ES2315040B2 - PROCEDURE TO DETERMINE THE EFFECTIVENESS OF THE TREATMENT AND THE PROGRESSION DEGREE OF CHRONIC MYELOID LEUKEMIA BY USING SPI-1 / PI.1. - Google Patents
PROCEDURE TO DETERMINE THE EFFECTIVENESS OF THE TREATMENT AND THE PROGRESSION DEGREE OF CHRONIC MYELOID LEUKEMIA BY USING SPI-1 / PI.1. Download PDFInfo
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Abstract
Procedimiento para determinar la eficacia del tratamiento y el grado de progresión de la leucemia mieloide crónica mediante el uso de SPI-1/PU.1 que consiste en la determinación de mRNA o de proteína del gen SPI-1/PU.1, en muestras de células de sangre o médula ósea de pacientes de LMC y su comparación con muestras de sujetos sanos o del mismo paciente tras el tratamiento antileucémico. Niveles de mRNA o proteína de SPI-1/PU.1 altos o comparables a los de sujetos sanos son indicadores de respuesta al tratamiento. La presencia de SPI-1/PU.1 es indicador de respuesta al tratamiento y recuperación de hematopoyesis normal. Por el contrario, una expresión reducida es indicador de persistencia de la leucemia y mal pronóstico.Procedure to determine the efficacy of the treatment and the degree of progression of chronic myeloid leukemia through the use of SPI-1 / PU.1 consisting of the determination of mRNA or protein of the SPI-1 / PU.1 gene, in samples of blood or bone marrow cells of CML patients and their comparison with samples from healthy subjects or the same patient after antileukemic treatment. SPI-1 / PU.1 mRNA or protein levels high or comparable to those of healthy subjects are indicators of treatment response. The presence of SPI-1 / PU.1 is an indicator of response to treatment and recovery of normal hematopoiesis. On the contrary, a reduced expression is an indicator of leukemia persistence and poor prognosis.
Description
Procedimiento para determinar la eficacia del tratamiento y el grado de progresión de la leucemia mieloide crónica mediante el uso de SPI-1/PU.1.Procedure to determine the efficacy of the treatment and the degree of progression of chronic myeloid leukemia by using SPI-1 / PU.1 .
Sector de la invención: Biomedicina. Marcadores de pronóstico de la enfermedad y como guía de los profesionales médicos para seleccionar o evaluar los tratamientos de leucemia mieloide crónica.Sector of the invention: Biomedicine. Markers of prognosis of the disease and as a guide for professionals doctors to select or evaluate leukemia treatments chronic myeloid
El factor de transcripción SPI-1/PU.1 es una proteína de la familia ETS, que juega un papel clave en la hematopoyesis. SPI-1/PU.1 regula la expresión de genes importantes para la diferenciación mieloide y linfoide tales como los receptores de los factores de crecimiento de macrófagos, de granulocitos, la integrina CD11b y mieloperoxidasa, así como su propia expresión. También regula la expresión de genes de inmunoglobulinas (Friedman, 2002).The transcription factor SPI-1 / PU.1 is an ETS family protein, which plays a key role in hematopoiesis. SPI-1 / PU.1 regulates the expression of genes important for myeloid and lymphoid differentiation such as macrophage, granulocyte growth factor receptors, CD11b integrin and myeloperoxidase, as well as their own expression. It also regulates the expression of immunoglobulin genes (Friedman, 2002).
Diversos estudios indican un papel fundamental de Spi-1 en el compromiso de las células pluripotentes hacia distintos linajes hematopoyéticos. Los datos con ratones deficientes en SPI-1/PU.1 indican que esta proteína se requiere para el desarrollo tanto de la serie mieloide (granulocitos y monocitos) como linfoide (Lloberas et al., 1999). En progenitores eritroides la expresión normal de Spi-1 es baja y su sobreexpresión desregulada da lugar a eritroleucemia.Several studies indicate a fundamental role of Spi-1 in the involvement of pluripotent cells towards different hematopoietic lineages. Data with mice deficient in SPI-1 / PU.1 indicate that this protein is required for the development of both the myeloid series (granulocytes and monocytes) and lymphoid (Lloberas et al ., 1999). In erythroid progenitors the normal expression of Spi-1 is low and its deregulated overexpression results in erythroleukemia.
La leucemia mieloide crónica (LMC) es una de las leucemias más frecuentes. La LMC es una enfermedad bifásica. Primero cursa con una fase crónica, que suele durar 2-4 años. En esta fase la enfermedad es relativamente benigna, pero desemboca en todos los casos en una crisis blástica, que conduce a la muerte normalmente en menos de un año, a no ser que se someta al paciente a un transplante de médula ósea en su fase crónica. Esta fase es frecuentemente precedida de una fase acelerada, con características hematológicas intermedias entre la crónica y la blástica. Desde el punto de vista celular la fase crónica se caracteriza por un número elevado de granulocitos maduros, mientras que en la fase blástica se acumulan formas blásticas, normalmente de serie mieloide, y con menor frecuencia, linfoide. Junto con la linfocítica crónica es la leucemia más frecuente en gente mayor de 50 años, aunque de mucho peor pronóstico EL marcador molecular de la LMC es la proteína Bcr-Abl. Es una proteína de fusión con actividad proteína-tirosina quinasa desregulada, debido a una traslocación que yuxtapone los cromosomas 19 y 22 y da a lugar al cromosoma Filadelfia, marcador citogenética de la LMC (Deininger et al., 2000; Faderi et al., 1999; Goldman and Melo, 2003). El Bcr-Abl está ya presente en la fase crónica, y los mecanismos que gobiernan la transición de la fase crónica a la crisis blástica son desconocidos. Hasta ahora no se ha identificado un cambio molecular común o mayoritario en las crisis blásticas respecto a la fase crónica.Chronic myeloid leukemia (CML) is one of the most frequent leukemia. CML is a biphasic disease. First, it has a chronic phase , which usually lasts 2-4 years. In this phase the disease is relatively benign, but in all cases it leads to a blast crisis , which usually leads to death in less than a year, unless the patient is subjected to a bone marrow transplant in its chronic phase. . This phase is often preceded by an accelerated phase, with intermediate hematological characteristics between chronic and blast. From a cellular point of view, the chronic phase is characterized by a large number of mature granulocytes, while blast forms accumulate in the blast phase, usually myeloid series, and less frequently, lymphoid. Along with chronic lymphocytic is the most frequent leukemia in people over 50 years, although the molecular marker of CML is much worse prognosis is the Bcr-Abl protein. It is a fusion protein with deregulated protein-tyrosine kinase activity, due to a translocation that juxtaposes chromosomes 19 and 22 and gives rise to the Philadelphia chromosome, cytogenetic marker of CML (Deininger et al ., 2000; Faderi et al ., 1999; Goldman and Melo, 2003). Bcr-Abl is already present in the chronic phase, and the mechanisms that govern the transition from the chronic phase to the blast crisis are unknown. Until now, no common or major molecular change has been identified in blast crises compared to the chronic phase.
Históricamente, la LMC se ha tratado con busulfán e hidroxiurea, desde los años 80, con interferón \alpha, que suelen inducir remisión hematológica (Silver et al., 1999). Más recientemente se ha introducido el imatinib, un inhibidor de la quinasa Bcr-Abl. Los excelentes resultados obtenidos le están convirtiendo en el fármaco de primera línea en el tratamiento de la LMC (Kantarjian et al., 2002).Historically, CML has been treated with busulfan and hydroxyurea, since the 1980s, with interferon?, Which usually induce hematological remission (Silver et al ., 1999). More recently, imatinib, a Bcr-Abl kinase inhibitor, has been introduced. The excellent results obtained are making it the first-line drug in the treatment of CML (Kantarjian et al ., 2002).
Sin embargo, aunque estos fármacos, y especialmente el interferón y el imatinib pueden inducir remisiones hematológicas, citogenéticas y moleculares en la fase crónica, y el imatinib incluso en fase acelerada (Talpaz et al., 2002) y blástica (Druker et al., 2001; Sawyers et al., 2002), el único tratamiento curativo de la LMC hoy conocido es el trasplante alogénico de médula ósea en su fase crónica (Goldman and Melo, 2003). Se ha demostrado también que la eficacia curativa del trasplante es menor cuanto más tiempo lleva el paciente en fase crónica, pero es difícil evaluar el tiempo que lleva un paciente en fase crónica cuando se le hace el diagnóstico por primera vez (Goldman and Druker, 2001).However, although these drugs, and especially interferon and imatinib, can induce hematological, cytogenetic and molecular remissions in the chronic phase, and imatinib even in accelerated phase (Talpaz et al ., 2002) and blast (Druker et al ., 2001; Sawyers et al ., 2002), the only curative treatment of CML known today is allogeneic bone marrow transplantation in its chronic phase (Goldman and Melo, 2003). It has also been shown that the healing efficacy of the transplant is less the longer the patient has been in the chronic phase, but it is difficult to assess the time it takes for a patient in the chronic phase when the diagnosis is made for the first time (Goldman and Druker, 2001 ).
Hasta el momento, el único marcador molecular informativo sobre la evolución de la enfermedad en ausencia de datos hematológicos y citogenéticas es la determinación de la presencia del mRNA de Bcr-Abl. Esta determinación se hace por RT-PCR, usando cebadores que mapean en los genes BCR y ABL. Aunque la detección de mRNA de Bcr-Abl es útil en la detección de enfermedad mínima residual, hasta el momento no hay marcadores moleculares de la respuesta al tratamiento en LMC.So far, the only informative molecular marker on the evolution of the disease in the absence of hematological and cytogenetic data is the determination of the presence of the Bcr-Abl mRNA. This determination is made by RT-PCR, using primers that map in the BCR and ABL genes. Although the detection of Bcr-Abl mRNA is useful in the detection of minimal residual disease, so far there are no molecular markers of the response to treatment in CML.
Los inventores habían demostrado y publicado anteriormente que la expresión de SPI-1/PU.1 aumentaba al tratar células de una línea celular derivada de LMC (K562) con con interferón-\alpha, (Gutierrez et al., 1997). Posteriormente, los inventores han demostrado que la expresión de SPI-1/PU.1 se correlaciona con una recuperación hematológica de pacientes de LMC tratados con interferón-a e imatinib (resultados no publicados). Medida por RT-PCR, la expresión de SPI-1/PU.1 es baja al diagnóstico y aumenta durante al tratamiento con interferón o con imatinib, concomitante con la normalización hematológica. En la figura 1 se presenta una muestra representativa de la expresión de SPI-1/PU.1 en varios pacientes de LMC a lo largo de su tratamiento con interferón. Se obtuvieron resultados similares en 13 de 16 casos analizados. En los cuatro casos de la figura, el tratamiento indujo la recuperación de la hematopoyesis normal. En estos ensayos se usó el gen de la proteína ribosomal S14 como gen "housekeeping" como control de la cantidad de RNA y cDNA de las muestras. Este resultado es consistente con lo descrito previamente por los inventores sobre que la sobreexpresión de SPI-1/PU.1 en células humanas K562, provoca parada proliferativa, inducción de la diferenciación monocítica y bloqueo de la diferenciación eritroide (Gutierrez et al., 1997).The inventors had previously demonstrated and published that the expression of SPI-1 / PU.1 increased by treating cells of a cell line derived from LMC (K562) with interferon-α, (Gutierrez et al ., 1997). Subsequently, the inventors have demonstrated that the expression of SPI-1 / PU.1 correlates with a hematological recovery of CML patients treated with interferon-a and imatinib (unpublished results). Measured by RT-PCR, the expression of SPI-1 / PU.1 is low at diagnosis and increases during treatment with interferon or imatinib, concomitant with hematological normalization. A representative sample of the expression of SPI-1 / PU.1 in several CML patients throughout their treatment with interferon is presented in Figure 1. Similar results were obtained in 13 of 16 cases analyzed. In the four cases in the figure, the treatment induced the recovery of normal hematopoiesis. In these tests, the S14 ribosomal protein gene was used as a housekeeping gene as a control of the amount of RNA and cDNA in the samples. This result is consistent with what was previously described by the inventors that the overexpression of SPI-1 / PU.1 in human K562 cells causes proliferative arrest, induction of monocytic differentiation and blocking of erythroid differentiation (Gutierrez et al ., 1997 ).
Cuando se comparan los niveles de SPI-1/PU.1 (medidos por RT-PCR cuantitativa) en médula ósea de sujetos sanos, de pacientes de LMC al diagnóstico, y pacientes en fase crónica tratados con interferón y con imatinib se encontró que los niveles de mRNA de SPI-1/PU.1 eran eran mayores en los pacientes tratados. En la Figura 2 se resumen estas observaciones en 10 muestras de sujetos sanos, 31 de pacientes de LMC al diagnóstico, 32 tratados con interferón-\alpha y 17 tratados con imatinib (varios de ellos con tratamiento previo con interferón-\alpha.When the levels of SPI-1 / PU.1 (measured by quantitative RT-PCR) in bone marrow of healthy subjects, of CML patients at diagnosis, and chronic phase patients treated with interferon and imatinib were found that SPI-1 / PU.1 mRNA levels were higher in treated patients. Figure 2 summarizes these observations in 10 samples of healthy subjects, 31 of CML patients at diagnosis, 32 treated with interferon-α and 17 treated with imatinib (several of them with prior treatment with interferon-α.
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Deininger, M. W., Goldman, J. M., and Melo, J. V. (2000). The molecular biology of chronic myeloid leukemia. Blood 96, 3343-3356. Deininger , MW, Goldman , JM, and Melo , JV ( 2000 ). The molecular biology of chronic myeloid leukemia. Blood 96 , 3343-3356.
Druker, B. J., Sawyers, C. L., Kantarjian, H., Resta, D. J., Reese, S. F., Ford, J. M., Capdeville, R., and Talpaz, M. (2001). Activity of a specific inhibitor of the BCR-ABL tyrosine kinase in the blast crisis of chronic myeloid leukemia and acute lymphoblastic leukemia with the Philadelphia chromosome. N Engl J Med 344, 1038-1042. Druker , BJ, Sawyers , CL, Kantarjian , H., Resta , DJ, Reese , SF, Ford , JM, Capdeville , R., and Talpaz , M. ( 2001 ). Activity of a specific inhibitor of the BCR-ABL tyrosine kinase in the blast crisis of chronic myeloid leukemia and acute lymphoblastic leukemia with the Philadelphia chromosome. N Engl J Med 344 , 1038-1042.
Faders, S., Talpaz, M., Estrov, Z., and Kantarjian, H. M. (1999). Chronic myelogenous leukemia: biology and therapy. Ann Intern Med 131, 207-219. Faders , S., Talpaz , M., Estrov , Z., and Kantarjian , HM ( 1999 ). Chronic myelogenous leukemia: biology and therapy. Ann Intern Med 131 , 207-219.
Friedman, A. D. (2002). Transcriptional regulation of granulocyte and monocyte development. Oncogene 21, 3377-3390. Friedman , AD ( 2002 ). Transcriptional regulation of granulocyte and monocyte development. Oncogene 21 , 3377-3390.
Goldman, J. M., and Druker, B. J. (2001). Chronic myeloid leukemia: current treatment options. Blood 98, 2039-2042. Goldman , JM, and Druker , BJ ( 2001 ). Chronic myeloid leukemia: current treatment options. Blood 98 , 2039-2042.
Goldman, J. M., and Melo, J. V. (2003). Chronic myeloid leukemia--advances in biology and new approaches to treatment. N Engl J Med 349, 1451-1464. Goldman , JM, and Melo , JV ( 2003 ). Chronic myeloid leukemia - advances in biology and new approaches to treatment. N Engl J Med 349 , 1451-1464.
Gutierrez, P., Delgado, M. D., Richard, C., Moreau-Gachelin, F., and Leon, J. (1997). Interferon induces up-regulation of Spi-1/PU.1 in human leukemia K562 cells. Biochem Biophys Res Commun 240, 862-868. Gutierrez , P., Delgado , MD, Richard , C., Moreau-Gachelin , F., and Leon , J. ( 1997 ). Interferon induces up-regulation of Spi-1 / PU.1 in human leukemia K562 cells. Biochem Biophys Res Commun 240 , 862-868.
Kantarjian, H., Sawyers, C., Hochhaus, A., Guilhot, F., Schiffer, C., Gambacorti-Passerini, C., Niederwieser, D., Resta, D., Capdeville, R., Zoeliner, U., et al. (2002). Hematologic and cytogenetic responses to imatinib mesylate in chronic myelogenous leukemia. N Engl J Med 346, 645-652. Kantarjian , H., Sawyers , C., Hochhaus , A., Guilhot , F., Schiffer , C., Gambacorti-Passerini , C., Niederwieser , D., Resta , D., Capdeville , R., Zoeliner , U ., et al . ( 2002 ). Hematologic and cytogenetic responses to imatinib mesylate in chronic myelogenous leukemia. N Engl J Med 346 , 645-652.
Lloberas, J., Soler, C., and Celada, A. (1999). The key role of PU.1/SPI-1 in B cells, myeloid cells and macrophages. Immunol Today 20, 184-189. Lloberas , J., Soler , C., and Celada , A. ( 1999 ). The key role of PU.1 / SPI-1 in B cells, myeloid cells and macrophages. Immunol Today 20 , 184-189.
Sawyers, C. L., Hochhaus, A., Feldman, E., Goldman, J. M., Miller, C. B., Ottmann, O. G., Schiffer, C. A., Talpaz, M., Guilhot, F., Deininger, M. W., et al. (2002). Imatinib induces hematologic and cytogenetic responses in patients with chronic myelogenous leukemia in myeloid blast crisis: results of a phase II study. Blood 99, 3530-3539. Sawyers , CL, Hochhaus , A., Feldman , E., Goldman , JM, Miller , CB, Ottmann , OG, Schiffer , CA, Talpaz , M., Guilhot , F., Deininger , MW, et al . ( 2002 ). Imatinib induces hematologic and cytogenetic responses in patients with chronic myelogenous leukemia in myeloid blast crisis: results of a phase II study. Blood 99 , 3530-3539.
Silver, R. T., Woolf, S. H., Hehlmann, R., Appelbaum, F. R., Anderson, J., Bennett, C., Goldman, J. M., Guilhot, F., Kantarjian, H. M., Lichtin, A. E., et al. (1999). An evidence-based analysis of the effect of busulfan, hydroxyurea, interferon, and allogeneic bone marrow transplantation in treating the chronic phase of chronic myeloid leukemia: developed for the American Society of Hematology. Blood 94, 1517-1536. Silver , RT, Woolf , SH, Hehlmann , R., Appelbaum , FR, Anderson , J., Bennett , C., Goldman , JM, Guilhot , F., Kantarjian , HM, Lichtin , AE, et al . ( 1999 ). An evidence-based analysis of the effect of busulfan, hydroxyurea, interferon, and allogeneic bone marrow transplantation in treating the chronic phase of chronic myeloid leukemia: developed for the American Society of Hematology. Blood 94 , 1517-1536.
Talpaz, M., Silver, R. T., Druker, B. J., Goldman, J. M., Gambacorti-Passerini, C., Guilhot, F., Schiffer, C. A., Fischer, T., Deininger, M. W., Lennard, A. L., et al. (2002). Imatinib induces durable hematologic and cytogenetic responses in patients with accelerated phase chronic myeloid leukemia: results of a phase 2 study. Blood 99, 1928-1937. Talpaz , M., Silver , RT, Druker , BJ, Goldman , JM, Gambacorti-Passerini , C., Guilhot , F., Schiffer , CA, Fischer , T., Deininger , MW, Lennard , AL, et al . ( 2002 ). Imatinib induces durable hematologic and cytogenetic responses in patients with accelerated phase chronic myeloid leukemia: results of a phase 2 study. Blood 99 , 1928-1937.
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Los inventores han observado que la expresión del gen humano SPI-1/PU.1 es alta en células de médula ósea de sujetos sanos pero es significativamente más baja en muestras de pacientes de LMC al diagnóstico en fase crónica. Sin embargo, en pacientes de LMC que están en remisión hematológica, la expresión de SPI-1/PU.1 se recupera hasta alcanzar valores normales. Esta regulación positiva de SPI-1/PU.1 se ha observado tanto en pacientes de LMC tratados con interferón-\alpha y con imatinib (los dos tratamientos hoy en uso para esta leucemia). El procedimiento que se presenta consiste en medir la expresión de SPI-1/PU.1 en muestras de médula ósea de pacientes y referirla a una muestra de referencia que exprese dicho gen. Esta referencia puede ser médula ósea de sujetos sanos o de una línea celular que exprese dicho gen, como K562 tratadas con interferón. Una expresión alta o recuperada de SPI-1/PU.1 es indicador de restablecimiento de hematopoyesis normal y/o una baja proporción de células leucémicas en la muestra considerada, y por tanto, una indicación de éxito del tratamiento.The inventors have observed that the expression of the human SPI-1 / PU.1 gene is high in bone marrow cells of healthy subjects but is significantly lower in samples of CML patients at chronic phase diagnosis. However, in CML patients who are in hematological remission, the expression of SPI-1 / PU.1 is recovered until normal values are reached. This positive regulation of SPI-1 / PU.1 has been observed both in CML patients treated with interferon-α and with imatinib (the two treatments currently used for this leukemia). The procedure presented is to measure the expression of SPI-1 / PU.1 in bone marrow samples of patients and refer it to a reference sample that expresses said gene. This reference may be bone marrow of healthy subjects or a cell line that expresses said gene, such as K562 treated with interferon. A high or recovered expression of SPI-1 / PU.1 is an indicator of restoration of normal hematopoiesis and / or a low proportion of leukemic cells in the sample considered, and therefore, an indication of treatment success.
Figura 1. Expresión de SPI-1/PU.1 durante la evolución de LMC. Las muestras 1.1., 2.1, 3.1 y 4.1 corresponden al diagnóstico. Las siguientes son muestras de los pacientes 1, 2, 3 y 4 tomadas con durante el tratamiento con interferón-\alpha. En todos los casos los pacientes entraron en remisión hematológica. Se observa como la expresión de SPI-1/PU.1 aumenta según se va consiguiendo (mediante el tratamiento) la normalización hematopoyética. A la derecha se indica entre paréntesis el tamaño de los amplicones de SPI-1/PU.1 y de S14. A la izquierda los tamaños de marcadores de tamaño de DNA.Figure 1. SPI-1 / PU.1 expression during LMC evolution. Samples 1.1., 2.1, 3.1 and 4.1 correspond to the diagnosis. The following are samples of patients 1, 2, 3 and 4 taken with during interferon-α treatment. In all cases the patients went into hematological remission. It is observed how the expression of SPI-1 / PU.1 increases as hematopoietic normalization is achieved (by treatment). On the right, the size of the SPI-1 / PU.1 and S14 amplicons is indicated in parentheses. On the left the sizes of DNA size markers.
Figura 2. Expresión de SPI-1/PU.1 (analizada por RT-PCR cuantitativa y normalizada respecto a la de S14) en muestras de sujetos sanos (control), y pacientes de LMC al diagnóstico y tratados in interferón-a e imatinib. En cada caso se indica el número de muestras analizadas. Las barras de error indican el Error Estándar de la Media.Figure 2. Expression of SPI-1 / PU.1 (analyzed by RT-PCR quantitative and normalized with respect to that of S14) in samples of healthy subjects (control), and CML patients at diagnosis and treated in interferon-a and imatinib. In each case it Indicates the number of samples analyzed. Error bars indicate the Standard Error of the Average.
Se extraerá RNA de células mononucleares separadas de médula ósea o de sangre periférica. El RNA se copia a cDNA usando transcriptasa inversa y cebadores al azar ("random hexamers") o bien oligonucleótidos específicos de regiones exónicas del gen SPI-1/PU.1 humano. El cDNA es amplificado usando como cebadores ("primers") dos oligonucleótidos que mapean en secuencias exónicas del gen SPI-1/PU.1 humano. El resultado del PCR se puede evaluar analizando los productos de la reacción de PCR por electroforesis en gel de agarosa o bien por PCR cuantitativa o a tiempo real.RNA will be extracted from mononuclear cells separated from bone marrow or peripheral blood. RNA is copied to cDNA using reverse transcriptase and random primers ("random hexamers") or oligonucleotides specific to exonic regions of the human SPI-1 / PU.1 gene. The cDNA is amplified using as primers ("primers") two oligonucleotides that map into exonic sequences of the human SPI-1 / PU.1 gene. The PCR result can be evaluated by analyzing the products of the PCR reaction by agarose gel electrophoresis or by quantitative or real-time PCR.
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| US20030119043A1 (en) * | 2001-11-09 | 2003-06-26 | Tanner Stephan Markus | BAALC expression as a diagnostic marker for acute leukemia |
| WO2004031409A2 (en) * | 2002-09-30 | 2004-04-15 | Oncotherapy Science, Inc. | Method for diagnosing chronic myeloid leukemia |
| US20040161760A1 (en) * | 2001-03-01 | 2004-08-19 | Hiroyuki Mano | Method of molecular diagnosis of chronic myelogenous leukemia |
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2004
- 2004-11-22 ES ES200402864A patent/ES2315040B2/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992021032A1 (en) * | 1991-05-24 | 1992-11-26 | The Regents Of The University Of California | Methods for the detection of bcr-abl and abnormal abl proteins in leukemia patients |
| US20040161760A1 (en) * | 2001-03-01 | 2004-08-19 | Hiroyuki Mano | Method of molecular diagnosis of chronic myelogenous leukemia |
| US20030119043A1 (en) * | 2001-11-09 | 2003-06-26 | Tanner Stephan Markus | BAALC expression as a diagnostic marker for acute leukemia |
| WO2004031409A2 (en) * | 2002-09-30 | 2004-04-15 | Oncotherapy Science, Inc. | Method for diagnosing chronic myeloid leukemia |
Non-Patent Citations (3)
| Title |
|---|
| DELGADO MD., et al., "{}Spi-1/PU.1 proto-oncogene induces opposite effects on monocytic and erythroid differentation of K562 cells"{} Biochememical and Biophysical Research Communications (1998), 252, páginas 383-391. * |
| GUTIERREZ P., et al., "{}Interferon induces up-regulation of Spi-1/ PU.1 in human leukemia K562 cells"{} Biochememical and Biophysical Research Communications (1997), 240, páginas 862-868. Citado en la solicitud. * |
| LLOBERAS J., et al., "{}The key role of PU.1/Spi-1 in B cells, myeloid cells and macrophages"{} Immunology Today (1999), 20(4), páginas 184-189. Citado en la solicitud. * |
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