WO2012149406A2 - Estimation et traitement d'êtres humains atteints d'un syndrome du qt long - Google Patents

Estimation et traitement d'êtres humains atteints d'un syndrome du qt long Download PDF

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WO2012149406A2
WO2012149406A2 PCT/US2012/035575 US2012035575W WO2012149406A2 WO 2012149406 A2 WO2012149406 A2 WO 2012149406A2 US 2012035575 W US2012035575 W US 2012035575W WO 2012149406 A2 WO2012149406 A2 WO 2012149406A2
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kcnql
mutation
utr
exon
kcnq
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WO2012149406A3 (fr
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Michael J. Ackerman
John R. GIUDICESSI
Yigal Martin Pinto
Ahmad Shoaib AMIN
Anke Johanna Marina Tijsen
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Academisch Medisch Centrum
Mayo Foundation for Medical Education and Research
Mayo Clinic in Florida
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Mayo Foundation for Medical Education and Research
Mayo Clinic in Florida
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Definitions

  • This document relates to methods and materials involved in assessing and treating humans with long QT syndrome (LQTS) or with a potential mutation in a KCNQ 1 nucleic acid that encodes a K v 7.1 potassium channel subunit.
  • LQTS long QT syndrome
  • this document provides methods and materials for determining if a human containing a mutation in a KCNQ1 nucleic acid that encodes a K v 7.1 potassium channel subunit on one allele also contains, on the same allele (a cis relationship) or on the other allele (a trans relationship), a genetic variation (e.g., a SNP) in a 3 ' UTR of KCNQ 1 nucleic acid that creates a new microRNA (e.g., miR-378) binding site.
  • LQTS long QT syndrome
  • LQTS is a common heritable cardiac channelopathy that can cause premature sudden death due to life-threatening arrhythmias in relation to prolongation of the heart rate-corrected QT interval (QTc).
  • the most prevalent form, LQT1 is caused by loss-of- function mutations in the KCNQ 1 -encoded K v 7.1 potassium channel (IKs).
  • LQT1 is characterized by incomplete penetrance and variable expressivity whereby family members carrying identical mutations have profound differences in their QTc and clinical course. The cause for this heterogeneity remains largely elusive.
  • K v 7.1 is a tetrameric channel derived from the post-translational assembly of four KCNQ 1 -encoded subunits.
  • patients heterozygous for an LQT1 -causative mutation combine the translated products from normal and LQT1 -mutation-containing alleles to form tetrameric channels. If both alleles are similarly expressed, one would predict that 1/16th of the K v 7.1 channels stem solely from the normal allele, and 1/16th of the K v 7.1 channels stem solely from the mutated allele. The remaining channels would be hybrids of the mutated and healthy alleles.
  • This document relates to methods and materials involved in assessing and treating humans with LQTS or with a potential mutation in a KCNQ 1 nucleic acid that encodes a K v 7.1 potassium channel subunit. For example, this document provides methods and materials for determining if a human containing a mutation in a KCNQ 1 nucleic acid that encodes a K v 7.1 potassium channel subunit on one allele also contains, on the same allele (a cis relationship) or on the other allele (a trans relationship), a genetic variation (e.g., a SNP) in the 3' UTR of KCNQl nucleic acid that creates a new microRNA (e.g., a miR- 378) binding site.
  • a genetic variation e.g., a SNP
  • KCNQ 1 mutation refers to a genetic variation (e.g., a substitution, deletion, or insertion of a nucleotide or nucleotides) that is present within a KCNQl nucleic acid that encodes a K v 7.1 potassium channel subunit, causes an alteration in the amino acid sequence of a K v 7.1 potassium channel subunit, and is associated with LQTS.
  • KCNQl UTR variation refers to a genetic variation (e.g., a substitution, deletion, or insertion of a nucleotide or nucleotides) that is present within the 3' untranslated region of a KCNQl nucleic acid and creates a microRNA binding site (e.g., a miR-378 binding site).
  • a KCNQ l mutation that is present on the same allele that contains a KCNQ 1 UTR variation can experience less severe symptoms of LQTS if the other allele lacks KCNQl mutations (e.g., is a wild-type KCNQl nucleic acid).
  • KCNQl nucleic acid lacking KCNQl mutations e.g., a wild-type KCNQl nucleic acid that encodes a K v 7.1 potassium channel subunit
  • KCNQl UTR variation Humans with KCNQl nucleic acid lacking KCNQl mutations (e.g., a wild-type KCNQl nucleic acid that encodes a K v 7.1 potassium channel subunit) that is present on the same allele that contains a KCNQl UTR variation can experience more severe symptoms of LQTS if the other allele contains a KCNQl mutation.
  • the cis or trans relationship of KCNQ 1 mutations and KCNQ 1 UTR variations can be used to assess the severity of LQTS.
  • Having the ability to determine the cis or trans relationship of KCNQ 1 mutations and KCNQl UTR variations within a human heterozygous for a KCNQl mutation and heterozygous for a KCNQ 1 UTR variation can allow clinicians and patients to determine appropriate levels of LQTS disease monitoring and care. For example, a patient identified as having increased LQTS severity as described herein can be selected for treatment with agents designed to inhibit microRNA activity.
  • this document features a method for assessing the cis or trans nature of a human heterozygous for a KCNQ 1 mutation and heterozygous for a KCNQ 1 UTR variation.
  • the method comprises, or consists essentially of, (a) determining if an allele of the human comprises (i) the KCNQ 1 mutation and the KCNQ 1 UTR variation, (ii) a lack of the KCNQl mutation and a lack of the KCNQl UTR variation, (iii) the KCNQl mutation and a lack of the KCNQl UTR variation, or (iv) the KCNQl UTR mutation and a lack of the KCNQl variation, (b) classifying the human as having the KCNQl mutation and the KCNQl UTR variation in cis if the allele comprises (i) or (ii), and (c) classifying the human as having the KCNQl mutation and the KCNQl UTR variation in trans if the allele comprises (iii) or (
  • the human can be an LQT1 patient.
  • the allele can comprise the KCNQ 1 mutation and the KCNQ 1 UTR variation, and the human can be classified as having the KCNQ l mutation and the KCNQl UTR variation in cis.
  • the allele can comprise a lack of the KCNQl mutation and a lack of the KCNQl UTR variation, and the human can be classified as having the KCNQl mutation and the KCNQl UTR variation in cis.
  • the allele can comprise the KCNQl mutation and a lack of the KCNQl UTR variation, and the human can be classified as having the KCNQl mutation and the KCNQl UTR variation in trans.
  • the allele can comprise the KCNQl UTR variation and a lack of the KCNQ 1 mutation, and the human can be classified as having the KCNQl mutation and the KCNQl UTR variation in trans.
  • the KCNQl mutation can be L266P, R518X, G168R, or R594Q.
  • the KCNQl UTR variation can be an rs2519184 SNP or rs8234 SNP.
  • this document features a method for treating a mammal having a KCNQl UTR variation that is part of a microRNA binding site, wherein the binding of a microRNA to the binding site reduces expression of a K v 7.1 potassium channel subunit lacking a mutation associated with LQTS.
  • the method comprises, or consists essentially of, administering a miRNA inhibitor to the mammal under conditions wherein the miRNA inhibitor reduces the ability of the microRNA to reduce expression of the subunit.
  • this document features a method for treating a human comprising, or consisting essentially of, administering an agent that inhibits or mimics the activity of a microR A that targets a KCNQ1 UTR variation to increase the ratio of expression of a non-mutated KCNQ 1 allele to mutated KCNQ 1 allele.
  • this document features a method for treating a human having long QT syndrome.
  • the method comprises, or consists essentially of, (a) determining if an allele of the human comprises (i) the KCNQ1 mutation and the KCNQ1 UTR variation, (ii) a lack of the KCNQ 1 mutation and a lack of the KCNQ 1 UTR variation, (iii) the KCNQ1 mutation and a lack of the KCNQ1 UTR variation, or (iv) the KCNQ1 UTR mutation and a lack of the KCNQ 1 variation, and (b) administering a beta blocker to the human, implanting an implantable cardioverter-defibrillator into the human, or performing a sympathetic denervation procedure on the human.
  • the method can comprise administering the beta blocker to the human.
  • the beta blocker can be metoprolol, carvedilol, bisoprolol, nebivolol, or propanolol.
  • the method can comprise implanting the implantable cardioverter-defibrillator into the human.
  • the method can comprise performing the sympathetic denervation procedure on the human.
  • FIG. 1 Genetic variation in the 3 'untranslated region oiKCNQl.
  • SNPs Single nucleotide polymorphisms
  • AMC Academic Medical Center Amsterdam
  • MC Mayo Clinic
  • Position of the nucleotide change is starting from the ATG start codon ( CBI build 36, hgl 8).
  • B Predicted binding between miR-378 and the 3'UTR oiKCNQl containing the minor A variant of SNP rs2519184 and the minor G variant of SNP rs8234. 'rs' numbers denote SNP identities from public database. Longer lightly shaded boxes represent exons, and darker shaded bars represent SNPs.
  • C Relative expression of miR-378 in human donor hearts in comparison with other abundantly expressed cardiac miRNAs (miR-133b, miR-21, and miR-208b). Data are presented as mean ⁇ standard error.
  • FIG. 1 The functional effects of genetic variation in the 3 'untranslated region (3 'UTR) oiKCNQl.
  • the dashed line represents mean QTc of all individuals within the study population regardless of the specific LQT1 -causative mutation and the 3 'UTR SNP status. Numbers below genotypes denote group sizes. Data are presented as mean ⁇ standard error. N, normal KCNQ1 allele; M, mutant KCNQ1 allele. Darker shaded box represents a major SNP variant, while the lighter shaded box represents a minor SNP variant.
  • (3 'UTR) oiKCNQl (A) Luciferase assays in neonatal rat cardiomyocytes transfected with two independent reporter plasmids containing either the major or the minor haplotype of SNPs in the 3'UTR oiKCNQl. Observed differences in luciferase activity between the major and minor haplotype may indicate that translation is inhibited by enhanced miRNA binding to the 3 'UTR.
  • Figure 7 is a diagram of a mechanism where SNPs in the 3 'UTR oiKCNQl modulate the assembly of the K v 7.1 potassium channel in type 1 long QT syndrome.
  • Individuals with type long QT syndrome (LQT1) are heterozygous for the disease- causing mutation in KCNQ1.
  • Four KCNQ1 -encoded subunits co-assemble post- trans lationally to form one tetrameric channel. Since the 3 'UTR plays an important role in messenger-RNA translation, SNPs in this region may influence repolarization by altering the balance and composition of the K v 7.1 tetramers derived from translation of the normal allele and the mutant allele.
  • the balance between normal and mutant subunits within each channel is equal. However, if the minor genotypes of the SNPs suppress messenger-RNA translation (by creating target sites for microRNAs as predicted for SNPs rs2519184 and rs8234; see Figure 2), then the balance between normal and mutant subunits within each channel depends on whether the 'suppressive SNPs' reside on the normal allele or the mutant allele. If the 'suppressive SNPs' reside on the normal allele, the number of normal subunits in the channels would decrease.
  • Figure 8 is a graph showing inhibition by a microRNA inhibitor (e.g., an anti- miR) designed to inhibit miR-378, at two different dosages.
  • a microRNA inhibitor e.g., an anti- miR
  • Figure 9 is a graph showing that the inhibition shown in Figure 8 prevents the suppressive effects of UTR variants.
  • the two bars on the right (antimiR54) represent a control demonstrating that the effect (i.e., lack of suppression) only occurs after using the specific antimir against 378.
  • Figure 10 is a graph of luciferase assay results of cardiac myocytes with knockdown of miR-378.
  • Figure 1 1 is a graph demonstrating an allelic imbalance on mRNA levels.
  • This document relates to methods and materials involved in assessing and treating humans with LQTS or with a potential mutation in a KCNQ1 nucleic acid that encodes a K v 7.1 potassium channel subunit.
  • this document provides methods and materials for assessing LQTS severity by determining if a human containing a KCNQ1 mutation on one allele also contains, on the same allele (a cis relationship) or on the other allele (a trans relationship), a KCNQ 1 UTR variation.
  • Figure 7 provides several examples of how the cis and trans relationship of KCNQ 1 mutations and KCNQ 1 UTR variations can influence LQTS severity.
  • KCNQ 1 mutations include, without limitation, those listed in Table A or described elsewhere (Kapplinger et ah, Heart Rhythm., 6(9): 1297-303 (2009)).
  • KCNQ 1 UTR variations include, without limitation, the rs2519184 SNP, rs8234 SNP, and rs 10798 SNP.
  • a KCNQ1 UTR variation can be any genetic variant present within the 3' untranslated region of a KCNQ1 nucleic acid that creates a microRNA binding site for a microRNA that is capable of reducing expression of mRNA to which it binds. Examples of such microRNA include, without limitation, miR-378, miR-328, and miR-220. Table A. KCNQ1 LQT1 -associated mutations.
  • KCNQl niRNA e.g., KCNQl cDNA generated by RT-PCR
  • KCNQl cDNA generated by RT-PCR can be sequenced to determine the cis or trans relationship of KCNQl mutations, KCNQl UTR variations, or wild-type sequences.
  • Any appropriate biological sample such as a blood lymphocytes, skin biopsy, or myocardial biopsy can be used as a source for KCNQ l mRNA.
  • a person identified as having a more severe form of LQTS can be at an increased risk for sudden death.
  • the person can be treated prophylactically with, for example, beta blocker therapy, ICD implantation, and/or sympathetic denervation.
  • a human determined to have a cis or trans relationship of KCNQ l mutations, KCNQl UTR mutations, or wild-type sequences indicative of more severe LQTS can be treated with one or more microRNA inhibitors (e.g., anti-miRs) designed to reduce the ability of a microRNA (e.g., miR-378) to inhibit the expression of a wild-type
  • a microRNA inhibitor can be designed to inhibit or reduce the activity of a particular microRNA (e.g., a miR-378).
  • a particular microRNA e.g., a miR-378
  • anti-miRs designed to inhibit or reduce the activity of miR-378 include, without limitation, cholesterol-based antagomiR agents and locked nucleic acid-based anti-miRs.
  • an anti-miR can have various backbone and/or sugar modifications such as 2'-0-methyl (2'-OMe), 2'-fluoro (2'
  • Example 1 Functional variants in KCNQl 's 3 'UTR modify disease severity in patients with Type 1 LOTS through miR A-dependent mechanisms Genetic Analysis
  • LQT1 -associated KCNQ1 mutations were identified previously using standard protocols (Bhuiyan et ah, Prog. Biophys. Mol. Biol, 98(2-3):319-327 (2008)). For this analysis, the final exon and the 3'UTR of KCNQ1 were sequenced.
  • the region to be analyzed included 3 amplicons and was amplified with polymerase chain reaction using oligonucleotide pairs: 5'-GGCACCTTCCCTTCTCTGG-3' (SEQ ID NO: 1) with 5'-ACC- ACCATGCCAGTGATGTC-3 ' (SEQ ID NO:2); 5'-CACAGCCTGCACTTGGG-3' (SEQ ID NO:3) with 5'-CAGGGCTCCTCTCCAGC-3' (SEQ ID NO:4); and 5'-CAGTCTCA- CCATTTCCCCAG-3' (SEQ ID NO: 5) with 5 '-GCCCAGAACAGGAGCGAC -3' (SEQ ID NO: 6).
  • the KCNQ1 3 'UTR mRNA sequence was retrieved from the Ensemble Genome Browser (NCBI build 36, hgl 8), and SNPs identified experimentally in a KCNQ1 mutation carrier were mapped on the KCNQ 1 mRNA sequence. For each SNP, two sequences centered on the major or minor allele with 30 nucleotides flanking each side were assembled. The search by sequence option of www.mirbase.org was used to identify potential miRNA binding sites within these sequences using an E-value cut-off of 1000. This option used the miRanda algorithm to predict targeting of all miRNAs in the database to the input sequence. Comparison of the major and minor allele for miRNA target site predictions for each SNP was used to determine potential miRNA target sites that were created by the presence of the minor allele. miRNA Expression in Human Hearts
  • a luciferase reporter plasmid was constructed by amplifying the 3'UTR of KCNQ1 from a patient heterozygous for the SNPs rs2519184, rs8234, and rsl0798 with the following primers: 5 ' -ACTGACTAGTCATGGACC- ATGCTGTCTG-3 ' (SEQ ID NO: 7) and 5 ' -ACTGGAGCTCCAGCCTGTGATTCTC- CACG-3' (SEQ ID NO: 8).
  • This 878 base pair fragment was cloned into the pMIR- REPORTTM Luciferase vector (Ambion) downstream of the luciferase coding region, creating luciferase-KCNQl-3'UTR-G-A-A (major haplotype) and luciferase-KCNQ 1 - 3'UTR-A-G-G (minor haplotype).
  • the miR-378 overexpression vector, pCDHl-miR- 378 was constructed by PCR-amplification of miR-378 precursor DNA form human genomic DNA using the following primers: 5 '-ACTGGAATTCAGAAAGAGGCTG- CGAGGAG-3 ' (SEQ ID NO: 9) and 5 ' -ACTGGGATCCGGAACAACCAGAAC- ATCTCAC-3' (SEQ ID NO: 10). This 306 base pair fragment was cloned into the pCDHl-MCSl-EFl-Puro vector (System Biosciences) under the control of a CMV promotor.
  • the negative control miRNA overexpression vector, PCDHl-control-miRNA was based on the pcDNATM6.2-GW/miR-neg control plasmid (Invitrogen), which contained an insert that can form a hairpin structure that is processed into mature miRNA but is predicted not to target any known vertebrate gene. All generated constructs were verified by sequencing.
  • Neonatal rat cardiac myocytes immortalized with a temperature-sensitive SV40 T antigen (H10 cells) as described elsewhere (Jahn et ah, J. Cell Set, 109(Pt 2):397-407 (1996)), were cultured in Dulbecco's Modified Eagles Medium supplemented with 10% fetal calf serum (Gibco-BRL) and glutamine at 33 °C.
  • H10 cells Dulbecco's Modified Eagles Medium supplemented with 10% fetal calf serum (Gibco-BRL) and glutamine at 33 °C.
  • Gibco-BRL fetal calf serum
  • One day prior to transfection cells were seeded in a 24-well plate at a density of 1.4xl0 6 cells per plate. Cardiomyocytes from 1-2-day-old Lewis neonatal rats were isolated and cultured as described elsewhere (Leenders et al, J. Biol. Chem., 285(35):274
  • H10 cells were transiently transfected per well with 5 ng Renilla luciferase plasmid, phRL vector (Promega), and 5 ng or 10 ng of either luciferase-KCNQ 1 -3 'UTR- G-A-A, luciferase-KCNQ 1 -3 'UTR-A-A-A, luciferase-KCNQ 1 -3 'UTR-G-G-A, or luciferase-KCNQ 1 -3 'UTR-A-G-G using GeneJammer (Agilent Technologies).
  • Neonatal rat cardiomyocytes were transfected with 50 ng Renilla luciferase plasmid, phRL vector, and 100 ng of either luciferase-KCNQ 1 -3 'UTR-G-A-A, luciferase-KCNQ 1-3 'UTR-A-A- A, luciferase-KCNQ 1-3 'UTR-G-G-A, or luciferase-KCNQ 1-3 'UTR-A-G-G using lipofectamine 2000 reagent (Invitrogen).
  • COS-7 cells were transiently transfected per well with 5 ng Renilla luciferase plasmid, phRL vector, 100 ng of either luciferase- KCNQ 1-3 'UTR-G-A-A or luciferase-KCNQ 1-3 'UTR-A-G-G, 100 ng of miR-378 or negative control miRNA overexpression construct, and empty pCDHl as filler using polyethylenimine (PEI).
  • PEI polyethylenimine
  • SNPs are present in the 3 'UTR of KCNQl
  • P448fsX13 frameshift mutation whereby P448 represents the last normally encoded amino acid followed by a frameshift (fs) in the coding sequence generating 13 miscoded amino acids leading up to a premature stop codon (X); 387- ?_1393+?del, mutation leading to the deletion of exon 2 (starting from nucleotide 387) to exon 10 (ending at nucleotide 1393), but whereby the exact nucleotide change is not yet determined (?); N-terminus, amino-terminus; S, transmembrane segment; C-terminus, carboxyl-terminus; SAD, subunit assembly domain; Cr., center in which the mutation was found (A: Academic Medical Center Amsterdam, M: Mayo Clinic); #, number of individuals within each study population affected with the mutation; Ref, reference linking the mutation with long QT syndrome; *, biophysical/loss-of-function properties of the mutation are described in the reference.
  • the SNP ID denotes single nucleotide polymorphism identity from public
  • N (%) denotes number of patients (percentage of total). Minor variants of SNPs rs2519184 and rs8234 in KCNQ1 's 3 'UTR create binding sitesor miRNA-378
  • mirbase.org World Wide Web at "mirbase.org" predicted that the minor (A) variant of SNP rs2519184 and the minor (G) variant of SNP rs8234 create binding sites for a number of miRNAs including two binding sites for miRNA-378 (miR- 378; Figure IB). miR-378 was found to be abundantly expressed in the human left
  • the plasmid containing the minor 3 'UTR SNP haplotype (A-G-G), which harbors two created binding sites for miR-378, had significantly lower luciferase activity in both cardiac cell types. Moreover, introduction of either minor allele of SNP rs2519184 or rs8234 into the major allele was sufficient to decrease luciferase activity, indicating that those two nucleotides control translation oiKCNQl ( Figure 2B for H10 cells, and Figure 6B for neonatal rat cardiomyocytes).
  • miR- 378 was overexpressed in COS-7 cells transfected with a reporter containing either the major 3'UTR SNP haplotype (G-A-A) or the minor SNP haplotype (A-G-G).
  • the quantitative PCR was performed using LightCycler 480 sybr green I master (Roche).
  • Allele-specific reverse primers (5'-ACCACAAAT- TATTGATTTCTATGCGAT-3 ' (SEQ ID NO: 1 1) for amplifying the major A allele or 5 ' -ACCACAAATTATTGATTTCTATGCGAC-3 ' (SEQ ID NO: 12) for amplifying the minor G allele
  • a general forward primer (5'-AGCCAGCCAAACACACAG-3 ' (SEQ ID NO: 13)
  • Quantitative PCR reactions were performed on a HghtCycler480 system II (Roche) using the following program: 5 minutes pre-incubation at 95°C and 40 cycles of 10 seconds of denaturation at 95°C, 20 seconds of annealing at 60°C, and 20 seconds of elongation at 72°C. Data were analyzed using LinRegPCR quantitative PCR data analysis software.
  • the starting concentrations of transcripts estimated by this software were corrected for the estimated starting concentrations of the housekeeping gene GAPDH (5'-ACCCACTCCTCCACCTTTGAC-3' (SEQ ID NO: 14) and 5'- ACCCTGTTGCTGTAGCCAAATT-3 ' (SEQ ID NO: 15)) and for the estimated starting concentration of a total KCNQ1 amplicon closely resembling the allele-specific amplicons (5 ' -GAAGTGACGGTTCCTACAC-3 ' (SEQ ID NO: 16) and 5'- AGCTTGCACAATTAATAATCAAAATC-3 ' (SEQ ID NO: 17)).
  • Allele-specific haplotype analysis of SNPs rs2519184 and rs8234 revealed that location of the suppressive minor alleles of the SNPs (A-G) in trans to the mutation was associated with 49 ⁇ 16 milliseconds longer QTc in the AMC cohort and a 60 ⁇ 14 milliseconds longer QTc in the MC cohort ( Figure 3C and Figure 3F).
  • Subjects where the A-G-G haplotype was in cis to the mutation were only present in the MC cohort. Indeed, in these subjects, the opposite effect on QTc was suggested, as the QTc was now attenuated by 12 ⁇ 9 milliseconds.
  • Various allele-specific haplotype combinations of KCNQ1 and the minor alleles of the three SNPs displayed an intermediate effect on QTc.
  • SNPs rs2519184 and rs8234 modify QTc in an allele-specific fashion in singles LQT1 families
  • SNPs can predict disease severity of a given KCNQ1 mutation in one single family.
  • allele-specific haplotype of SNPs rs2519184 and rs8234 with regard to QTc in the largest families in the AMC (14 affected) and MC (20 affected) cohorts were analyzed.
  • the KCNQ1-R243C carriers, where the minor suppressive alleles of one or two SNPs was found in trans to the mutation exhibited markedly longer QTc (533 ⁇ 24 and 489 ⁇ 25 milliseconds, respectively) than their KCNQl-R243C-positive relatives who did not carry these SNPs (441 ⁇ 7 milliseconds; Figure 4A).
  • the effect of SNPs rs2519184, rs8234, and rs 10798 on QTc in the general population To study the effect of SNPs rs2519184, rs8234, and rs l0798 in the general population, the following was performed using the KORA Study to evaluate whether the SNPs rs2519184, rs8234, or rs 10798 are correlated with QTc duration in the general population.
  • the KORA Study is a series of independent population-based
  • the effect on the QTc duration refers to the dosage (i.e., expected number of copies) of the major allele.
  • the QTc duration is calculated by the Bazett's formula, and is corrected for age and gender. The results provided herein demonstrate that SNPs in the 3 'UTR of KCNQ1
  • SNPs rs2519184 and rs8234 create binding sites for miR-378, a miRNA appreciably expressed in the human heart. Creation of these binding sites can allow miR-378 to suppress translation of the SNP-containing KCNQ1 allele. These results are clinically relevant when these suppressive 3 'UTR SNPs occur in carriers of an LQT1 -causing mutation.
  • the allelic location of these suppressive SNPs can alter the balance between normal and mutated K v 7.1 channel subunits.
  • the SNPs are in cis to the mutation, the diseased allele can be suppressed and the expressed LQT1 phenotype can be less severe.
  • the suppressive SNPs reside on the normal non-mutated allele (in trans), then translation of the normal allele can be suppressed and the LQT1 phenotype can be more severe.
  • LQT1 genetic testing can include an analysis and cis/trans phase determination of the suppressive 3 'UTR SNPs in KCNQ 1. More generally, the results provided herein demonstrate that genetic variation in the 3 'UTR may be an important source for clinical variability by altering the balance of translation between the two alleles. It is noteworthy that the size of the effects on QTc duration and symptoms that is described go well beyond what has been shown for modifiers described elsewhere (Pfeufer et ah, Circ. Res., 96(6):693-701 (2005); and Crotti et ah, Circulation,

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Abstract

Cette invention concerne des matériels et méthodes impliqués dans l'estimation et le traitement d'êtres humains atteints de LQTS ou ayant une mutation potentielle dans un acide nucléique KCNQ1 qui code pour une sous-unité du canal potassique Kv7.1. Par exemple, l'invention concerne des matériels et méthodes servant à déterminer si un être humain ayant une mutation dans un acide nucléique KCNQ1 qui code pour une sous-unité du canal potassique Kv7.1 sur un allèle possède également, sur le même allèle (une relation cis) ou sur l'autre allèle (une relation trans), une variation génétique (par exemple un SNP) dans un 3' UTR d'un acide nucléique KCNQ1 qui crée un site de liaison à miR-378.
PCT/US2012/035575 2011-04-29 2012-04-27 Estimation et traitement d'êtres humains atteints d'un syndrome du qt long Ceased WO2012149406A2 (fr)

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WO2012149406A3 WO2012149406A3 (fr) 2013-01-17

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105385688A (zh) * 2015-11-05 2016-03-09 宁波市医疗中心李惠利医院 一种与长QT综合征相关的miRNA及其应用
WO2022147249A1 (fr) * 2020-12-30 2022-07-07 Mayo Foundation For Medical Education And Research Thérapie génique de suppression-remplacement
WO2023191630A1 (fr) * 2022-03-30 2023-10-05 Academisch Medisch Centrum Acides nucléiques antisens destinés à être utilisés dans le traitement de porteurs de mutation de kcnq1

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7179597B2 (en) * 2000-04-13 2007-02-20 Georgetown University Genetic diagnosis for QT prolongation related adverse drug reactions
US7537928B2 (en) * 2003-08-22 2009-05-26 Masonic Medical Research Laboratory Mutations in ion channel proteins associated with sudden cardiac death
US20050142591A1 (en) * 2003-10-29 2005-06-30 Ackerman Michael J. Method of genetic testing in heritable arrhythmia syndrome patients
ITMI20051047A1 (it) * 2005-06-07 2006-12-08 Irccs Fond Salvatore Maugeri C Mutazioni associate alla sindrome del qt lungo e loro uso diagnostico

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105385688A (zh) * 2015-11-05 2016-03-09 宁波市医疗中心李惠利医院 一种与长QT综合征相关的miRNA及其应用
WO2022147249A1 (fr) * 2020-12-30 2022-07-07 Mayo Foundation For Medical Education And Research Thérapie génique de suppression-remplacement
WO2023191630A1 (fr) * 2022-03-30 2023-10-05 Academisch Medisch Centrum Acides nucléiques antisens destinés à être utilisés dans le traitement de porteurs de mutation de kcnq1

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