5’- nnnnnnnNfhnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 27S: 5’- nnnnNfnNfhNfnnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 28S: 5’- nnnnnNfnnNfNfnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 29S: 5’- nnnnnNfnnNfiiNfnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 30S: 5’- nnnnNfNfhnNfnnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 3 IS: 5’- nnnnnNfNfhNfnnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 32S: 5’- nnnnnNfNfhNfNfimnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 33 S: 5’- nnnnnnnNfNfNfimnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 34S: 5’- nnnnnnNfNfNfNfnnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 35 S: 5’- nnnnnNfiiNfNfNfnnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 36S: 5’- nnnnnNfNfNfhNfnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 37S: 5’- nnnnNfnnNfNfNfNfnnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 38S: 5’- nnnnNfiiNfNfNfimnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 39S: 5’- nnnnNfNfhNfNfiiNfnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 40S: 5’- nnnnNfNfNfNfNfnnnnnnnnsnsn-3 ’ , wherein “Nf’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O- methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 4 IS: 5’- nnnnNfiiNfNfdNnnnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 42S: 5’-NfsnsNfnNfnNfnNfnNfnNfnNfnNfiiNfiiNfsn-3’, wherein “Nf’ is a 2’-fluoro- modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 43S: 5’-snnnnNfNfimNfNfimnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 44S: 5’-snnnnnnNfhNfNfnnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 45 S: 5’- snnnnnNfhNfNfnNfnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 46S: 5’- snnnnNfnNfNfdNnnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 47S: 5’- snnnnnNfnnNfnnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 48S: 5’- snnnnNfNfnNfimnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 49S: 5’- snnnnNfnnnNfNfimnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 50S: 5’- snnnnNfNfhNfNfiiNfnnnnnnnnsnsn-3’, wherein “Nf ’ is a 2 ’-fluoro -modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 5 IS: 5’- snnnnNfnNfnNfNfhnnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 52S: 5’- snnnnnNfNfnNfiiNfhnnnnnnnnsnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 53 S: 5’- snnnnNfnNfiiNfiiNfhnnnnnnnnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 54S: 5’- snnnnnnNfNfNfimnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 55 S: 5’ - snnnnnnnnNfiiNfhnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 56S: 5’- snnnnnnnNfNfNfimnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 57S: 5’- snnnnNfnnNfNfnnnnnnnnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 58S: 5’ - snnnnnNfhNfNfimnnnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 59S: 5’- snnnnNfiiNfiiNfimnnnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 60S: 5’- snnnnnNfimNfNfnnnnnnnnsn- 3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’- O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 6 IS: 5’- snnnNmnNfNfNfNfnnnNmnnnnnnnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “Nm” is a 2’-O-methoxyethyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 62S: 5’- snnnNmnNfNfNfnnNmnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “Nm” is a 2’-O-methoxyethyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 63 S: 5’- snnnNmnNfNfNfNfnnnnNmnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “Nm” is a 2’-O-methoxyethyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 64S: 5’- snnnnNmNfNfNfnnnNmnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “Nm” is a 2’-O-methoxyethyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 65S: 5’- snnnnNmNfNfNfnnNmnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “Nm” is a 2’-O-methoxyethyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 66S: 5’- snnnnNmNfNfNfNfnnnnNmnnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “Nm” is a 2’-O-methoxyethyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 67S: 5’- snnnNmnNfNfNfnNmnnnnnnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “Nm” is a 2’-O-methoxyethyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the sense strand comprises modification pattern 68S: 5’- snnnNmnNfNfNfNfnnnnnnNmnnnnnsn-3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “Nm” is a 2’-O-methoxyethyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides. In some embodiments, the “Nm” is a 2’-O-methoxyethyl modified thymine.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2 wherein the oligonucleotide comprises an siRNA comprising a sense strand and an antisense strand, wherein the antisense strand comprises modification pattern IAS: 5’-nsNfsnNfiiNfiiNfiiNfnnnNfnNfiiNfnsn-3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, and “s” is a phosphorothioate or phosphate linkage.
  • the antisense strand comprises modification pattern 2AS: 5’-nsNfsnnnNfiiNfNfimnnNfhNfnnnsnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’- O-methyl modified nucleoside, and “s” is a phosphorothioate or phosphate linkage.
  • the antisense strand comprises modification pattern 3 AS: 5’-nsNfsnnnNfnnnnnnnnNfiiNfnnnsn-3’, wherein “Nf ’ is a 2’-fhioro-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, and “s” is a phosphorothioate or phosphate linkage.
  • the antisense strand comprises modification pattern 4AS: 5’-nsNfsnNfiiNfnnnnnnnNfhNfimnsnsn-3’, wherein “Nf ’ is a 2’-fluoro- modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, and “s” is a phosphorothioate or phosphate linkage.
  • the antisense strand comprises modification pattern 5 AS: 5’-nsNfsnNfiiNfiiNfiiNfnNfnNfhNfnNfnsn-3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, and “s” is a phosphorothioate or phosphate linkage.
  • the antisense strand comprises modification pattern 6AS:
  • Nf is a 2’-fhioro-modified nucleoside
  • n is a 2’- O-methyl modified nucleoside
  • s is a phosphorothioate or phosphate linkage
  • the antisense strand comprises modification pattern 7AS: 5 ’ -nsNfsnNfhNfiiNfnNfiiNfnNfiiNfiiNfiisnsn - 3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’- O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 8AS: 5’- nsnsnNfiiNfiiNfiiNfiiNfiiNfiiNfhNfhsnsn -3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 9AS: 5’-nsNfsnnnNfhNfimnnnNfiiNfnnnsn -3’, wherein “Nf’ is a 2’- fhioro -modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 10AS: 5’- nsNfsnnnNfiiNfnNfnNfiiNfiiNfnsnsn -3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 1 IAS: 5’-nsNfsnNfimNfhNfnnnnNfhNfnNfnsnsn -3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 12AS: 5’- nsNfsnNfnnNfnnNfnNfiiNfiiNfnsnsn -3’, wherein “Nf’ is a 2’-fhioro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 13AS: 5’-nsNfsnNfimNfimNfnNfiiNfimnnnsnsn -3’, wherein “Nf’ is a 2’- fluoro -modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 14AS: 5’- nsNfsnnnNfhNfnNfnNfiiNfiiNfimnsnsn -3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 15AS: 5’-nsNfsnnnNfiiNfiiNfiiNfnnnsn -3’, wherein “Nf’ is a 2’- fluoro -modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 16AS: 5’- nsNfsnNfnnNfnnnnNfnNfiiNfnNfiisnsn -3’, wherein “Nf ’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the antisense strand comprises modification pattern 17AS: 5’-nsNfsnnNfiiNfiiNfnnnnNfiiNfnsnsn -3’, wherein “Nf’ is a 2’-fluoro-modified nucleoside, “dN” is a 2’ deoxy-modified nucleoside, “n” is a 2’-O-methyl modified nucleoside, “s” is a phosphorothioate or phosphate linkage, and N comprises one or more nucleosides.
  • the sense strand comprises pattern ISand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, I 3AS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 2Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. 14AS, 15 AS, 16AS, or 17AS.
  • the sense strand comprises pattern 3 Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 4Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. WAS, 15AS, 16AS, or WAS.
  • the sense strand comprises pattern 5 Sand the antisense pattern comprises pattern IAS, 2AS, 3 AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. 14AS, 15AS, 16AS, or WAS.
  • the sense strand comprises pattern 6Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or WAS.
  • the sense strand comprises pattern 7Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or WAS.
  • the sense strand comprises pattern 8Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or WAS.
  • the sense strand comprises pattern 9Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or WAS.
  • the sense strand comprises pattern lOSand the antisense pattern comprises pattern IAS, 2AS, 3 AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern 11 Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern 12Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern BSand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern 14Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern BSand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern 16Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern WSand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern BSand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern WSand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern 20Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern 21 Sand the antisense pattern comprises pattern IAS, 2AS, 3 AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or WAS.
  • the sense strand comprises pattern 22Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, 13AS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 23 Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, I 3AS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 24Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, I 3AS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 25Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, I 3AS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 26Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 27Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 28Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 29Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 30Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 31 Sand the antisense pattern comprises pattern IAS, 2AS, 3 AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 32Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 33Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 34Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or 17AS.
  • the sense strand comprises pattern 35Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or 17AS.
  • the sense strand comprises pattern 36Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or 17AS.
  • the sense strand comprises pattern 37Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or 17AS.
  • the sense strand comprises pattern 38Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or 17AS.
  • the sense strand comprises pattern 39Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or 17AS.
  • the sense strand comprises pattern 40Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, 13AS, WAS, 15AS, 16AS, or 17AS.
  • the sense strand comprises patern 41Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. HAS. 15AS, 16AS, or I 7AS.
  • the sense strand comprises patern 42Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. 14AS, 15AS, 16AS, or I 7AS.
  • the sense strand comprises patern 43
  • Sand the antisense patern comprises patern IAS, 2AS, 3 AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, 15AS, 16AS, or I 7AS.
  • the sense strand comprises patern 44Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, 15AS, 16AS, or I 7AS.
  • the sense strand comprises patern 45
  • Sand the antisense patern comprises patern IAS, 2AS, 3 AS, 4AS, 5 AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, 15AS, 16AS, or I 7AS.
  • the sense strand comprises patern 46Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 47Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 48Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 49Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 50Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 5 ISand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 52Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 53Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 54Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 55Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 56Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 57Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS, 14AS, HAS, 16AS, or WAS.
  • the sense strand comprises patern 58Sand the antisense patern comprises patern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, 13AS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 59Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, I 3AS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 60Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, I 3AS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 61 Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, I 3AS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 62Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 63Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, HAS. 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 64Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 65Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 1 IAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 66Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 67Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, 14AS, 15AS, 16AS, or 17AS.
  • the sense strand comprises pattern 68Sand the antisense pattern comprises pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, BAS, 14AS, BAS, 16AS, or 17AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS,
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 2AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 3AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 11 S, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53 S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 4AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 5AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 6AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 7AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 8AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 9AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 10AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 1 IAS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 12AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 5 IS, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 13AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern MAS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 15AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 16AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S, and the antisense strand comprises pattern 17AS.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 5 IS, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S.
  • the sense strand comprises pattern IS, 2S, 3S, 4S, or 5 S.
  • the antisense strand comprises modification pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, 1 IAS, 12AS, BAS, WAS, 15AS, 16AS, or WAS.
  • the sense strand comprises modification pattern IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, 10AS, HAS, 12AS, BAS, WAS, 15AS, 16AS, or WAS.
  • the antisense strand comprises modification pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, IOS, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 51S, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S60S, 61S, 62S, 63S, 64S, 65S, 66S, 67S, or 68S
  • purines of the sense strand comprise 2’ -fluoro modified purines. In some embodiments, purines of the sense strand comprise 2’-O-methyl modified purines. In some embodiments, purines of the sense strand comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified purines. In some embodiments, all purines of the sense strand comprise 2’-fluoro modified purines. In some embodiments, all purines of the sense strand comprise 2’-O-methyl modified purines. In some embodiments, all purines of the sense strand comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified purines.
  • pyrimidines of the sense strand comprise 2’-fluoro modified pyrimidines. In some embodiments, pyrimidines of the sense strand comprise 2’-O-methyl modified pyrimidines. In some embodiments, pyrimidines of the sense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the sense strand comprise 2 ’-fluoro modified pyrimidines. In some embodiments, all pyrimidines of the sense strand comprise 2’-O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the sense strand comprise a mixture of 2’- fluoro and 2’-O-methyl modified pyrimidines.
  • purines of the sense strand comprise 2’-fluoro modified purines, and pyrimidines of the sense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, purines of the sense strand comprise 2’-O-methyl modified purines, and pyrimidines of the sense strand comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, purines of the sense strand comprise 2’ -fluoro modified purines, and pyrimidines of the sense strand comprise 2’-O-methyl modified pyrimidines.
  • purines of the sense strand comprise 2’-O-methyl modified purines
  • pyrimidines of the sense strand comprise 2’-fluoro modified pyrimidines.
  • pyrimidines of the sense strand comprise 2 ’-fluoro modified pyrimidines
  • purines of the sense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines.
  • pyrimidines of the sense strand comprise 2’-O-methyl modified pyrimidines
  • purines of the sense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines.
  • pyrimidines of the sense strand comprise 2 ’-fluoro modified pyrimidines, and purines of the sense strand comprise 2’-O-methyl modified purines. In some embodiments, pyrimidines of the sense strand comprise 2’-O-methyl modified pyrimidines, and purines of the sense strand comprise 2’ -fluoro modified purines.
  • all purines of the sense strand comprise 2’-fluoro modified purines, and all pyrimidines of the sense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the sense strand comprise 2’-O-methyl modified purines, and all pyrimidines of the sense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the sense strand comprise 2 ’-fluoro modified purines, and all pyrimidines of the sense strand comprise 2’-O-methyl modified pyrimidines.
  • all purines of the sense strand comprise 2’-O-methyl modified purines, and all pyrimidines of the sense strand comprise 2’-fluoro modified pyrimidines. In some embodiments, all pyrimidines of the sense strand comprise 2’-fluoro modified pyrimidines, and all purines of the sense strand comprise a mixture of 2’- fluoro and 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the sense strand comprise 2’-O-methyl modified pyrimidines, and all purines of the sense strand comprise a mixture of 2’- fluoro and 2’-O-methyl modified purines.
  • all pyrimidines of the sense strand comprise 2’-fluoro modified pyrimidines, and all purines of the sense strand comprise 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the sense strand comprise 2’-O-methyl modified pyrimidines, and all purines of the sense strand comprise 2 ’-fluoro modified purines.
  • purines of the antisense strand comprise 2’-fluoro modified purines. In some embodiments, purines of the antisense strand comprise 2’-O-methyl modified purines. In some embodiments, purines of the antisense strand comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified purines. In some embodiments, all purines of the antisense strand comprise 2 ’-fluoro modified purines. In some embodiments, all purines of the antisense strand comprise 2’-O-methyl modified purines. In some embodiments, all purines of the antisense strand comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified purines.
  • pyrimidines of the antisense strand comprise 2 ’-fluoro modified pyrimidines. In some embodiments, pyrimidines of the antisense strand comprise 2’-O-methyl modified pyrimidines. In some embodiments, pyrimidines of the antisense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the antisense strand comprise 2’-fluoro modified pyrimidines. In some embodiments, all pyrimidines of the antisense strand comprise 2’-O-methyl modified pyrimidines. In some embodiments, all pyrimidines of the antisense strand comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified pyrimidines.
  • purines of the antisense strand comprise 2’-fluoro modified purines, and pyrimidines of the antisense strand comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, purines of the antisense strand comprise 2’-O-methyl modified purines, and pyrimidines of the antisense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, purines of the antisense strand comprise 2’-fluoro modified purines, and pyrimidines of the antisense strand comprise 2’-O-methyl modified pyrimidines.
  • purines of the antisense strand comprise 2’-O-methyl modified purines
  • pyrimidines of the antisense strand comprise 2’-fluoro modified pyrimidines.
  • pyrimidines of the antisense strand comprise 2 ’-fluoro modified pyrimidines
  • purines of the antisense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines.
  • pyrimidines of the antisense strand comprise 2’-O-methyl modified pyrimidines
  • purines of the antisense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines.
  • pyrimidines of the antisense strand comprise 2’-fluoro modified pyrimidines, and purines of the antisense strand comprise 2’- O-methyl modified purines. In some embodiments, pyrimidines of the antisense strand comprise 2’-O- methyl modified pyrimidines, and purines of the antisense strand comprise 2’-fluoro modified purines. [00178] In some embodiments, all purines of the antisense strand comprise 2’-fluoro modified purines, and all pyrimidines of the antisense strand comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified pyrimidines.
  • all purines of the antisense strand comprise 2’-O-methyl modified purines, and all pyrimidines of the antisense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the antisense strand comprise 2 ’-fluoro modified purines, and all pyrimidines of the antisense strand comprise 2’-O-methyl modified pyrimidines. In some embodiments, all purines of the antisense strand comprise 2’-O-methyl modified purines, and all pyrimidines of the antisense strand comprise 2 ’-fluoro modified pyrimidines.
  • all pyrimidines of the antisense strand comprise 2 ’-fluoro modified pyrimidines, and all purines of the antisense strand comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the antisense strand comprise 2’-O-methyl modified pyrimidines, and all purines of the antisense strand comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the antisense strand comprise 2’-fluoro modified pyrimidines, and all purines of the antisense strand comprise 2’-O-methyl modified purines. In some embodiments, all pyrimidines of the antisense strand comprise 2’-O-methyl modified pyrimidines, and all purines of the antisense strand comprise 2 ’-fluoro modified purines.
  • modified oligonucleotides may be an siRNA that includes modifications to the ribose rings, and phosphate linkages. The modifications may be in particular patterns that maximize cell delivery, stability, and efficiency.
  • the siRNA may also include a vinyl phosphonate and a hydrophobic group. These modifications may aid in delivery to a cell or tissue within a subject.
  • the modified oligonucleotide may be used in a method such as a treatment method or a method of reducing gene expression.
  • the siRNA comprises a sense strand, an antisense strand, and a lipid moiety connected to an end of the sense or antisense strand; wherein the lipid moiety comprises a phenyl or cyclohexanyl linker, wherein the linker is connected to a lipid and to the end of the sense or antisense strand.
  • any one of the following is true with regard to the sense strand: (a) all purines comprise fluoro modified purines and all pyrimidines comprise (i) a mixture of 2’-O-methyl and 2’-O-methoxyethyl modified pyrimidines; or (ii) a mixture of 2’-fluoro, 2’-O-methyl, and 2’-O- methoxyethyl modified pyrimidines; (b) all purines comprise 2'-O-methyl modified purines and all pyrimidines comprise (i) all pyrimidines of the sense strand comprise a mixture of 2 ’-fluoro and 2’-O- methoxyethyl modified pyrimidines; or (ii) a mixture of 2’-fluoro, 2’-O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (c) all purines comprise 2'-O-methoxyethyl modified purines and all
  • any one of the following is true with regard to the antisense strand: all purines comprise 2’-fluoro modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; all purines comprise 2’-O-methyl modified purines, and all pyrimidines comprise 2’ -fluoro modified pyrimidines; all pyrimidines comprise 2’ -fluoro modified pyrimidines, and all purines comprise a mixture of 2’-fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O-methyl modified pyrimidines, and all purines comprise a mixture of 2 ’-fluoro and 2’-O-methyl modified purines; all pyrimidines comprise 2’-O-methyl modified
  • the siRNA comprises comprising a sense strand and an antisense strand; wherein the antisense strand comprises a 5’ end comprising a vinyl phosphonate and 2 phosphorothioate linkages, and a 3’ end comprising 2 phosphorothioate linkages; wherein the sense strand comprises (a) all purines comprise fluoro modified purines and all pyrimidines comprise (i) a mixture of 2’-O-methyl and 2’-O-methoxyethyl modified pyrimidines; or (ii) a mixture of 2’-fluoro, 2’-O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (b) all purines comprise 2'-O- methyl modified purines and all pyrimidines comprise (i) all pyrimidines of the sense strand comprise a mixture of 2’-fluoro and 2’-O-methoxyethyl modified pyrimidines; or (i
  • all purines comprise a mixture of 2’-fluoro and 2'-O-methoxyethyl modified purines and all pyrimidines of the sense strand comprise (i) 2’-O-methyl modified pyrimidines; (ii) a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; (iii) a mixture of 2’-O-methyl and 2’-O-methoxyethyl modified pyrimidines; or (iv) a mixture of 2’-fluoro, 2’-O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (f) all purines comprise a mixture of 2'-O-methyl and 2'-O-methoxyethyl modified purines and all pyrimidines comprise (i) 2’-fluoro modified pyrimidines; (ii) a mixture of 2’-fluoro and 2’-O-methyl modified pyrimidines; (ii)
  • any one of the following is true with regard to the sense strand: (a) all purines comprise fluoro modified purines and all pyrimidines comprise (i) a mixture of 2’-O-methyl and 2’-O-methoxyethyl modified pyrimidines; or (ii) a mixture of 2’-fluoro, 2’-O-methyl, and 2’-O- methoxyethyl modified pyrimidines; (b) all purines comprise 2'-O-methyl modified purines and all pyrimidines comprise (i) all pyrimidines of the sense strand comprise a mixture of 2 ’-fluoro and 2’-O- methoxyethyl modified pyrimidines; or (ii) a mixture of 2’-fluoro, 2’-O-methyl, and 2’-O-methoxyethyl modified pyrimidines; (c) all purines comprise 2'-O-methoxyethyl modified pur
  • a deoxy nucleoside may be included in the sense strand.
  • the sense strand includes the deoxy nucleoside.
  • the deoxy nucleoside may be at nucleoside position 9 of the sense strand.
  • the sense strand does not include a deoxy nucleoside.
  • the deoxy nucleoside of the sense strand may be otherwise unmodified.
  • the oligonucleotide comprises a duplex consisting of 21 nucleotide single strands with base pairing between 19 of the base pairs.
  • the duplex comprises single-stranded 2 nucleotide overhangs are at the 3’ ends of each strand.
  • One strand (antisense strand) is complementary to a DKK2 mRNA.
  • Each end of the antisense strand has one to two phosphorothioate bonds.
  • the 5’ end has an optional phosphate mimic such as a vinyl phosphonate.
  • the oligonucleotide is used to knock down a DKK2 mRNA or a target protein.
  • the sense strand has the same sequence as the DKK2 mRNA. In some embodiments, there are 1-2 phosphorothioates at the 3’ end. In some embodiments, there are 1 or no phosphorothioates at the 5’ end. In some embodiments, there is a hydrophobic conjugate of 12 to 25 carbons attached at the 5’ end via a phosphodiester bond.
  • the sense strand of any of the siRNAs comprises siRNA with a particular modification pattern.
  • position 9 counting from the 5’ end of the sense strand may have a 2’F modification.
  • position 9 of the sense strand is a pyrimidine, then all purines in the sense strand have a 2’0Me modification.
  • position 9 is the only pyrimidine between positions 5 and 11 of the sense stand, then position 9 is the only position with a 2’F modification in the sense strand.
  • both of these pyrimidines are the only two positions with a 2’F modification in the sense strand.
  • position 9 and only two other bases between positions 5 and 11 of the sense strand are pyrimidines, and those two other pyrimidines are in adjacent positions so that there would be not three 2’F modifications in a row, then any combination of 2’F modifications can be made that give three 2’F modifications in total.
  • the sense strand of any of the siRNAs comprises a modification pattern which conforms to any or all of these sense strand rules.
  • position 9 of the sense strand when position 9 of the sense strand is a purine, then all purines in the sense strand have a 2’0Me modification. In some embodiments, when position 9 is the only purine between positions 5 and 11 of the sense stand, then position 9 is the only position with a 2’F modification in the sense strand. In some embodiments, when position 9 and only one other base between positions 5 and 11 of the sense strand are purines, then both of these purines are the only two positions with a 2’F modification in the sense strand.
  • any combination of 2’F modifications can be made that give three 2’F modifications in total.
  • all combinations of purines having the 2’F modification are allowed that have three to five 2’F modifications in total, provided that the sense strand does not have three 2’F modifications in a row.
  • the sense strand of any of the siRNAs comprises a modification pattern which conforms to any or all of these sense strand rules.
  • position 9 of the sense strand can be a 2’deoxy.
  • 2’F and 2’0Me modifications may occur at the other positions of the sense strand.
  • the sense strand of any of the siRNAs comprises a modification pattern which conforms to these sense strand rules.
  • the sense strand comprises or consists of RNA or modified RNA nucleotides.
  • the sense strand comprises a deoxy nucleoside.
  • the deoxy nucleoside may include a DNA nucleoside.
  • the deoxy nucleoside comprises or consists of a 2’ deoxy nucleoside.
  • the deoxy nucleoside may be at a position within the sense strand (5’ to 3’, where the 5’ position is 1).
  • the position within the sense strand may be or include position 2, 4, 6, 8, 9, 10, 12, 14, 16, or 18, or a combination of said positions.
  • the position within the sense strand may be or include position 2, 4, 6, 8, 10, 12, 14, 16, or 18, or a combination of said positions.
  • the position within the sense strand may be or include position 2, 6, 9, 10, 14, or 18, or a combination of said positions.
  • the position within the sense strand may be or include position 2, 6, 10, 14, or 18, or a combination of said positions.
  • the position within the sense strand may be or include position 4, 8, 9, 12, or 16, or a combination of said positions.
  • the position within the sense strand may be or include position 4, 8, 12, or 16, or a combination of said positions.
  • the position within the sense strand may include position 9.
  • the position within the sense strand may be position 9.
  • the sense strand may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 deoxy nucleosides. In some embodiments, the sense strand includes 1 deoxy nucleoside.
  • the sense strand may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 deoxy nucleosides, or a range of deoxy nucleosides defined by any two of the aforementioned numbers of deoxy nucleosides.
  • the sense strand may include deoxy nucleosides at all even positions.
  • the sense strand may include deoxy nucleosides at some even positions.
  • the sense strand may include deoxy nucleosides at every other even position.
  • the sense strand may include 1 deoxy nucleoside.
  • the sense strand may include at least 1 deoxy nucleoside.
  • the sense strand may include at least 2 deoxy nucleosides.
  • the sense strand may include at least 3 deoxy nucleosides.
  • the sense strand may include at least 4 deoxy nucleosides.
  • the sense strand may include at least 5 deoxy nucleosides.
  • the sense strand may include at least 6 deoxy nucleosides.
  • the sense strand may include at least 7 deoxy nucleosides.
  • the sense strand may include at least 8 deoxy nucleosides.
  • the sense strand may include at least 9 deoxy nucleosides.
  • the sense strand may include at least 10 deoxy nucleosides.
  • the sense strand may include no greater than 2 deoxy nucleosides.
  • the sense strand may include no greater than 3 deoxy nucleosides.
  • the sense strand may include no greater than 4 deoxy nucleosides.
  • the sense strand may include no greater than 5 deoxy nucleosides.
  • the sense strand may include no greater than 6 deoxy nucleosides.
  • the sense strand may include no greater than 7 deoxy nucleosides.
  • the sense strand may include no greater than 8 deoxy nucleosides.
  • the sense strand may include no greater than 9 deoxy nucleosides.
  • the sense strand may include no greater than 10 deoxy nucleosides.
  • the antisense strand comprises or consists of RNA or modified RNA nucleotides.
  • the antisense strand comprises a deoxy nucleoside.
  • the deoxy nucleoside may include a DNA nucleoside.
  • the deoxy nucleoside comprises or consists of a 2’ deoxy nucleoside.
  • the antisense strand may include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 deoxy nucleosides, or a range of deoxy nucleosides defined by any two of the aforementioned numbers of deoxy nucleosides.
  • nucleosides at positions 1-8 include a mixture of 2’-fluoro and 2’-O- methyl modified nucleosides.
  • purines at positions 1-8 include a mixture of 2 ’-fluoro and 2’-O-methyl modified nucleosides.
  • pyrimidines at positions 1-8 include a mixture of 2 ’-fluoro and 2’-O-methyl modified nucleosides.
  • nucleosides at positions 1-8 all include 2’-O-methyl modified nucleosides.
  • purines at positions 1-8 all include 2’-O-methyl modified nucleosides.
  • pyrimidines at positions 1-8 all include 2’-O-methyl modified nucleosides.
  • purines at positions 1-8 include a mixture of 2 ’-fluoro and 2’-O-methyl modified nucleosides, and pyrimidines at positions 1-8 all include 2’-O-methyl modified nucleosides.
  • pyrimidines at positions 1-8 include a mixture of 2’-fluoro and 2’-O-methyl modified nucleosides, and purines at positions 1-8 all include 2’-O-methyl modified nucleosides.
  • compositions comprising an oligonucleotide that targets DKK2 and when administered to a cell decreases expression of DKK2, wherein the oligonucleotide comprises a small interfering RNA (siRNA) comprising a sense strand and an antisense strand, wherein the sense strand comprises a sense strand sequence described herein in which at least one intemucleoside linkage is modified and at least one nucleoside is modified, or an sense strand sequence comprising 1 or 2 nucleoside substitutions, additions, or deletions of the oligonucleotide sequence in which at least one intemucleoside linkage is modified and at least one nucleoside is modified, and wherein the antisense strand comprises an antisense strand sequence described herein in which at least one intemucleoside linkage is modified and at least one nucleoside is modified, or an oligonucleotide sequence comprising 1 or 2 nucleo
  • siRNA small interfering RNA
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence in Table 4.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence in Table 4.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 4, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 4, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence in Table 4.
  • the sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand may comprise an overhang.
  • the sense strand or antisense strand may comprise any modifications described herein (e.g. a different set of modifications or modification pattern than in Table 4).
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence in Table 5.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence in Table 5.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 5, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 5, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence in Table 5.
  • the sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand may comprise an overhang.
  • the sense strand or antisense strand may comprise any modifications described herein (e.g. a different set of modifications or modification pattern than in Table 5).
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence in Table 6.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence in Table 6.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 6, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 6, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence in Table 6.
  • the sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
  • the sense strand or antisense strand may comprise an overhang.
  • the sense strand or antisense strand may comprise any modifications described herein (e.g. a different set of modifications or modification pattern than in Table 6).
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence in Table 13A.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence in Table 13A.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 13A, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 13A, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence in Table 13A.
  • the sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
  • the sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand may comprise an overhang.
  • the sense strand or antisense strand may comprise any modifications described herein (e.g. a different set of modifications or modification pattern than in Table 13A).
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • the siRNA comprises a sense strand or antisense strand having a sequence in accordance with the sense strand or antisense strand sequence in Table 17.
  • the sense strand or antisense strand comprises a sequence at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, or at least 95% identical, to a sense strand or antisense strand sequence in Table 17.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 17, or a sequence thereof having 3 or 4 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense strand comprises a sequence of a sense or antisense strand in Table 17, or a sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions.
  • the sense strand or antisense sequence comprises or consists of a sequence 100% identical to a sense strand or antisense strand sequence in Table 17.
  • the sense strand or antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the first 19 nucleotides of any of the aforementioned sequences.
  • the sense strand or antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand or antisense strand may comprise an overhang.
  • the sense strand or antisense strand may comprise any modifications described herein (e.g. a different set of modifications or modification pattern than in Table 17).
  • the sense strand or antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • the siRNA comprises a sense strand having a sequence in accordance with any of SEQ ID Nos: 7666-7717.
  • the sense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID Nos: 7666-7717, at least 80% identical to any one of SEQ ID Nos: 7666-7717, at least 85% identical to of any one of SEQ ID Nos: 7666-7717, at least 90% identical to any one of SEQ ID Nos: 7666-7717, or at least 95% identical to any one of SEQ ID Nos: 7666-7717.
  • the sense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 7666-7717, or a sense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 1-3636, or a sense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the sense strand sequence comprises or consists of a sequence 100% identical to SEQ ID Nos: 7666-7717.
  • the sense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the sense strand may comprise a modification pattern described herein.
  • the sense strand may comprise an overhang.
  • the sense strand may comprise a lipid moiety.
  • the sense strand may comprise a GalNAc moiety.
  • the siRNA comprises an antisense strand having a sequence in accordance with any of SEQ ID Nos: 7718-7769.
  • the antisense strand sequence comprises or consists of sequence at least 75% identical to any one of SEQ ID Nos: 7718-7769, at least 80% identical to any one of SEQ ID Nos: 7718-7769, at least 85% identical to of any one of SEQ ID Nos: 7718-7769, at least 90% identical to any one of SEQ ID Nos: 7718-7769, or at least 95% identical to any one of SEQ ID Nos: 7718-7769.
  • the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 7718-7769, or an antisense strand sequence thereof having 1, 2, 3, or 4 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of the sequence of any one of SEQ ID Nos: 7718-7769, or an antisense strand sequence thereof having 1 or 2 nucleoside substitutions, additions, or deletions. In some embodiments, the antisense strand sequence comprises or consists of a sequence 100% identical to SEQ ID Nos: 7718-7769.
  • the antisense strand sequence may include the first 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the antisense strand sequence may include the last 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 nucleotides (in the 5’ to 3’ direction) of any of the aforementioned sequences.
  • the antisense strand may comprise an overhang.
  • the antisense strand may comprise a modification pattern described herein.
  • the antisense strand may comprise a lipid moiety or a GalNAc moiety.
  • the composition comprises an oligonucleotide that inhibits the expression of DKK2, wherein the oligonucleotide comprises an antisense oligonucleotide (ASO).
  • ASO comprises modification pattern ASO1: 5’-nsnsnsnsnsdNsdNsdNsdNsdNsdNsdNsdNsdNsdNsdNsnsnsnsn-3’, wherein “dN” is any deoxynucleotide, “n” is a 2’-O-methyl or 2 ’-O-methoxyethyl -modified nucleoside, and “s” is a phosphorothioate or phosphate linkage.
  • the ASO comprises modification pattern IS, 2S, 3S, 4S, 5S, 6S, 7S, 8S, 9S, 10S, 1 IS, 12S, 13S, 14S, 15S, 16S, 17S, 18S, 19S, 20S, 21S, 22S, 23S, 24S, 25S, 26S, 27S, 28S, 29S, 30S, 31S, 32S, 33S, 34S, 35S, 36S, 37S, 38S, 39S, 40S, 41S, 42S, 43S, 44S, 45S, 46S, 47S, 48S, 49S, 50S, 5 IS, 52S, 53S, 54S, 55S, 56S, 57S, 58S, 59S, 60S, IAS, 2AS, 3AS, 4AS, 5AS, 6AS, 7AS, 8AS, 9AS, WAS, HAS, 12AS, I 3AS. 14AS, 15AS, 16AS, or 17AS.
  • the composition is a pharmaceutical composition. In some embodiments, the composition is sterile. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier comprises water. In some embodiments, the pharmaceutically acceptable carrier comprises a buffer. In some embodiments, the pharmaceutically acceptable carrier comprises a saline solution. In some embodiments, the pharmaceutically acceptable carrier comprises a cream. In some embodiments, the pharmaceutically acceptable carrier comprises a gel. In some embodiments, the pharmaceutically acceptable carrier comprises water, a buffer, a saline solution, a cream, or a gel. In some embodiments, the pharmaceutically acceptable carrier comprises contains a permeation enhancer. In some embodiments, the formulation contains pharmaceutically acceptable counterions to the oligonucleotides. In some embodiments, the pharmaceutically acceptable counterions increase membrane affinity. In some embodiments, the composition comprises a liposome.
  • the pharmaceutically acceptable carrier comprises liposomes, lipids, nanoparticles, proteins, protein-antibody complexes, peptides, cellulose, nanogel, or a combination thereof.
  • the composition is formulated for topical administration.
  • kits may include an oligonucleotide such as an siRNA described herein.
  • the oligonucleotide may be conjugated to a lipid moiety or to a sugar moiety.
  • the kit may include a lipid moiety.
  • the kit may include a sugar moiety.
  • the oligonucleotide may comprise nucleoside modifications or modified intemucleoside linkages.
  • the oligonucleotide may include any modifications described herein, such as modifications from a base sequence.
  • the kit may include a delivery reagent such as a needle.
  • the kit may include instructions for use, such as methods for use in a method described herein.
  • composition described herein are methods of administering a composition described herein to a subject. Some embodiments relate to use a composition described herein, such as administering the composition to a subject.
  • Some embodiments relate to a method of treating a disorder in a subject in need thereof. Some embodiments relate to use of a composition described herein in the method of treatment. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration treats the disorder in the subject. In some embodiments, the composition treats the disorder in the subject.
  • the treatment comprises prevention, inhibition, or reversion of the disorder in the subject.
  • Some embodiments relate to use of a composition described herein in the method of preventing, inhibiting, or reversing the disorder.
  • Some embodiments relate to a method of preventing, inhibiting, or reversing a disorder a disorder in a subject in need thereof.
  • Some embodiments include administering a composition described herein to a subject with the disorder.
  • the administration prevents, inhibits, or reverses the disorder in the subject.
  • the composition prevents, inhibits, or reverses the disorder in the subject.
  • Some embodiments relate to a method of preventing a disorder a disorder in a subject in need thereof. Some embodiments relate to use of a composition described herein in the method of preventing the disorder. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration prevents the disorder in the subject. In some embodiments, the composition prevents the disorder in the subject.
  • Some embodiments relate to a method of inhibiting a disorder a disorder in a subject in need thereof. Some embodiments relate to use of a composition described herein in the method of inhibiting the disorder. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration inhibits the disorder in the subject. In some embodiments, the composition inhibits the disorder in the subject.
  • Some embodiments relate to a method of reversing a disorder a disorder in a subject in need thereof. Some embodiments relate to use of a composition described herein in the method of reversing the disorder. Some embodiments include administering a composition described herein to a subject with the disorder. In some embodiments, the administration reverses the disorder in the subject. In some embodiments, the composition reverses the disorder in the subject. [00207]
  • the administration may be topical.
  • a composition or formulation described herein may be administered to a scalp.
  • the topical administration may include rubbing, brushing, swabbing, dabbing, or wiping.
  • the administration may be on a skin area of the subject.
  • the skin area may include hair.
  • the skin area may include hair loss.
  • the skin area may be at risk of hair loss.
  • the skin area may include an area of the head.
  • the skin area may include a scalp.
  • the skin area may include a scalp region.
  • the skin area may include a temporal region
  • the disorder includes hair loss.
  • the disorder is hair loss.
  • hair loss include androgenetic alopecia (male pattern baldness), alopecia areata, and non-scarring hair loss.
  • the disorder includes hair discoloration or graying.
  • the hair loss comprises male pattern baldness.
  • the hair loss comprises alopecia areata.
  • the hair loss comprises scarring hair loss.
  • the hair loss comprises non-scarring hair loss.
  • Some embodiments of the methods described herein include treatment of a subject.
  • subjects include vertebrates, animals, mammals, dogs, cats, cattle, rodents, mice, rats, primates, monkeys, and humans.
  • the subject is a vertebrate.
  • the subject is an animal.
  • the subject is a mammal.
  • the subject is a dog.
  • the subject is a cat.
  • the subject is a cattle.
  • the subject is a mouse.
  • the subject is a rat.
  • the subject is a primate.
  • the subject is a monkey.
  • the subject is an animal, a mammal, a dog, a cat, cattle, a rodent, a mouse, a rat, a primate, or a monkey. In some embodiments, the subject is a human. In some embodiments, the subject is male. In some embodiments, the subject is female.
  • the subject has a body mass index (BMI) of 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, or more, or a range defined by any two of the aforementioned integers.
  • the subject is overweight.
  • the subject has a BMI of 25 or more.
  • the subject has a BMI of 25-29.
  • the subject is obese.
  • the subject has a BMI of 30 or more.
  • the subject has a BMI of 30-39.
  • the subject has a BMI of 40-50.
  • the subject has a BMI of 25-50.
  • the subject is > 90 years of age. In some embodiments, the subject is > 85 years of age. In some embodiments, the subject is > 80 years of age. In some embodiments, the subject is > 70 years of age. In some embodiments, the subject is > 60 years of age. In some embodiments, the subject is > 50 years of age. In some embodiments, the subject is > 40 years of age. In some embodiments, the subject is > 30 years of age. In some embodiments, the subject is > 20 years of age. In some embodiments, the subject is > 10 years of age. In some embodiments, the subject is > 1 years of age. In some embodiments, the subject is > 0 years of age.
  • the subject is ⁇ 100 years of age. In some embodiments, the subject is ⁇ 90 years of age. In some embodiments, the subject is ⁇ 85 years of age. In some embodiments, the subject is ⁇ 80 years of age. In some embodiments, the subject is ⁇ 70 years of age. In some embodiments, the subject is ⁇ 60 years of age. In some embodiments, the subject is ⁇ 50 years of age. In some embodiments, the subject is ⁇ 40 years of age. In some embodiments, the subject is ⁇ 30 years of age. In some embodiments, the subject is ⁇ 20 years of age. In some embodiments, the subject is ⁇ 10 years of age. In some embodiments, the subject is ⁇ 1 years of age.
  • the subject is between 0 and 100 years of age. In some embodiments, the subject is between 20 and 90 years of age. In some embodiments, the subject is between 30 and 80 years of age. In some embodiments, the subject is between 40 and 75 years of age. In some embodiments, the subject is between 50 and 70 years of age. In some embodiments, the subject is between 40 and 85 years of age. In some embodiments, the subject is aging. In some embodiments, the subject is an adult.
  • the subject has a family history of hair loss. In some embodiments, the subject has hormone levels related to hair loss. In some embodiments, the subject has a thyroid disorder. In some embodiments, the subject is malnourished. In some embodiments, the subject has been subjected to environmental factors affecting hair loss. In some embodiments, the subject has subjected to physical stress. In some embodiments, the subject has subjected to emotional stress.
  • a baseline measurement is obtained from the subject prior to treating the subject.
  • the baseline measurement is a baseline hair loss measurement.
  • the baseline measurement is a baseline measurement of a symptom of hair loss.
  • baseline measurements include a baseline hair loss assessment score, a baseline total hair count, a baseline vellus hair count, a baseline non-vellus hair count, a baseline hair thickness measurement, a baseline hair density measurement, or a baseline number of hair follicles.
  • the baseline measurement may include a baseline hair color measurement.
  • the baseline measurement may include a baseline gene or protein level, a baseline DKK2 mRNA level, or a baseline DKK2 protein level.
  • the baseline measurement is obtained non-invasively. In some embodiments, the baseline measurement is obtained directly from the subject. In some embodiments, the baseline measurement is obtained by observation, for example by observation of the subject or of the subject’s tissue. In some embodiments, the baseline measurement is obtained noninvasively using an imaging device. In some embodiments, the baseline measurement is obtained using a photograph. In some embodiments, the baseline measurement is obtained using a phototrichogram. In some embodiments, the baseline measurement is obtained using a macrophotography analysis. In some embodiments, the baseline measurement is obtained using a questionnaire. [00217] In some embodiments, the baseline measurement is obtained invasively. In some embodiments, the baseline measurement is obtained in a sample from the subject.
  • the baseline measurement is obtained in a biopsy such as a scalp biopsy. In some embodiments, the baseline measurement is obtained in one or more histological tissue sections. In some embodiments, the baseline measurement is obtained by performing an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay, on the sample obtained from the subject. In some embodiments, the baseline measurement is obtained by an immunoassay, a colorimetric assay, or a fluorescence assay. In some embodiments, the baseline measurement is obtained by PCR.
  • the baseline measurement is a baseline questionnaire result.
  • the baseline measurement is a baseline hair loss hair loss assessment score.
  • the baseline questionnaire result comprises a baseline score such as a baseline hair loss assessment score.
  • the baseline questionnaire result is obtained from a questionnaire.
  • the baseline questionnaire result is obtained from multiple questionnaires.
  • the questionnaire is a Men’s Hair Growth Questionnaire (MHGQ).
  • the questionnaire is a Kingsley Alopecia Profde (KAP) questionnaire.
  • the questionnaire may include questions about hair growth that the subject rates. Non-limiting examples of such ratings may include strongly agree, agree, neither agree nor disagree, disagree, and strongly disagree, where each rating is assigned a value.
  • the baseline score may include a sum of each value.
  • the baseline hair loss assessment score is not based on a questionnaire.
  • the baseline hair loss assessment score is assessed by a medical practitioner.
  • the baseline hair loss assessment score includes a semi-quantitative hair visual hair score on a numerical scale such as 1-10.
  • the baseline measurement is a baseline hair count.
  • the baseline hair count is a baseline total hair count.
  • the baseline total hair count may include a baseline vellus hair count and a baseline non-vellus hair count.
  • the baseline hair count is a baseline vellus hair count.
  • the baseline hair count is a baseline non-vellus hair count.
  • the baseline hair count is determined in an area of skin.
  • the baseline hair count is normalized based on the area of skin.
  • the baseline hair count is assessed using photography.
  • the baseline hair count is assessed by phototrichogram.
  • the baseline hair count is assessed by a macrophotography analysis.
  • the baseline measurement is a baseline hair thickness measurement.
  • the baseline hair thickness measurement is determined in an area of skin.
  • the baseline hair thickness measurement comprises a width of an individual hair.
  • the baseline hair thickness measurement comprises widths of multiple individual hairs.
  • the baseline hair thickness measurement comprises an average of the widths of the multiple individual hairs.
  • the baseline hair thickness measurement comprises a median of the widths of the multiple individual hairs.
  • the baseline hair thickness measurement may include a baseline vellus hair thickness measurement.
  • the baseline hair thickness measurement may include a baseline non-vellus hair thickness measurement.
  • the baseline hair thickness measurement is assessed using photography.
  • the baseline hair thickness measurement is assessed by phototrichogram.
  • the baseline hair thickness measurement is assessed by a macrophotography analysis.
  • the baseline measurement is a baseline hair density measurement.
  • the baseline hair density measurement is determined in an area of skin.
  • the baseline hair density measurement comprises a number of hair in the area of skin.
  • the baseline hair density measurement comprises the number of hair in the area of skin divided by the area of skin.
  • the baseline hair density measurement may include a baseline vellus hair density measurement.
  • the baseline hair density measurement may include a baseline non-vellus hair density measurement.
  • the baseline hair density measurement is assessed using photography.
  • the baseline hair density measurement is assessed by phototrichogram.
  • the baseline hair density measurement is assessed by a macrophotography analysis.
  • the baseline measurement is a baseline number of hair follicles. In some embodiments, the baseline number of hair follicles is a baseline total number of hair follicles. In some embodiments, the baseline number of hair follicles include a baseline number of terminal hair follicles. In some embodiments, the baseline number of hair follicles include a baseline number of anagen hair follicles. In some embodiments, the baseline number of hair follicles include a baseline number of telogen hair follicles. In some embodiments, the baseline number of hair follicles include a baseline number of catagen hair follicles.
  • the baseline number of hair follicles include a baseline number of miniaturized hair follicles. In some embodiments, the baseline number of hair follicles include a baseline number of vellus miniaturized hair follicles. In some embodiments, the baseline number of hair follicles include a baseline number of vellus-like miniaturized hair follicles. In some embodiments, the baseline number of hair follicles include a baseline number of indeterminate hair follicles. In some embodiments, the baseline number of hair follicles is determined in an area of skin. In some embodiments, the baseline number of hair follicles is normalized based on the area of skin. In some embodiments, the baseline number of hair follicles is assessed in a biopsy, such as a scalp biopsy. In some embodiments, the baseline number of hair follicles is assessed using photography.
  • the baseline measurement is a baseline hair color measurement.
  • the baseline hair color measurement is determined in an area of skin.
  • the baseline hair color measurement comprises a color of an individual hair.
  • the baseline hair color measurement comprises colors of multiple individual hairs.
  • the baseline hair color measurement comprises an average of the colors of the multiple individual hairs.
  • the baseline hair color measurement comprises a median of the colors of the multiple individual hairs.
  • the baseline hair color measurement may include a baseline measurement of how gray the hair is, or how gray the hairs are.
  • the baseline hair color measurement may include a baseline measurement of how much color is in the hair, or how much color is in the hairs.
  • the baseline hair color measurement may include a baseline measurement of how white the hair is, or how white the hairs are.
  • the baseline hair color measurement may include a hair pigmentation measurement.
  • the baseline hair color measurement may include a hair contrast measurement.
  • the baseline hair color measurement may include a baseline vellus hair color measurement.
  • the baseline hair color measurement may include a baseline non-vellus hair color measurement.
  • the baseline hair color measurement is assessed using photography.
  • the baseline hair color measurement is assessed by phototrichogram.
  • the baseline hair color measurement is assessed by a macrophotography analysis.
  • the baseline hair color measurement may be a qualitative measurement.
  • the baseline hair color measurement may be a quantitative measurement.
  • the baseline hair color measurement may be a number, such as an amount of hair color.
  • the baseline hair color measurement may be a rate, such as a rate of hair color loss.
  • the baseline measurement is a baseline protein level.
  • the baseline protein level is a baseline [3-catenin protein level.
  • the baseline protein level is a baseline a-SMA protein level.
  • the baseline protein level is a baseline collagen protein level.
  • the collagen of the baseline collagen protein level is collagen I.
  • the collagen of the baseline collagen protein level is collagen III.
  • the baseline protein level is assessed in a baseline sample such as a baseline skin sample.
  • the baseline protein level is indicated as a mass or percentage of protein per sample weight.
  • the baseline protein level is indicated as a mass or percentage of protein per sample volume.
  • the baseline protein level is indicated as a mass or percentage of protein per total protein within the sample.
  • the baseline protein measurement is a baseline circulating protein measurement.
  • the baseline protein measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.
  • the baseline measurement is a baseline mRNA level.
  • the baseline mRNA level is a baseline [3-catenin mRNA level.
  • the baseline mRNA level is a baseline a-SMA mRNA level.
  • the baseline mRNA level is a baseline collagen mRNA level.
  • the collagen of the baseline collagen mRNA level is collagen I.
  • the collagen of the baseline collagen mRNA level is collagen III.
  • the baseline mRNA level is assessed in a baseline sample such as a baseline skin sample.
  • the baseline mRNA level is indicated as a mass or percentage of mRNA per sample weight.
  • the baseline mRNA level is indicated as a mass or percentage of mRNA per sample volume. In some embodiments, the baseline mRNA level is indicated as a mass or percentage of mRNA per total mRNA within the sample. In some embodiments, the baseline mRNA level is indicated as a mass or percentage of mRNA per total nucleic acids within the sample. In some embodiments, the baseline mRNA level is indicated relative to another mRNA level, such as an mRNA level of a housekeeping gene, within the sample. In some embodiments, the baseline mRNA measurement is obtained by an assay such as a polymerase chain reaction (PCR) assay. In some embodiments, the PCR comprises quantitative PCR (qPCR). In some embodiments, the PCR comprises reverse transcription of the mRNA.
  • PCR quantitative PCR
  • the baseline measurement is a baseline DKK2 protein measurement.
  • the baseline DKK2 protein measurement comprises a baseline DKK2 protein level.
  • the baseline DKK2 protein level is assessed in a baseline sample such as a baseline skin sample or a baseline fluid sample (e.g. blood, serum, or plasma).
  • the baseline DKK2 protein level is indicated as a mass or percentage of DKK2 protein per sample weight.
  • the baseline DKK2 protein level is indicated as a mass or percentage of DKK2 protein per sample volume.
  • the baseline DKK2 protein level is indicated as a mass or percentage of DKK2 protein per total protein within the sample.
  • the baseline DKK2 protein measurement is a baseline circulating DKK2 protein measurement.
  • the baseline DKK2 protein measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.
  • the baseline measurement is a baseline DKK2 mRNA measurement.
  • the baseline DKK2 mRNA measurement comprises a baseline DKK2 mRNA level.
  • the baseline DKK2 mRNA level is assessed in a baseline sample such as a baseline skin sample.
  • the baseline DKK2 mRNA level is indicated as a mass or percentage of DKK2 mRNA per sample weight.
  • the baseline DKK2 mRNA level is indicated as a mass or percentage of DKK2 mRNA per sample volume.
  • the baseline DKK2 mRNA level is indicated as a mass or percentage of DKK2 mRNA per total mRNA within the sample.
  • the baseline DKK2 mRNA level is indicated as a mass or percentage of DKK2 mRNA per total nucleic acids within the sample. In some embodiments, the baseline DKK2 mRNA level is indicated relative to another mRNA level, such as an mRNA level of a housekeeping gene, within the sample. In some embodiments, the baseline DKK2 mRNA measurement is obtained by an assay such as a polymerase chain reaction (PCR) assay. In some embodiments, the PCR comprises quantitative PCR (qPCR). In some embodiments, the PCR comprises reverse transcription of the DKK2 mRNA.
  • PCR polymerase chain reaction
  • Some embodiments of the methods described herein include obtaining a sample from a subject.
  • the baseline measurement is obtained in a sample obtained from the subject.
  • the sample is obtained from the subject prior to administration or treatment of the subject with a composition described herein.
  • a baseline measurement is obtained in a sample obtained from the subject prior to administering the composition to the subject.
  • the sample is obtained from the subject in a fasted state.
  • the sample is obtained from the subject after an overnight fasting period.
  • the sample is obtained from the subject in a fed state.
  • the sample comprises a fluid. In some embodiments, the sample is a fluid sample. In some embodiments, the sample is a blood, plasma, or serum sample. In some embodiments, the sample comprises blood. In some embodiments, the sample is a blood sample. In some embodiments, the sample is a whole-blood sample. In some embodiments, the blood is fractionated or centrifuged. In some embodiments, the sample comprises plasma. In some embodiments, the sample is a plasma sample. In some embodiments, the sample comprises serum. In some embodiments, the sample is a serum sample. [00230] In some embodiments, the sample comprises a tissue. The tissue may be or include skin. The skin may comprise or consist of a skin layer.
  • the skin layer may be or include a dermal layer or an epidermal layer.
  • the skin may include epidermis.
  • the skin may include epidermis.
  • the skin may include scalp skin.
  • the skin may eyebrow skin.
  • the skin may pubic skin.
  • the skin may include skin from a leg of the subject.
  • the skin may include skin from an arm of the subject.
  • the skin may include one or more hairs.
  • the hairs may be any of scalp hairs, eyebrow hairs, pubic hairs, arm hairs, or leg hairs.
  • the sample is a tissue sample.
  • the sample comprises skin.
  • the sample is a skin sample.
  • the baseline DKK2 mRNA measurement, or the baseline DKK2 protein measurement may be obtained in a skin sample from the patient prior to administration of a compound or oligonucleotide disclosed herein.
  • the sample is a biopsy.
  • the biopsy is a skin biopsy.
  • the skin biopsy includes a scalp biopsy.
  • the composition or administration of the composition affects a measurement such as a hair loss measurement or a measurement of a symptom of hair loss.
  • the measurement is a hair loss assessment score, a total hair count, a vellus hair count, a non-vellus hair count, a hair thickness measurement, a hair density measurement, a number of hair follicles, a hair color measurement, a gene or protein level, a DKK2 protein measurement (for example, circulating or tissue DKK2 protein levels), or a DKK2 mRNA measurement, relative to the baseline measurement.
  • Some embodiments of the methods described herein include obtaining the measurement from a subject.
  • the measurement may be obtained from the subject after treating the subject.
  • the measurement is obtained in a second sample (such as a fluid or tissue sample described herein) obtained from the subject after the composition is administered to the subject.
  • the measurement indicates that the disorder has been treated.
  • the measurement is obtained non-invasively. In some embodiments, the measurement is obtained directly from the subject. In some embodiments, the measurement is obtained by observation, for example by observation of the subject or of the subject’s tissue. In some embodiments, the measurement is obtained noninvasively using an imaging device. In some embodiments, the measurement is obtained using a photograph. In some embodiments, the measurement is obtained using a phototrichogram. In some embodiments, the measurement is obtained using a macrophotography analysis. In some embodiments, the measurement is obtained using a questionnaire.
  • the measurement is obtained invasively. In some embodiments, the measurement is obtained in a second sample from the subject. In some embodiments, the measurement is obtained in a biopsy such as a scalp biopsy. In some embodiments, the measurement is obtained in one or more histological tissue sections. In some embodiments, the measurement is obtained by performing an assay on the second sample obtained from the subject. In some embodiments, the measurement is obtained by an assay, such as an assay described herein. In some embodiments, the assay is an immunoassay, a colorimetric assay, a fluorescence assay, or a PCR assay.
  • the measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.
  • the measurement is obtained by PCR.
  • the measurement is obtained by histology.
  • the measurement is obtained by observation.
  • additional measurements are made, such as in a 3rd sample, a 4th sample, or a fifth sample.
  • the measurement is obtained within 1 hour, within 2 hours, within 3 hours, within 4 hours, within 5 hours, within 6 hours, within 12 hours, within 18 hours, or within 24 hours after the administration of the composition. In some embodiments, the measurement is obtained within 1 day, within 2 days, within 3 days, within 4 days, within 5 days, within 6 days, or within 7 days after the administration of the composition. In some embodiments, the measurement is obtained within 1 week, within 2 weeks, within 3 weeks, within 1 month, within 2 months, within 3 months, within 6 months, within 1 year, within 2 years, within 3 years, within 4 years, or within 5 years after the administration of the composition.
  • the measurement is obtained after 1 hour, after 2 hours, after 3 hours, after 4 hours, after 5 hours, after 6 hours, after 12 hours, after 18 hours, or after 24 hours after the administration of the composition. In some embodiments, the measurement is obtained after 1 day, after 2 days, after 3 days, after 4 days, after 5 days, after 6 days, or after 7 days after the administration of the composition. In some embodiments, the measurement is obtained after 1 week, after 2 weeks, after 3 weeks, after 1 month, after 2 months, after 3 months, after 6 months, after 1 year, after 2 years, after 3 years, after 4 years, or after 5 years, following the administration of the composition.
  • the composition reduces the measurement relative to the baseline measurement.
  • the reduction is measured in a second tissue sample obtained from the subject after administering the composition to the subject.
  • the reduction is measured directly in the subject after administering the composition to the subject.
  • the measurement is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline measurement.
  • the measurement is decreased by about 10% or more, relative to the baseline measurement.
  • the measurement is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline measurement.
  • the measurement is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline measurement. In some embodiments, the measurement is decreased by no more than about 10%, relative to the baseline measurement. In some embodiments, the measurement is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline measurement. In some embodiments, the measurement is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.
  • the composition increases the measurement relative to the baseline measurement.
  • the increase is measured in a second tissue sample obtained from the subject after administering the composition to the subject.
  • the increase is measured directly in the subject after administering the composition to the subject.
  • the measurement is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline measurement.
  • the measurement is increased by about 10% or more, relative to the baseline measurement.
  • the measurement is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline measurement.
  • the measurement is increased by about 100% or more, increased by about 250% or more, increased by about 500% or more, increased by about 750% or more, or increased by about 1000% or more, relative to the baseline measurement. In some embodiments, the measurement is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline measurement. In some embodiments, the measurement is increased by no more than about 10%, relative to the baseline measurement. In some embodiments, the measurement is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline measurement.
  • the measurement is increased by no more than about 100%, increased by no more than about 250%, increased by no more than about 500%, increased by no more than about 750%, or increased by no more than about 1000%, relative to the baseline measurement. In some embodiments, the measurement is increased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the measurement is a questionnaire result.
  • the measurement is a hair loss assessment score.
  • the questionnaire result comprises a score such as a hair loss assessment score.
  • the questionnaire result is obtained from a questionnaire.
  • the questionnaire result is obtained from multiple questionnaires.
  • the questionnaire is a Men’s Hair Growth Questionnaire (MHGQ).
  • the questionnaire is a Kingsley Alopecia Profile (KAP) questionnaire.
  • the questionnaire may include questions about hair growth that the subject rates. Non-limiting examples of such ratings may include strongly agree, agree, neither agree nor disagree, disagree, and strongly disagree, where each rating is assigned a value.
  • the score may include a sum of each value.
  • the hair loss assessment score is not based on a questionnaire. In some embodiments, the hair loss assessment score is determined by a medical practitioner. In some embodiments, the hair loss assessment score includes a semi -quantitative hair visual hair score on a numerical scale such as 1-10. [00240] In some embodiments, the composition changes the hair loss assessment score relative to the baseline hair loss assessment score. In some embodiments, the change in the hair loss assessment score is an increase. In some embodiments, the change in the hair loss assessment score is a decrease. In some embodiments, the change is measured in the subject after administering the composition to the subject. In some embodiments, the change is measured directly by the subject after the composition is administered to the subject.
  • the hair loss assessment score is changed by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline hair loss assessment score. In some embodiments, the hair loss assessment score is changed by about 10% or more, relative to the baseline hair loss assessment score. In some embodiments, the hair loss assessment score is changed by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline hair loss assessment score. In some embodiments, the hair loss assessment score is changed by about 100% or more, changed by about 250% or more, changed by about 500% or more, changed by about 750% or more, or changed by about 1000% or more, relative to the baseline hair loss assessment score.
  • the hair loss assessment score is changed by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline hair loss assessment score. In some embodiments, the hair loss assessment score is changed by no more than about 10%, relative to the baseline hair loss assessment score. In some embodiments, the hair loss assessment score is changed by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline hair loss assessment score.
  • the hair loss assessment score is changed by no more than about 100%, changed by no more than about 250%, changed by no more than about 500%, changed by no more than about 750%, or changed by no more than about 1000%, relative to the baseline hair loss assessment score. In some embodiments, the hair loss assessment score is changed by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.
  • the hair loss assessment score is changed by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the measurement is a hair count.
  • the hair count is a total hair count.
  • the total hair count may include a vellus hair count and a non-vellus hair count.
  • the hair count is a vellus hair count.
  • the hair count is a non-vellus hair count.
  • the hair count is determined in an area of skin.
  • the hair count is normalized based on the area of skin.
  • the hair count is assessed using photography.
  • the hair count is assessed by phototrichogram.
  • the hair count is assessed by a macrophotography analysis.
  • the composition increases the hair count relative to the baseline hair count.
  • the increase is measured in the subject after administering the composition to the subject. In some embodiments, the increase is measured directly on the subject after administering the composition to the subject.
  • the hair count is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline hair count. In some embodiments, the hair count is increased by about 10% or more, relative to the baseline hair count. In some embodiments, the hair count is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline hair count.
  • the hair count is increased by about 100% or more, increased by about 250% or more, increased by about 500% or more, increased by about 750% or more, or increased by about 1000% or more, relative to the baseline hair count. In some embodiments, the hair count is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline hair count. In some embodiments, the hair count is increased by no more than about 10%, relative to the baseline hair count.
  • the hair count is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline hair count. In some embodiments, the hair count is increased by no more than about 100%, increased by no more than about 250%, increased by no more than about 500%, increased by no more than about 750%, or increased by no more than about 1000%, relative to the baseline hair count.
  • the hair count is increased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the measurement is a hair thickness measurement.
  • the hair thickness measurement is determined in an area of skin.
  • the hair thickness measurement comprises a width of an individual hair.
  • the hair thickness measurement comprises widths of multiple individual hairs.
  • the hair thickness measurement comprises an average of the widths of the multiple individual hairs.
  • the hair thickness measurement comprises a median of the widths of the multiple individual hairs.
  • the hair thickness measurement may include a vellus hair thickness measurement.
  • the hair thickness measurement may include a non -vellus hair thickness measurement.
  • the hair thickness measurement is assessed using photography.
  • the hair thickness measurement is assessed by phototrichogram.
  • the hair thickness measurement is assessed by a macrophotography analysis.
  • the composition increases the hair thickness measurement relative to the baseline hair thickness measurement.
  • the increase is measured in a second tissue sample (e.g. a skin sample as described herein) obtained from the subject after administering the composition to the subject.
  • the increase is measured directly on the subject after administering the composition to the subject.
  • the hair thickness measurement is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline hair thickness measurement.
  • the hair thickness measurement is increased by about 10% or more, relative to the baseline hair thickness measurement.
  • the hair thickness measurement is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline hair thickness measurement. In some embodiments, the hair thickness measurement is increased by about 100% or more, increased by about 250% or more, increased by about 500% or more, increased by about 750% or more, or increased by about 1000% or more, relative to the baseline hair thickness measurement. In some embodiments, the hair thickness measurement is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline hair thickness measurement. In some embodiments, the hair thickness measurement is increased by no more than about 10%, relative to the baseline hair thickness measurement.
  • the hair thickness measurement is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline hair thickness measurement. In some embodiments, the hair thickness measurement is increased by no more than about 100%, increased by no more than about 250%, increased by no more than about 500%, increased by no more than about 750%, or increased by no more than about 1000%, relative to the baseline hair thickness measurement.
  • the hair thickness measurement is increased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the measurement is a hair density measurement.
  • the hair density measurement is determined in an area of skin.
  • the hair density measurement comprises a number of hair in the area of skin.
  • the hair density measurement comprises the number of hair in the area of skin divided by the area of skin.
  • the hair density measurement may include a vellus hair density measurement.
  • the hair density measurement may include a non-vellus hair density measurement.
  • the hair density measurement is assessed using photography.
  • the hair density measurement is assessed by phototrichogram.
  • the hair density measurement is assessed by a macrophotography analysis.
  • the composition increases the hair density measurement relative to the baseline hair density measurement.
  • the increase is measured in a second tissue sample (e.g. a skin sample as described herein) obtained from the subject after administering the composition to the subject.
  • the increase is measured directly on the subject after administering the composition to the subject.
  • the hair density measurement is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline hair density measurement.
  • the hair density measurement is increased by about 10% or more, relative to the baseline hair density measurement.
  • the hair density measurement is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline hair density measurement. In some embodiments, the hair density measurement is increased by about 100% or more, increased by about 250% or more, increased by about 500% or more, increased by about 750% or more, or increased by about 1000% or more, relative to the baseline hair density measurement. In some embodiments, the hair density measurement is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline hair density measurement. In some embodiments, the hair density measurement is increased by no more than about 10%, relative to the baseline hair density measurement.
  • the hair density measurement is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline hair density measurement. In some embodiments, the hair density measurement is increased by no more than about 100%, increased by no more than about 250%, increased by no more than about 500%, increased by no more than about 750%, or increased by no more than about 1000%, relative to the baseline hair density measurement.
  • the hair density measurement is increased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the measurement is a number of hair follicles.
  • the number of hair follicles is a total number of hair follicles.
  • the number of hair follicles include a number of terminal hair follicles.
  • the number of hair follicles include a number of anagen hair follicles.
  • the number of hair follicles include a number of telogen hair follicles.
  • the number of hair follicles include a number of catagen hair follicles.
  • the number of hair follicles include a number of miniaturized hair follicles. In some embodiments, the number of hair follicles include a number of vellus miniaturized hair follicles. In some embodiments, the number of hair follicles include a number of vellus-like miniaturized hair follicles. In some embodiments, the number of hair follicles include a number of indeterminate hair follicles. In some embodiments, the number of hair follicles is determined in an area of skin. In some embodiments, the number of hair follicles is normalized based on the area of skin. In some embodiments, the number of hair follicles is assessed in a biopsy, such as a scalp biopsy. In some embodiments, the number of hair follicles is assessed using photography.
  • the composition increases the number of hair follicles relative to the baseline number of hair follicles.
  • the increase is measured in a second tissue sample (e.g. a skin sample as described herein) obtained from the subject after administering the composition to the subject.
  • the increase is measured directly on the subject after administering the composition to the subject.
  • the number of hair follicles is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline number of hair follicles.
  • the number of hair follicles is increased by about 10% or more, relative to the baseline number of hair follicles.
  • the number of hair follicles is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline number of hair follicles. In some embodiments, the number of hair follicles is increased by about 100% or more, increased by about 250% or more, increased by about 500% or more, increased by about 750% or more, or increased by about 1000% or more, relative to the baseline number of hair follicles. In some embodiments, the number of hair follicles is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline number of hair follicles.
  • the number of hair follicles is increased by no more than about 10%, relative to the baseline number of hair follicles. In some embodiments, the number of hair follicles is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline number of hair follicles.
  • the number of hair follicles is increased by no more than about 100%, increased by no more than about 250%, increased by no more than about 500%, increased by no more than about 750%, or increased by no more than about 1000%, relative to the baseline number of hair follicles. In some embodiments, the number of hair follicles is increased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the measurement is a protein level.
  • the protein level is a [3-catenin protein level.
  • the protein level is a a-SMA protein level.
  • the protein level is a collagen protein level.
  • the collagen of the collagen protein level is collagen I.
  • the collagen of the collagen protein level is collagen III.
  • the protein level is assessed in a sample such as a skin sample.
  • the protein level is indicated as a mass or percentage of protein per sample weight.
  • the protein level is indicated as a mass or percentage of protein per sample volume.
  • the protein level is indicated as a mass or percentage of protein per total protein within the sample.
  • the protein measurement is a circulating protein measurement.
  • the protein measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.
  • the measurement is a hair color measurement.
  • the hair color measurement is determined in an area of skin.
  • the hair color measurement comprises a color of an individual hair.
  • the hair color measurement comprises colors of multiple individual hairs.
  • the hair color measurement comprises an average of the colors of the multiple individual hairs.
  • the hair color measurement comprises a median of the colors of the multiple individual hairs.
  • the hair color measurement may include a measurement of how gray the hair is, or how gray the hairs are.
  • the hair color measurement may include a measurement of how much color is in the hair, or how much color is in the hairs.
  • the hair color measurement may include a measurement of how white the hair is, or how white the hairs are.
  • the hair color measurement may include a hair pigmentation measurement.
  • the hair color measurement may include a hair contrast measurement.
  • the hair color measurement may include a vellus hair color measurement.
  • the hair color measurement may include a non-vellus hair color measurement.
  • the hair color measurement is assessed using photography.
  • the hair color measurement is assessed by phototrichogram.
  • the hair color measurement is assessed by a macrophotography analysis.
  • the hair color measurement may be a qualitative measurement.
  • the hair color measurement may be a quantitative measurement.
  • the hair color measurement may be a number, such as an amount of hair color.
  • the hair color measurement may be a rate, such as a rate of hair color loss.
  • the composition increases the hair color measurement relative to the baseline hair color measurement.
  • the increase is measured in a second tissue sample (e.g. a skin sample as described herein) obtained from the subject after administering the composition to the subject.
  • the increase is measured directly on the subject after administering the composition to the subject.
  • the hair color measurement is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline hair color measurement.
  • the hair color measurement is increased by about 10% or more, relative to the baseline hair color measurement.
  • the hair color measurement is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline hair color measurement. In some embodiments, the hair color measurement is increased by about 100% or more, increased by about 250% or more, increased by about 500% or more, increased by about 750% or more, or increased by about 1000% or more, relative to the baseline hair color measurement. In some embodiments, the hair color measurement is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline hair color measurement. In some embodiments, the hair color measurement is increased by no more than about 10%, relative to the baseline hair color measurement.
  • the hair color measurement is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline hair color measurement. In some embodiments, the hair color measurement is increased by no more than about 100%, increased by no more than about 250%, increased by no more than about 500%, increased by no more than about 750%, or increased by no more than about 1000%, relative to the baseline hair color measurement.
  • the hair color measurement is increased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the composition reduces the protein level relative to the baseline protein level.
  • the reduction is measured in a second tissue or fluid sample (e.g. a skin, blood, serum, or plasma sample as described herein) obtained from the subject after administering the composition to the subject.
  • the protein level is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline protein level. In some embodiments, the protein level is decreased by about 10% or more, relative to the baseline protein level.
  • the protein level is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline protein level. In some embodiments, the protein level is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline protein level. In some embodiments, the protein level is decreased by no more than about 10%, relative to the baseline protein level.
  • the protein level is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline protein level. In some embodiments, the protein level is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.
  • the composition increases the protein level relative to the baseline protein level.
  • the increase is measured in a second tissue or fluid sample (e.g. a skin, blood, serum, or plasma sample as described herein) obtained from the subject after administering the composition to the subject.
  • the protein level is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline protein level.
  • the protein level is increased by about 10% or more, relative to the baseline protein level.
  • the protein level is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline protein level. In some embodiments, the protein level is increased by about 100% or more, increased by about 250% or more, increased by about 500% or more, increased by about 750% or more, or increased by about 1000% or more, relative to the baseline protein level. In some embodiments, the protein level is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline protein level. In some embodiments, the protein level is increased by no more than about 10%, relative to the baseline protein level.
  • the protein level is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline protein level. In some embodiments, the protein level is increased by no more than about 100%, increased by no more than about 250%, increased by no more than about 500%, increased by no more than about 750%, or increased by no more than about 1000%, relative to the baseline protein level.
  • the protein level is increased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the measurement is a mRNA level.
  • the mRNA level is a [3-catenin mRNA level.
  • the mRNA level is a a-SMA mRNA level.
  • the mRNA level is a collagen mRNA level.
  • the collagen of the collagen mRNA level is collagen I.
  • the collagen of the collagen mRNA level is collagen III.
  • the mRNA level is assessed in a sample such as a skin sample. In some embodiments, the mRNA level is indicated as a mass or percentage of mRNA per sample weight.
  • the mRNA level is indicated as a mass or percentage of mRNA per sample volume. In some embodiments, the mRNA level is indicated as a mass or percentage of mRNA per total mRNA within the sample. In some embodiments, the mRNA level is indicated as a mass or percentage of mRNA per total nucleic acids within the sample. In some embodiments, the mRNA level is indicated relative to another mRNA level, such as an mRNA level of a housekeeping gene, within the sample. In some embodiments, the mRNA measurement is obtained by an assay such as a polymerase chain reaction (PCR) assay. In some embodiments, the PCR comprises quantitative PCR (qPCR). In some embodiments, the PCR comprises reverse transcription of the mRNA.
  • PCR polymerase chain reaction
  • the composition reduces the mRNA level relative to the baseline mRNA level.
  • the reduction is measured in a second tissue sample (e.g. a skin sample as described herein) obtained from the subject after administering the composition to the subject.
  • the mRNA level is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline mRNA level.
  • the mRNA level is decreased by about 10% or more, relative to the baseline mRNA level.
  • the mRNA level is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline mRNA level. In some embodiments, the mRNA level is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline mRNA level. In some embodiments, the mRNA level is decreased by no more than about 10%, relative to the baseline mRNA level.
  • the mRNA level is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline mRNA level. In some embodiments, the mRNA level is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.
  • the composition increases the mRNA level relative to the baseline mRNA level.
  • the increase is measured in a second tissue sample (e.g. a skin sample as described herein) obtained from the subject after administering the composition to the subject.
  • the mRNA level is increased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline mRNA level.
  • the mRNA level is increased by about 10% or more, relative to the baseline mRNA level.
  • the mRNA level is increased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline mRNA level. In some embodiments, the mRNA level is increased by about 100% or more, increased by about 250% or more, increased by about 500% or more, increased by about 750% or more, or increased by about 1000% or more, relative to the baseline mRNA level. In some embodiments, the mRNA level is increased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline mRNA level.
  • the mRNA level is increased by no more than about 10%, relative to the baseline mRNA level. In some embodiments, the mRNA level is increased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline mRNA level. In some embodiments, the mRNA level is increased by no more than about 100%, increased by no more than about 250%, increased by no more than about 500%, increased by no more than about 750%, or increased by no more than about 1000%, relative to the baseline mRNA level.
  • the mRNA level is increased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 250%, 500%, 750%, or 1000%, or by a range defined by any of the two aforementioned percentages.
  • the measurement is a DKK2 protein measurement.
  • the DKK2 protein measurement comprises a DKK2 protein level.
  • the DKK2 protein level is assessed in a sample such as a skin sample or a fluid sample (e.g. blood, serum, or plasma).
  • the DKK2 protein level is indicated as a mass or percentage of DKK2 protein per sample weight.
  • the DKK2 protein level is indicated as a mass or percentage of DKK2 protein per sample volume.
  • the DKK2 protein level is indicated as a mass or percentage of DKK2 protein per total protein within the sample.
  • the DKK2 protein measurement is a circulating DKK2 protein measurement.
  • the DKK2 protein measurement is obtained by an assay such as an immunoassay, a colorimetric assay, or a fluorescence assay.
  • the composition reduces the DKK2 protein level relative to the baseline DKK2 protein level.
  • the reduction is measured in a second tissue or fluid sample (e.g. a skin, blood, serum, or plasma sample as described herein) obtained from the subject after administering the composition to the subject.
  • the DKK2 protein level is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline DKK2 protein level.
  • the DKK2 protein level is decreased by about 10% or more, relative to the baseline DKK2 protein level.
  • the DKK2 protein level is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline DKK2 protein level. In some embodiments, the DKK2 protein level is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline DKK2 protein level. In some embodiments, the DKK2 protein level is decreased by no more than about 10%, relative to the baseline DKK2 protein level.
  • the DKK2 protein level is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline DKK2 protein level. In some embodiments, the DKK2 protein level is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.
  • the measurement is a DKK2 mRNA measurement.
  • the DKK2 mRNA measurement comprises a DKK2 mRNA level.
  • the DKK2 mRNA level is assessed in a sample such as a skin sample.
  • the DKK2 mRNA level is indicated as a mass or percentage of DKK2 mRNA per sample weight.
  • the DKK2 mRNA level is indicated as a mass or percentage of DKK2 mRNA per sample volume.
  • the DKK2 mRNA level is indicated as a mass or percentage of DKK2 mRNA per total mRNA within the sample.
  • the DKK2 mRNA level is indicated as a mass or percentage of DKK2 mRNA per total nucleic acids within the sample. In some embodiments, the DKK2 mRNA level is indicated relative to another mRNA level, such as an mRNA level of a housekeeping gene, within the sample. In some embodiments, the DKK2 mRNA measurement is obtained by an assay such as a polymerase chain reaction (PCR) assay. In some embodiments, the PCR comprises quantitative PCR (qPCR). In some embodiments, the PCR comprises reverse transcription of the DKK2 mRNA.
  • PCR polymerase chain reaction
  • the composition reduces the DKK2 mRNA level relative to the baseline DKK2 mRNA level.
  • the reduction is measured in a second tissue sample (e.g. a skin sample as described herein) obtained from the subject after administering the composition to the subject.
  • the DKK2 mRNA level is decreased by about 2.5% or more, about 5% or more, or about 7.5% or more, relative to the baseline DKK2 mRNA level.
  • the DKK2 mRNA level is decreased by about 10% or more, relative to the baseline DKK2 mRNA level.
  • the DKK2 mRNA level is decreased by about 20% or more, about 30% or more, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, relative to the baseline DKK2 mRNA level. In some embodiments, the DKK2 mRNA level is decreased by no more than about 2.5%, no more than about 5%, or no more than about 7.5%, relative to the baseline DKK2 mRNA level. In some embodiments, the DKK2 mRNA level is decreased by no more than about 10%, relative to the baseline DKK2 mRNA level.
  • the DKK2 mRNA level is decreased by no more than about 20%, no more than about 30%, no more than about 40%, no more than about 50%, no more than about 60%, no more than about 70%, no more than about 80%, no more than about 90%, or no more than about 100% relative to the baseline DKK2 mRNA level. In some embodiments, the DKK2 mRNA level is decreased by 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%, or by a range defined by any of the two aforementioned percentages.
  • determining means determining if an element is present or not (for example, detection). These terms can include quantitative, qualitative or quantitative and qualitative determinations. Assessing can be relative or absolute. “Detecting the presence of’ can include determining the amount of something present in addition to determining whether it is present or absent depending on the context.
  • a “subject” can be a biological entity containing expressed genetic materials.
  • the biological entity can be a plant, animal, or microorganism, including, for example, bacteria, viruses, fungi, and protozoa.
  • the subject can be a mammal.
  • the mammal can be a human.
  • the subject may be diagnosed or suspected of being at high risk for a disease. In some cases, the subject is not necessarily diagnosed or suspected of being at high risk for the disease.
  • the term “about” a number refers to that number plus or minus 10% of that number.
  • the term “about” a range refers to that range minus 10% of its lowest value and plus 10% of its greatest value.
  • treatment or “treating” are used in reference to a pharmaceutical or other intervention regimen for obtaining beneficial or desired results in the recipient.
  • beneficial or desired results include but are not limited to a therapeutic benefit and/or a prophylactic benefit.
  • a therapeutic benefit may refer to eradication or amelioration of symptoms or of an underlying disorder being treated.
  • a therapeutic benefit can be achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder.
  • a prophylactic effect includes delaying, preventing, or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof.
  • a subject at risk of developing a particular disease, or to a subject reporting one or more of the physiological symptoms of a disease may undergo treatment, even though a diagnosis of this disease may not have been made.
  • any uracil (U) may be interchanged with any thymine (T), and vice versa.
  • any of the Us may be replaced with Ts.
  • an siRNA with a nucleic acid sequence comprising one or more Ts in some embodiments any of the Ts may be replaced with Us.
  • an oligonucleotide such as an siRNA disclosed herein comprises or consists of RNA.
  • the oligonucleotide may comprise or consist of DNA.
  • C x.y or “C x -C y ” when used in conjunction with a chemical moiety, such as alkyl, alkenyl, or alkynyl is meant to include groups that contain from x to y carbons in the chain.
  • Ci-ealkyl refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups that contain from 1 to 6 carbons.
  • C x.y alkenyl and C x.y alkynyl refer to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively.
  • Carbocycle refers to a saturated, unsaturated or aromatic ring in which each atom of the ring is carbon.
  • Carbocycle includes 3- to 10-membered monocyclic rings, 5- to 12- membered bicyclic rings, 5- to 12-membered spiro bicycles, and 5- to 12-membered bridged rings.
  • Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated, and aromatic rings.
  • an aromatic ring e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene.
  • a bicyclic carbocycle includes any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits.
  • a bicyclic carbocycle further includes spiro bicyclic rings such as spiropentane.
  • a bicyclic carbocycle includes any combination of ring sizes such as 3-3 spiro ring systems, 4-4 spiro ring systems, 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5- 8 fused ring systems, and 6-8 fused ring systems.
  • Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, naphthyl, and bicyclo [l.l.l]pentanyl.
  • aryl refers to an aromatic monocyclic or aromatic multicyclic hydrocarbon ring system.
  • the aromatic monocyclic or aromatic multicyclic hydrocarbon ring system contains only hydrogen and carbon and from five to eighteen carbon atoms, where at least one of the rings in the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) n-electron system in accordance with the Htickel theory.
  • the ring system from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin and naphthalene.
  • cycloalkyl refers to a saturated ring in which each atom of the ring is carbon.
  • Cycloalkyl may include monocyclic and polycyclic rings such as 3- to 10-membered monocyclic rings, 5- to 12-membered bicyclic rings, 5- to 12-membered spiro bicycles, and 5- to 12-membered bridged rings.
  • a cycloalkyl comprises three to ten carbon atoms.
  • a cycloalkyl comprises five to seven carbon atoms.
  • the cycloalkyl may be attached to the rest of the molecule by a single bond.
  • Examples of monocyclic cycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • Polycyclic cycloalkyl radicals include, for example, adamantyl, spiropentane, norbomyl (i.e., bicyclo[2.2.1]heptanyl), decalinyl, 7,7 dimethyl bicyclo[2.2.1]heptanyl, bicyclo [l. l.l]pentanyl, and the like.
  • cycloalkenyl refers to a saturated ring in which each atom of the ring is carbon and there is at least one double bond between two ring carbons.
  • Cycloalkenyl may include monocyclic and polycyclic rings such as 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 5- to 12-membered bridged rings.
  • a cycloalkenyl comprises five to seven carbon atoms.
  • the cycloalkenyl may be attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.
  • halo or, alternatively, “halogen” or “halide,” means fluoro, chloro, bromo or iodo. In some embodiments, halo is fluoro, chloro, or bromo.
  • haloalkyl refers to an alkyl radical, as defined above, that is substituted by one or more halo radicals, for example, trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1- chloromethyl -2 -fluoroethyl, and the like.
  • the alkyl part of the haloalkyl radical is optionally further substituted as described herein.
  • heterocycle refers to a saturated, unsaturated or aromatic ring comprising one or more heteroatoms.
  • exemplary heteroatoms include N, O, Si, P, B, and S atoms.
  • Heterocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, 5- to 12- membered spiro bicycles, and 5- to 12-membered bridged rings.
  • a bicyclic heterocycle includes any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits.
  • an aromatic ring e.g., pyridyl
  • a saturated or unsaturated ring e.g., cyclohexane, cyclopentane, morpholine, piperidine or cyclohexene.
  • a bicyclic heterocycle includes any combination of ring sizes such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems.
  • a bicyclic heterocycle further includes spiro bicyclic rings, e.g., 5 to 12-membered spiro bicycles, such as 2-oxa-6-azaspiro[3.3]heptane.
  • heteroaryl refers to a radical derived from a 5 to 18 membered aromatic ring radical that comprises two to seventeen carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur.
  • the heteroaryl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, wherein at least one of the rings in the ring system is aromatic, i.e., it contains a cyclic, delocalized (4n+2) n-electron system in accordance with the Htickel theory.
  • Heteroaryl includes fused or bridged ring systems.
  • the heteroatom(s) in the heteroaryl radical is optionally oxidized.
  • heteroaryl is attached to the rest of the molecule through any atom of the ring(s).
  • heteroaryls include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzoxazolyl, benzo [d]thiazolyl, benzothiadiazolyl, benzo[b][l,4]dioxepinyl, benzo[b][l,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzo
  • heterocycloalkyl refers to a saturated ring with carbon atoms and at least one heteroatom.
  • exemplary heteroatoms include N, O, Si, P, B, and S atoms.
  • Heterocycloalkyl may include monocyclic and polycyclic rings such as 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, 5- to 12-membered spiro bicycles, and 5- to 12-membered bridged rings.
  • the heteroatoms in the heterocycloalkyl radical are optionally oxidized.
  • One or more nitrogen atoms, if present, are optionally quatemized.
  • heterocycloalkyl is attached to the rest of the molecule through any atom of the heterocycloalkyl, valence permitting, such as any carbon or nitrogen atoms of the heterocycloalkyl.
  • heterocycloalkyl radicals include, but are not limited to, dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,
  • heterocycloalkenyl refers to an unsaturated ring with carbon atoms and at least one heteroatom and there is at least one double bond between two ring carbons. Heterocycloalkenyl does not include heteroaryl rings. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycloalkenyl may include monocyclic and polycyclic rings such as 3- to 10-membered monocyclic rings, 6- to 12- membered bicyclic rings, and 5- to 12-membered bridged rings. In other embodiments, a heterocycloalkenyl comprises five to seven ring atoms.
  • the heterocycloalkenyl may be attached to the rest of the molecule by a single bond.
  • monocyclic cycloalkenyls include, e.g., pyrroline (dihydropyrrole), pyrazoline (dihydropyrazole), imidazoline (dihydroimidazole), triazoline (dihydrotriazole), dihydrofuran, dihydrothiophene, oxazoline (dihydrooxazole), isoxazoline (dihydroisoxazole), thiazoline (dihydrothiazole), isothiazoline (dihydroisothiazole), oxadiazoline (dihydrooxadiazole), thiadiazoline (dihydrothiadiazole), dihydropyridine, tetrahydropyridine, dihydropyridazine, tetrahydropyridazine, dihydropyrimidine, tetrahydro
  • substituted refers to moieties having substituents replacing a hydrogen on one or more carbons or substitutable heteroatoms, e.g., an NH or NH2 of a compound. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, i.e., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.
  • substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as substituting the two hydrogen atoms on a single carbon with an oxo, imino or thioxo group.
  • substituted is contemplated to include all permissible substituents of organic compounds.
  • the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non -aromatic substituents of organic compounds.
  • the permissible substituents can be one or more and the same or different for appropriate organic compounds.
  • a "derivative" polypeptide or peptide is one that is modified, for example, by glycosylation, pegylation, phosphorylation, sulfation, reduction/alkylation, acylation, chemical coupling, or mild formalin treatment.
  • a derivative may also be modified to contain a detectable label, either directly or indirectly, including, but not limited to, a radioisotope, fluorescent, and enzyme label.
  • Some embodiments refer to nucleic acid sequence information. It is contemplated that in some embodiments, thymine (T) may be interchanged with uracil (U), or vice versa.
  • sequences in the sequence listing may recite Ts, but these may be replaced with Us in some embodiments.
  • the uracil may be replaced with thymine.
  • the thymine may be replaced with uracil.
  • an oligonucleotide such as an siRNA comprises or consists of RNA.
  • the oligonucleotide may include DNA.
  • the oligonucleotide may include 2’ deoxyribonucleotides.
  • An ASO may comprise or consist of DNA.
  • Nf e.g. Af, Cf, Gf, Tf, or Uf
  • dN e.g. dA, dC, dG, dT, or dU
  • n e.g. a, c, g, t, or u
  • s refers to a phosphorothioate linkage.
  • a pyrimidine may include cytosine (C), thymine (T), or uracil (U).
  • a pyrimidine may include C or U.
  • a pyrimidine may include C or T. Where a pyrimidine is referred to, it may indicate a nucleoside or nucleotide comprising a pyrimidine.
  • a purine may include guanine (G), inosine (I), adenine (A). Where a purine is referred to, it may indicate a nucleoside or nucleotide comprising a purine.
  • Some embodiments include one or more nucleic acid sequences in the following table:
  • Example 1 Variants in DKK2 are associated with decreased risk of male pattern baldness
  • Variants in DKK2 were evaluated for associations with male pattern baldness (MBP) in approximately 206,000 male individuals with genotype data from the UK Biobank cohort.
  • the two variants were considered to be hypomorphic or loss of function variants that may result in a decrease in the abundance or activity of the DKK2 gene product.
  • Lymphoblastoid cell lines from three age and gender-matched donors with known rs35290077 (G96R) genotypes, including a donor that was homozygous for the reference allele (rs35290077 C/C), a donor that was heterozygous (rs35290077 C/G) and a donor that was homozygous for alternative allele (rs35290077 G/G), were seeded at 500,000 cells/well in 24 well plate in complete growth media and grown overnight. LCLs were treated with 25 nM Vitamin D or vehicle (100% EtOH) for 2 days, and then harvested.
  • DKK2 gene variants associated with protection from MBP result in loss of DKK2 mRNA abundance or function. Accordingly, in some cases therapeutic inhibition or modulation of DKK2 may be an effective genetically -informed method of treatment for MBP and related traits or diseases.
  • Example 2 Bioinformatic selection of sequences in order to identify therapeutic siRNAs to downmodulate expression of the DKK2 mRNA
  • siRNAs were designed to target human DKK2. Predicted specificity in human, rhesus monkey, cynomolgus monkey, mouse, rat, rabbit, and dog was determined for sense (S) and antisense (AS) strands. These were assigned a “specificity score” which considers the likelihood of unintended downregulation of any other transcript by full or partial complementarity of an siRNA strand (up to 2 mismatches within positions 2-18) as well as the number and positions of mismatches. Thus, off-target(s) transcripts for antisense and sense strands of each siRNA were identified. As identified, siRNAs with high specificity and a low number of predicted off-targets provided a benefit of increased targeting specificity.
  • siRNA sequences within the seed region were analyzed for similarity to seed regions of known miRNAs.
  • siRNAs can function in a miRNA like manner via base-pairing with complementary sequences within the 3’-UTR of mRNA molecules. The complementarity typically encompasses the 5‘-bases at positions 2-7 of the miRNA (seed region).
  • siRNA strands containing natural miRNA seed regions can be avoided. Seed regions identified in miRNAs from human, mouse, rat, rhesus monkey, dog, rabbit, and pig are referred to as “conserved”. Combining the “specificity score” with miRNA seed analysis yielded a “specificity category”. This is divided into categories 1-4, with 1 having the highest specificity and 4 having the lowest specificity. Each strand of the siRNA is assigned to a specificity category.
  • the siRNAs in these subsets recognized at least the human DKK2 sequences. Therefore, the siRNAs in these subsets can be used to target human DKK2 in a therapeutic setting.
  • siRNA sequences derived from human DKK2 mRNA (ENST00000285311, SEQ ID NO: 7599) without consideration of specificity or species cross-reactivity was 3636 (sense and antisense strand sequences included in SEQ ID NOS : 1-3636 and 3637-7272, respectively).
  • Subset A contained 741 siRNAs, including siRNAs 12, 13, 21, 22, 24, 27, 29, 30, 31, 34, 35, 36, 38, 39, 40, 42, 43, 46, 47, 48, 51, 53, 54, 60, 63, 64, 65, 66, 67, 68, 69, 70, 74, 76, 77, 81, 84, 93, 94, 95, 98, 102, 103, 104, 105, 107, 117, 118, 119, 122, 124, 130, 131, 132, 135, 136, 137, 142, 144, 145, 149, 150, 156, 157, 159, 160, 161, 164, 165, 166, 167, 168, 169, 170, 175, 179,
  • siRNAs in subset A had the following characteristics: Cross-reactivity: With 19mer in human DKK2 mRNA; Specificity category: For human: AS2 or better, SS3 or better; and miRNA seeds: AS+SS strand: seed region not conserved in human, mouse, and rat and not present in >4 species; Off- target frequency: ⁇ 30 human off-targets matched with 2 mismatches in antisense strand; and SNPs: siRNA target sites do not harbor SNPs with a MAF > 1% (pos. 2-18).
  • subset A The siRNA sequences in subset A were selected for more stringent specificity to yield subset B.
  • Subset B included 735 siRNAs, including siRNAs 12, 13, 21, 22, 24, 27, 29, 30, 31, 34, 35, 36, 38, 39, 40, 42, 43, 46, 47, 48, 51, 53, 54, 60, 63, 64, 65, 66, 67, 68, 69, 70, 74, 76, 77, 81, 84, 93, 94, 95, 98, 102, 103, 104, 105, 107, 117, 118, 119, 124, 130, 131, 132, 135, 136, 137, 142, 144, 145, 149, 150, 156, 157,
  • siRNAs in subset B had the following characteristics: Cross-reactivity: With 19mer in human DKK2 mRNA; Specificity category: For human: AS2 or better, SS3 or better; miRNA seeds: AS+SS strand: seed region not conserved in human, mouse, and rat and not present in >4 species; Off-target frequency: ⁇ 20 human off-targets matched with 2 mismatches in antisense strand; and SNPs: siRNA target sites do not harbor SNPs with a MAF > 1% (pos. 2-18).
  • subset C includes 489 siRNAs, including siRNAs 12, 13, 21, 22, 24, 27, 29, 30, 31, 35, 36, 40, 43, 46, 47, 48, 51, 53, 54, 60, 63, 64, 65, 66, 67, 68, 69, 74, 76, 81, 84, 93, 94, 95, 102, 103, 105, 107, 119, 122, 130, 131, 132, 135, 136, 142, 144, 149, 150, 156, 157, 159, 160, 165, 166, 167, 168, 170, 175, 184, 186, 188, 190, 191, 192,
  • siRNAs in subset C had the following characteristics: Cross-reactivity: With 19mer in human DKK2 mRNA; Specificity category: For human: AS2 or better, SS3 or better; miRNA seeds: AS+SS strand: seed region not conserved in human, mouse, and rat and not present in >4 species. AS strand: seed region not identical to seed region of known human miRNA; Off-target frequency: ⁇ 30 human off-targets matched with 2 mismatches by antisense strand; and SNPs: siRNA target sites do not harbor SNPs with a MAF > 1% (pos. 2-18).
  • subset C The siRNA sequences in subset C were also selected for absence of seed regions in the AS or S strands that are identical to a seed region of known human miRNA in addition to having an off-target frequency of ⁇ 30 human off-targets matched with 2 mismatches by antisense strand to yield subset D.
  • Subset D includes 311 siRNAs, including siRNAs 12, 24, 29, 30, 31, 35, 36, 40, 43, 46, 47, 51, 64, 65, 66, 74, 76, 94, 95, 103, 105, 122, 130, 131, 142, 144, 160, 166, 167, 168, 186, 190, 192, 220, 229, 233, 241,
  • subset D The siRNA sequences in subset D were also selected to have an off-target frequency of ⁇ 20 human off-targets matched with 2 mismatches by antisense strand to yield subset E.
  • Subset E includes 307 siRNAs, including siRNAs 12, 24, 29, 30, 31, 35, 36, 40, 43, 46, 47, 51, 64, 65, 66, 74, 76, 94, 95, 103, 105, 130, 131, 142, 144, 160, 166, 167, 168, 186, 190, 192, 220, 229, 233, 242, 243, 283, 308, 324, 338,
  • siRNAs were designed to target human DKK2 as described above and, in some cases, the DKK2 sequence of at least one toxicology-relevant species, in this case, the non-human primate (NHP) cynomolgus monkey.
  • the siRNAs included in subset F had the following characteristics: Crossreactivity: With 19mer in human DKK2 mRNA, with 17mer/19mer in NHP DKK2; Specificity category: For human and NHP: AS2 or better, SS3 or better.
  • Subset F includes 40 siRNAs, including siRNAs 822, 824, 827, 918, 949, 950, 951, 1083, 1180, 1182, 1203, 1214, 1218, 1219, 1221, 1223, 1227, 1229, 1236, 1292, 1319, 1320, 1325, 1328, 1443, 1446, 1550, 2348, 2588, 2589, 2590, 3508, 3525, 3545, 3547, 3555, 3556, 3564, 3572, and 3579.
  • the sense strand of any of the siRNAs of subset F comprises siRNA with a particular modification pattern.
  • position 9 counting from the 5’ end of the of the sense strand is has the 2’F modification.
  • position 9 of the sense strand is a pyrimidine, then all purines in the sense strand have the 2’OMe modification.
  • position 9 is the only pyrimidine between positions 5 and 11 of the sense stand, then position 9 is the only position with the 2’F modification in the sense strand.
  • position 9 and only one other base between positions 5 and 11 of the sense strand are pyrimidines, then both of these pyrimidines are the only two positions with the 2’F modification in the sense strand.
  • position 9 is the only purine between positions 5 and 11 of the sense stand, then position 9 is the only position with the 2’F modification in the sense strand. If position 9 and only one other base between positions 5 and 11 of the sense strand are purines, then both of these purines are the only two positions with the 2’F modification in the sense strand. If position 9 and only two other bases between positions 5 and 11 of the sense strand are purines, and those two other purines are in adjacent positions so that there would be not three 2’F modifications in a row, then any combination of 2’F modifications can be made that give three 2’F modifications in total.
  • position 9 of the sense strand can be a 2’deoxy. In these cases, 2’F and 2’0Me modifications may occur at the other positions of the sense strand.
  • the sense strand of any of the siRNAs of subset F comprises a modification pattern which conforms to these sense strand rules (Table 4).
  • the antisense strand of any of the siRNAs of subset F comprise a modification or modification pattern. Some such examples are included in Table 4. Table 5 includes some additional sense strand modifications of the siRNAs in subset F. The siRNAs in subset F may comprise any other modification pattem(s).
  • Nf e.g. Af, Cf, Gf, Tf, or Uf
  • dN e.g. dA, dC, dG, dT, or dU
  • n e.g. a, c, g, t, or u
  • s is a phosphorothioate linkage.
  • any siRNA among any of subsets A-H may comprise any modification pattern described herein. If a sequence has a different number of nucleotides in length than a modification pattern, the modification pattern may still be used with the appropriate number of additional nucleotides added 5 ’ or 3 ’ to match the number of nucleotides in the modification pattern. For example, if a sense or antisense strand of the siRNA among any of subsets A-H comprises 19 nucleotides, and a modification pattern comprises 21 nucleotides, UU may be added onto the 5’ end of the sense or antisense strand.
  • Example 3 In vivo delivery of siRNA in C57 mice
  • ETD01043 or ETD01551 formulated in 10 pl of 1 part azone 32 parts propylene glycol was applied to the shaved area.
  • ETD01043 was used as a control and targeted human ANGPTL7, and ETD01551 targeted DKK2.
  • the application of siRNA’s was repeated on days, 2, 5 and 7.
  • formulations containing 30 ug ETD01043 or ETD01551, and 10 ug dexamethasone acetate formulated in 10 pl of 1 part azone: 32 parts propylene glycol were applied to the shaved areas.
  • the formulations containing dexamethasone and siRNA were repeated on days 10, 11, 12, 13 and 14.
  • RNA samples of the siRNA-applied areas were obtained by punch biopsy and placed into RNAlater.
  • Total skin RNA was prepared by homogenizing the tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles. The homogenate was centrifuged for 10’ at 16,000xg at 4C and the lower liquid layer was removed to a fresh tube. The sample was centrifuged two additional times, each time removing the lower liquid layer to a fresh tube.
  • RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations.
  • Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions.
  • the relative levels of mouse DKK2 mRNA were assessed by RT-qPCR in triplicate on a QuantStudioTM 6 Pro Real-Time PCR System using and the mouse housekeeping gene PPIA (ThermoFisher, assay#Mm02342430_g I ). Data were normalized to the level in animals receiving ETD01043, which showed that an average DKK2 knockdown of 87% for the animals treated with DKK2- targeting ETD01551 .
  • siRNAs are shown in Table 6, where Nf (e.g. Af, Cf, Gf, Tf, or Uf) is a 2’-fluoro- modified nucleoside, n (e.g. a, c, g, t, or u) is a 2’-O-methyl modified nucleoside, and “s” is a phosphorothioate linkage.
  • Nf e.g. Af, Cf, Gf, Tf, or Uf
  • n e.g. a, c, g, t, or u
  • siRNAs in this table may target mouse DKK2. Table 6.
  • Example 4 siRNA-mediated knockdown of DKK2 in dermal fibroblast cell line
  • siRNAs targeted to the DKK2 mRNA that downregulate levels of DKK2 mRNA are transfected into cultured dermal fibroblast cell. Downregulation of DKK2 mRNA (and ultimately protein) subsequently leads to an increase in protein levels of [3-catenin, a-SMA, and collagens I and III in cultured dermal fibroblast cells.
  • the dermal fibroblast cells are seeded at 150,000 cells/mL into a Falcon 24-well tissue culture plate (Cat. No. 353047) at 0.5 mL per well.
  • the DKK2 siRNA and negative control siRNA master mixes are prepared.
  • the DKK2 siRNA master mix contains 350 pL of Opti-MEM (ThermoFisher Cat. No. 4427037 - sl288 Lot No. AS02B02D) and 3.5 pL of a mixture of the two DKK2 siRNAs (10 pM stock).
  • the negative control siRNA master mix contains 350 pL of Opti-MEM and 3.5 pL of negative control siRNA (ThermoFisher Cat. No. 4390843, 10 pM stock).
  • 3 pL of TransIT-X2 (Minis Cat. No. MIR6000) is added to each master mix.
  • the mixes are incubated for 15 minutes to allow transfection complexes to form, then 51 pL of the appropriate master mix + TransIT-X2 is added to quadruplicate wells of dermal fibroblast cells with a final siRNA concentration of 10 nM.
  • SDS-PAGE is done in 8% glycine gels (Bio-rad) loading equal amount of proteins per lane. After electrophoresis, separated proteins are transferred to nitrocellulose membrane (Bio-rad) and blocked with 5% non-fat milk in TBST buffer for Ih.
  • the membranes are incubated with DKK2 (1:600; CST), [3-Catenin (1:800; CST), alpha smooth muscle Actin (1:800; abeam), Collagen I (1:800; abeam), Collagen III (1:800; abeam) and GAPDH (1:2,000; CST), loading control, antibodies overnight at 4°C, and then anti-rabbit IgG monoclonal antibody conjugated with horseradish peroxidase (Pierce) at 1:2000 dilution for 1 h at room temperature. Protein bands are detected using the West Femto system (Pierce).
  • a decrease in DKK2 mRNA expression in the dermal fibroblast cells is expected after transfection with the DKK2 siRNAs compared to DKK2 mRNA levels in dermal fibroblast cells transfected with the non-specific control siRNA 48 hours after transfection.
  • Example 5 ASO-mediated knockdown of DKK2 in dermal fibroblast cell line
  • ASOs targeted to the DKK2 mRNA that downregulate levels of DKK2 mRNA are transfected into cultured dermal fibroblast cell. Downregulation of DKK2 mRNA (and ultimately protein) subsequently leads to an increase in protein levels of [3-catenin, a-SMA, and collagens I and III in cultured dermal fibroblast cells.
  • the dermal fibroblast cells are seeded at 150,000 cells/mL into a Falcon 24-well tissue culture plate (Cat. No. 353047) at 0.5 mb per well.
  • the DKK2 ASO and negative control ASO master mixes are prepared.
  • the DKK2 ASO master mix contains 350 pL of Opti-MEM (ThermoFisher Cat. No. 4427037 - sl288 Lot No. AS02B02D) and 3.5 pL of a mixture of the two DKK2 ASOs (10 pM stock).
  • the negative control ASO master mix contains 350 pL of Opti-MEM and 3.5 pL of negative control ASO (ThermoFisher Cat. No. 4390843, 10 pM stock).
  • 3 pL of TransIT-X2 (Minis Cat. No. MIR6000) is added to each master mix.
  • the mixes are incubated for 15 minutes to allow transfection complexes to form, then 51 pL of the appropriate master mix + TransIT-X2 is added to quadruplicate wells of dermal fibroblast cells with a final ASO concentration of 10 nM.
  • SDS-PAGE is done in 8% glycine gels (Bio-rad) loading equal amount of proteins per lane. After electrophoresis, separated proteins are transferred to nitrocellulose membrane (Bio-rad) and blocked with 5% non-fat milk in TBST buffer for Ih.
  • the membranes are incubated with DKK2 (1:600; CST), [3-Catenin (1:800; CST), alpha smooth muscle Actin (1:800; abeam), Collagen I (1:800; abeam), Collagen III (1:800; abeam) and GAPDH (1:2,000; CST), loading control, antibodies overnight at 4°C, and then anti-rabbit IgG monoclonal antibody conjugated with horseradish peroxidase (Pierce) at 1:2000 dilution for 1 h at room temperature. Protein bands are detected using the West Femto system (Pierce).
  • a decrease in DKK2 mRNA expression in the dermal fibroblast cells is expected after transfection with the DKK2 ASOs compared to DKK2 mRNA levels in dermal fibroblast cells transfected with the non-specific control ASO 48 hours after transfection.
  • mice are divided into four groups: Group 1 - a group treated with non-targeting control siRNA, Group 2 - a group treated with non-targeting control ASO, Group 3 - a group treated with DKK2 siRNA 1, Group 4 - a group treated with DKK2 ASO1. Each group contains eight mice (4 males, 4 females). Each group has an equal portion of the hind limb shaved divided visually into 12 sections. Each section is assessed weekly and given a s.
  • siRNA or ASO Administration of siRNA or ASO is achieved with a topical application of siRNA or ASO resuspended in vehicle at concentration of lOuM to the portion of the mouse skin initially exposed by shaving.
  • Group 1 mice will be treated with non -targeting control siRNA
  • Group 2 mice will be treated with non-targeting control ASO
  • Group 3 mice will be treated with siRNA 1 targeting human DKK2
  • Group 4 mice will be treated with ASO1 targeting human DKK2
  • Group 5 mice will be treated with vehicle.
  • Mice are treated once a week for 7 weeks with the final assessment taken 7 days after final treatment.
  • mice 7 days after the final treatment, the mice are sacrificed by cervical dislocation following an intraperitoneal injection of 0.3 ml Nembutal (5 mg/ml). The shaved portion of skin tissue is collected and stored in RNAlater.
  • mRNA is isolated from tissue placed in RNAlater solution using the PureLink kit according to the manufacturer’s protocol (ThermoFisher Cat. No. 12183020). The reverse transcriptase reaction is performed according to the manufacturer’s protocol. Samples are stored at -80 °C until real-time qPCR is performed in triplicate using TaqMan Gene Expression Assays (Applied Biosystems FAM/DKK2 using a BioRad iCycler). There is an expected decrease in DKK2 mRNA expression in skin tissue from mice dosed with the DKK2 siRNA 1 or ASO1 compared to DKK2 mRNA levels in the skin tissue from mice dosed with the non-specific controls.
  • human subjects with hair loss are treated topically with an siRNA or ASO targeting DKK2, or with a control such as a placebo.
  • a topical formulation comprising the siRNA or ASO is administered to the scalp of the subject.
  • Signs and symptoms of hair loss are observed before, during, and after the treatment. For example, any one or more of the following may be determined: Men's Hair Growth Questionnaire (MHGQ) results, Kingsley Alopecia Profile (KAP) results, total hair counts, vellus hair counts, non-vellus hair counts, hair thickness measurements, hair density measurements, numbers of hair follicles (including total hair follicles, terminal hair follicles, anagen hair follicles, telogen hair follicles, catagen hair follicles, vellus or vellus-like miniaturized hair follicles, and indeterminate hair follicles), and protein and mRNA levels for [3-catenin, a-SMA, collagen I, and collagen III. Additionally, DKK2 protein (circulating and skin) levels and DKK2 mRNA (skin) levels are determined. The topical treatment with either the siRNA or the ASO is expected to improve these measurements.
  • MHGQ Men's Hair Growth Question
  • Oligonucleotides such as siRNAs may be synthesized according to phosphoramidite technology on a solid phase.
  • a K&A oligonucleotide synthesizer may be used. Syntheses may be performed on a solid support made of controlled pore glass (CPG, 500 A or 600 A, obtained from AM Chemicals, Oceanside, CA, USA). All 2'-0Me and 2’-F phosphoramidites may be purchased from Hongene Biotech (Union City, CA, USA). All phosphoramidites may be dissolved in anhydrous acetonitrile (100 mM) and molecular sieves (3 A) may be added.
  • CPG controlled pore glass
  • All phosphoramidites may be dissolved in anhydrous acetonitrile (100 mM) and molecular sieves (3 A) may be added.
  • 5 -Benzylthio- IH-tetrazole (BTT, 250 mM in acetonitrile) or 5-Ethylthio-lH-tetrazole (ETT, 250 mM in acetonitrile) may be used as activator solution. Coupling times may be 9-18 min (e.g. with a GalNAc such as ETL17), 6 min (e.g. with 2'0Me and 2'F).
  • POS 3-phenyl 1,2,4- dithiazoline-5-one
  • POS 3-phenyl 1,2,4- dithiazoline-5-one
  • the dried solid support may be treated with a 1 : 1 volume solution of 40 wt. % methylamine in water and 28% ammonium hydroxide solution (Aldrich) for two hours at 30° C.
  • the solution may be evaporated and the solid residue may be reconstituted in water and purified by anionic exchange HPLC using a TKSgel SuperQ-5PW 13u column.
  • Buffer A may be 20 mM Tris, 5 mM EDTA, pH 9.0 and contained 20% Acetonitrile and buffer B may be the same as buffer A with the addition of 1 M sodium chloride. UV traces at 260 nm may be recorded. Appropriate fractions may be pooled then desalted using Sephadex G-25 medium.
  • Equimolar amounts of sense and antisense strand may be combined to prepare a duplex.
  • the duplex solution may be prepared in 0. 1 xPBS (Phosphate-Buffered Saline, l x, Gibco).
  • the duplex solution may be annealed at 95° C. for 5 min, and cooled to room temperature slowly.
  • Duplex concentration may be determined by measuring the solution absorbance on a UV-Vis spectrometer at 260 nm in 0.1 xPBS. For some experiments, a conversion factor may be calculated from an experimentally determined extinction coefficient.
  • Example 9 GalNAc ligand for hepatocyte targeting of oligonucleotides
  • GalNAc multivalent N-acetylgalactosamine
  • oligonucleotides there are at least two general methods for attachment of multivalent N-acetylgalactosamine (GalNAc) ligands to oligonucleotides: solid or solution-phase conjugations.
  • GalNAc ligands may be attached to solid phase resin for 3’ conjugation or at the 5’ terminus using GalNAc phosphoramidite reagents.
  • GalNAc phosphoramidites may be coupled on solid phase as for other nucleosides in the oligonucleotide sequence at any position in the sequence.
  • the oligonucleotide sequence — including a reactive conjugation site — is formed on the resin.
  • the oligonucleotide is then removed from the resin and GalNAc is conjugated to the reactive site.
  • the carboxy GalNAc derivatives may be coupled to amino-modified oligonucleotides.
  • the peptide coupling conditions are known to the skilled in the art using a carbodiimide coupling agent like DCC (N,N'-Dicyclohexylcarbodiimide), EDC (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide) or EDC.HC1 (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and an additive like HOBt (1- hydroxybenztriazole), HOSu (N -hydroxy succinimide), TBTU (N,N,N',N'-Tetramethyl-O-(benzotriazol-l- yljuronium tetrafluoroborate, HBTU (2-(lH-benzotriazol-l-yl)-l,l,3,3-tetra
  • Amine groups may be incorporated into oligonucleotides using a number of known, commercially available reagents at the 5’ terminus, 3’ terminus or anywhere in between.
  • Non-limiting examples of reagents for oligonucleotide synthesis to incorporate an amino group include:
  • Solution phase conjugations may occur after oligonucleotide synthesis via reactions between non- nucleosidic nucleophilic functional groups that are attached to the oligonucleotide and electrophilic GalNAc reagents.
  • nucleophilic groups include amines and thiols
  • electrophilic reagents include activated esters (e.g. N-hydroxysuccinimide, pentafluorophenyl) and maleimides.
  • Example 10 GalNAc ligands for hepatocyte targeting of oligonucleotides
  • GalNAc multivalent N-acetylgalactosamine
  • oligonucleotides there are at least two general methods for attachment of multivalent N-acetylgalactosamine (GalNAc) ligands to oligonucleotides: solid or solution-phase conjugations.
  • GalNAc ligands may be attached to solid phase resin for 3’ conjugation or at the 5’ terminus using GalNAc phosphoramidite reagents.
  • GalNAc phosphoramidites may be coupled on solid phase as for other nucleosides in the oligonucleotide sequence at any position in the sequence.
  • a non-limiting example of a phosphoramidite reagent for GalNAc conjugation to a 5’ end oligonucleotide is shown in Table 8.
  • the reaction mixture is diluted with DCM (400 mL) and washed with aq.NaHCOs (400 mL * 1) and brine(400 mL * 1), then the mixture is diluted with DCM (2.00 L) and washed with 0.7 M Na2COs (1000 mL * 3) and brine(800 mL * 3), dried over Na2SC>4, filtered and concentrated under reduced pressure to give a residue. The residue is used to next step directly without purification.
  • An example DKK2 siRNA includes a combination of the following modifications:
  • Antisense strand odd-numbered positions are 2'OMe and even-numbered positions are a mixture of 2’ F, 2’OMe and 2’ deoxy.
  • An example DKK2 siRNA includes a combination of the following modifications:
  • Antisense strand odd-numbered positions are 2'OMe and even-numbered positions are a mixture of 2’ F, 2’OMe and 2’ deoxy.
  • Example 13 Topical screening of mouse-human cross-reactive sequence in vivo:
  • RNAlater Total skin RNA was prepared by homogenizing the tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles.
  • RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations.
  • Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions.
  • mice DKK2 mRNA were assessed by RT-qPCR in triplicate on a QuantStudioTM 6 Pro Real-Time PCR System using and the mouse housekeeping gene PPIA (ThermoFisher, assay#Mm02342430_g I ). Data were normalized to the level in animals receiving ETD01043. Results are displayed in Table 9. siRNA sequences are included in Table 6 or 13A.
  • Example 14 Screening DKK2 siRNAs for activity in SH-SY5Y cells in culture
  • the DKK2 siRNAs were individually transfected into SH-SY5Y cells in duplicate wells at 30 nM final concentration using 0.2 pL Lipofectamine RNAiMax (Fisher, catalog# HSS 120398) in 5 pL Opti-MEM (Thermo Fisher, catalog# 31985070) per well. Silencer Select Negative Control #1 (ThermoFisher, catalog# 4390843) was transfected at 30 nM final concentration as a negative control. Positive control siRNAs targeting DKK2 (ThermoFisher) were transfected at 30 nM final concentration.
  • the level of DKK2 mRNA from each well was measured in triplicate by biplex real-time qPCR on a QuantStudio 6 Pro instrument (Applied Biosystems) using TaqMan® Fast Advanced Master Mix (Fisher Scientific catalog# 44-445-58), TaqMan Gene Expression Assay for human DKK2 and TaqMan Gene Expression Assay for human PPIA (ThermoFisher, assay# Hs99999904_ml).
  • the relative DKK2 mRNA levels in each well was calculated using the delta-delta Ct method. All data were normalized to relative DKK2 mRNA levels in untreated SH-SY5Y cells. Results are shown in Table 10. siRNA sequences are included in Table 13A.
  • Example 15 Screening DKK2 siRNAs for activity in A673 cells in culture
  • DKK2 siRNAs cross-reactive for at least human pigs were assayed for DKK2 mRNA knockdown activity in cells in culture.
  • A673 cells were seeded in 96-well tissue culture plates at a cell density of 20,000 cells per well in DMEM media (VWR catalog# 02-0100-0500) supplemented with 10% fetal bovine serum and incubated overnight in a water-jacketed, humidified incubator at 37°C in an atmosphere containing 5% carbon dioxide.
  • the DKK2 siRNAs were individually transfected into A673 cells in duplicate wells at 30 nM final concentration using 0.2 pL Lipofectamine RNAiMax (Fisher, catalog# HSS120398) in 5 pL Opti-MEM (Thermo Fisher, catalog# 31985070) per well. Silencer Select Negative Control #1 (ThermoFisher, catalog# 4390843) was transfected at 30 nM final concentration as a negative control. Positive control siRNAs targeting DKK2 (ThermoFisher) were transfected at 30 nM final concentration.
  • the level of DKK2 mRNA from each well was measured in triplicate by biplex real-time qPCR on a QuantStudio 6 Pro instrument (Applied Biosystems) using TaqMan® Fast Advanced Master Mix (Fisher Scientific catalog# 44-445-58), TaqMan Gene Expression Assay for human DKK2 and TaqMan Gene Expression Assay for human PPIA (ThermoFisher, assay# Hs99999904_ml).
  • the relative DKK2 mRNA levels in each well was calculated using the delta-delta Ct method. All data were normalized to relative DKK2 mRNA levels in untreated A673 cells. Results are shown in Table 11. siRNA sequences are included in
  • Example 16 Screening siRNAs targeting human DKK2 mRNA in mice transfected with AAV8- TBG-h-DKK2
  • siRNAs targeting human DKK2 mRNA and cross-reactive with at least pig DKK2 mRNA were tested for activity in mice following transfection with an adeno-associated viral vector.
  • Six- to eight-week-old female mice (C57B1/6) were injected with 5 pL of a recombinant adeno-associated virus 8 (AAV8) vector (8.9 x 10E12 genome copies/mL) by the retroorbital route on Day -14.
  • AAV8 vector recombinant adeno-associated virus 8
  • the recombinant AAV8 contained the open reading frame and the majority of the 3’UTR of the human DKK2 sequence (NM_014421.3) under the control of the human thyroxine binding globulin promoter in an AAV2 backbone packaged in AAV8 capsid (AAV8-TBG-h-DKK2 ).
  • RNAlater ThermoFisher Catalog# AM7020
  • Total liver RNA was prepared by homogenizing the liver tissue in homogenization buffer (Maxwell RSC simplyRNA Tissue Kit) using a Percellys 24 tissue homogenizer (Bertin Instruments) set at 5000 rpm for two 10 second cycles.
  • Total RNA from the lysate was purified on a Maxwell RSC 48 platform (Promega Corporation) according to the manufacturer’s recommendations.
  • Preparation of cDNA was performed using Quanta qScript cDNA SuperMix (VWR, Catalog# 95048-500) according to the manufacturer’s instructions.
  • liver DKK2 mRNA The relative levels of liver DKK2 mRNA were assessed by RT-qPCR in triplicate on a QuantStudioTM 6 Pro Real-Time PCR System using TaqMan assays for human DKK2 and the mouse housekeeping gene PPIA (ThermoFisher, assay# Mm02342430_g I ) and PerfeCTa® qPCR FastMix®, Low ROXTM (VWR, Catalog# 101419-222). Data were normalized to the mean DKK2 mRNA level in animals receiving PBS. Results are shown in Table 12. siRNA sequences are included in Table 13A.
  • Example 18 siRNA treatment of donor hair follicles
  • HFs Human amputated hair follicles
  • FUE follicular unit extract
  • Oxcipital HFs from FUE
  • HFs were cultured at 37° C with 5% CO2 in a minimal media of William's E media (Gibco, Life Technologies) supplemented with 2 mM of L-Glutamine (Gibco), 10 ng/ml hydrocortisone (Sigma- Aldrich), 10 pg/ml insulin (Sigma- Aldrich), and 1% penicillin and streptomycin mix (Gibco) to make William's complete media (WCM).
  • WCM William's complete media
  • follicles were assigned per group. 24 h after isolation, WCM was replaced, and HFs were treated with siRNA’s until Day 3 of culture. On day 3, the culture medium was replaced. For groups 5 and 6 on day 3, the culture medium was replaced and there was reapplication of media and test articles. On days 4 (Groups 1-4) and 5 (Groups 5 and 6) follicles were harvested by placement in RNA extraction buffer and stored at -80 °C.
  • RNA isolation was performed using the Arcturus PicoPure RNA isolation kit (KIT0204) from Applied Biosystems following the manufacturer’s protocol. cDNA synthesis
  • cDNA synthesis was performed using a Tetro cDNA Synthesis Kit (BIO-65043) from Meridian Bioscience following the manufacturer’s protocol.
  • TaqMan qPCR was performed with TaqMan Fast Advanced Master Mix (4444556;

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Abstract

La divulgation concerne des compositions comprenant un oligonucléotide ciblant Dickkopf2 (DKK2). L'oligonucléotide peut comprendre un petit ARN interférent (ARNsi) ou un oligonucléotide antisens (ASO). La divulgation concerne également des procédés de réduction de l'expression de DKK2 par fourniture d'un oligonucléotide ciblant DKK2 à un sujet en ayant besoin. Certains modes de réalisation comprennent des méthodes de traitement de la perte de cheveux par fourniture de l'oligonucléotide.
PCT/US2023/084879 2022-12-20 2023-12-19 Traitement de maladies et de troubles liés au dkk2 Ceased WO2024137663A2 (fr)

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KR101508052B1 (ko) * 2013-05-20 2015-04-08 울산대학교 산학협력단 생물학적 활성을 가진 인간 Ｄｋｋ２ 재조합 단백질의 수용성 발현 및 정제방법
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