Cuantificación del transcripto BCR-ABL1 relativo a diferentes genes control: ABL1 y GUSB en pacientes con leucemia mieloide crónica

To evaluate the expression levels of the BCR-ABL1 rearrangement in Chronic Myeloid Leukemia, it is used ABL1 as control gene. Taking into account that this gene is involved in the translocation, others genes have been postulated, including beta-glucuronidase (GUSB) that is located on the long arm of...

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Autor principal: Paggi D'Agostino, Andrés
Otros Autores: Bianchini, Michele
Formato: Tesis de maestría acceptedVersion
Lenguaje:Español
Publicado: Facultad de Farmacia y Biquímica 2017
Materias:
ABL1
Leucemia Mieloide Crónica
GUSB
Cáncer
Ciencias de la vida
Acceso en línea:http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_2064
http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_2064.dir/2064.PDF
Aporte de:
Repositorio Digital de la Universidad de Buenos Aires (UBA) de Universidad de Buenos Aires
id I28-R145-HWA_2064
record_format dspace
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-145
collection Repositorio Digital de la Universidad de Buenos Aires (UBA)
language Español
orig_language_str_mv spa
topic ABL1
Leucemia Mieloide Crónica
GUSB
Cáncer
Ciencias de la vida
spellingShingle ABL1
Leucemia Mieloide Crónica
GUSB
Cáncer
Ciencias de la vida
Paggi D'Agostino, Andrés
Cuantificación del transcripto BCR-ABL1 relativo a diferentes genes control: ABL1 y GUSB en pacientes con leucemia mieloide crónica
topic_facet ABL1
Leucemia Mieloide Crónica
GUSB
Cáncer
Ciencias de la vida
description To evaluate the expression levels of the BCR-ABL1 rearrangement in Chronic Myeloid Leukemia, it is used ABL1 as control gene. Taking into account that this gene is involved in the translocation, others genes have been postulated, including beta-glucuronidase (GUSB) that is located on the long arm of chromosome 7 and it is not involved in the BCR-ABL1 rearrangement. In the present study three parameters of early molecular response were evaluated: 1. Level of transcripts on an international scale at three months; 2.- Reduction of transcripts to achive half of the baseline value (Halving time) with both control genes; 3.- Logarithmic reduction at three months of treatment (using the GUSB control gene). Through the real-time polymerase chain reaction all involved transcripts (BCR-ABL1, ABL1 and GUSB) were quantified and a conversion factor for GUSB was calculated to validate the results on an international scale. Forty-four patient samples were studied at diagnosis, 18 at three months and 11 at six months of follow-up of the disease under treatment with tyrosine kinase inhibitors (TKI). Analyzing the means of transcripts-BCR-ABL1 at diagnosis, at 3 and 6 months, it was observed that at diagnosis the BCR-ABL1/GUSB ratio presented lower values respect to BCR-ABL1/ABL1, whereas at 3 and 6 months the values were coincident. A positive correlation was observed between the level of GUSB and BCR-ABL1 transcripts which could explain the results with GUSB at diagnosis. The decrease in BCR-ABL1 transcripts at 3 months and 6 months from the time of diagnosis was greater using the ABL1 control gene than with the GUSB control gene. The rate of reduction of BCR-ABL1 transcripts (Halving time) was evaluated with both control\n7\ngenes. ROC curves were performed, obtaining cut off values of ? 26 days for GUSB and ? 23 days for ABL1. With this analysis, in addition, it was observed that ABL1 has greater discrimination power than GUSB between responders and non-responders. Using the Fisher test applied to the data obtained with the ABL1 control gene, we observed that the Halving time ? 23 days was associated with a Major Molecular Response at 12 months of treatment. Our data indicate that ABL1 allows to discriminate with greater sensitivity the response to the treatment with TKIs compared to GUSB.
author2 Bianchini, Michele
author_facet Bianchini, Michele
Paggi D'Agostino, Andrés
format Tesis de maestría
Tesis de maestría
acceptedVersion
author Paggi D'Agostino, Andrés
author_sort Paggi D'Agostino, Andrés
title Cuantificación del transcripto BCR-ABL1 relativo a diferentes genes control: ABL1 y GUSB en pacientes con leucemia mieloide crónica
title_short Cuantificación del transcripto BCR-ABL1 relativo a diferentes genes control: ABL1 y GUSB en pacientes con leucemia mieloide crónica
title_full Cuantificación del transcripto BCR-ABL1 relativo a diferentes genes control: ABL1 y GUSB en pacientes con leucemia mieloide crónica
title_fullStr Cuantificación del transcripto BCR-ABL1 relativo a diferentes genes control: ABL1 y GUSB en pacientes con leucemia mieloide crónica
title_full_unstemmed Cuantificación del transcripto BCR-ABL1 relativo a diferentes genes control: ABL1 y GUSB en pacientes con leucemia mieloide crónica
title_sort cuantificación del transcripto bcr-abl1 relativo a diferentes genes control: abl1 y gusb en pacientes con leucemia mieloide crónica
publisher Facultad de Farmacia y Biquímica
publishDate 2017
url http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_2064
http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_2064.dir/2064.PDF
work_keys_str_mv AT paggidagostinoandres cuantificaciondeltranscriptobcrabl1relativoadiferentesgenescontrolabl1ygusbenpacientesconleucemiamieloidecronica
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spelling I28-R145-HWA_20642022-02-15 To evaluate the expression levels of the BCR-ABL1 rearrangement in Chronic Myeloid Leukemia, it is used ABL1 as control gene. Taking into account that this gene is involved in the translocation, others genes have been postulated, including beta-glucuronidase (GUSB) that is located on the long arm of chromosome 7 and it is not involved in the BCR-ABL1 rearrangement. In the present study three parameters of early molecular response were evaluated: 1. Level of transcripts on an international scale at three months; 2.- Reduction of transcripts to achive half of the baseline value (Halving time) with both control genes; 3.- Logarithmic reduction at three months of treatment (using the GUSB control gene). Through the real-time polymerase chain reaction all involved transcripts (BCR-ABL1, ABL1 and GUSB) were quantified and a conversion factor for GUSB was calculated to validate the results on an international scale. Forty-four patient samples were studied at diagnosis, 18 at three months and 11 at six months of follow-up of the disease under treatment with tyrosine kinase inhibitors (TKI). Analyzing the means of transcripts-BCR-ABL1 at diagnosis, at 3 and 6 months, it was observed that at diagnosis the BCR-ABL1/GUSB ratio presented lower values respect to BCR-ABL1/ABL1, whereas at 3 and 6 months the values were coincident. A positive correlation was observed between the level of GUSB and BCR-ABL1 transcripts which could explain the results with GUSB at diagnosis. The decrease in BCR-ABL1 transcripts at 3 months and 6 months from the time of diagnosis was greater using the ABL1 control gene than with the GUSB control gene. The rate of reduction of BCR-ABL1 transcripts (Halving time) was evaluated with both control\n7\ngenes. ROC curves were performed, obtaining cut off values of ? 26 days for GUSB and ? 23 days for ABL1. With this analysis, in addition, it was observed that ABL1 has greater discrimination power than GUSB between responders and non-responders. Using the Fisher test applied to the data obtained with the ABL1 control gene, we observed that the Halving time ? 23 days was associated with a Major Molecular Response at 12 months of treatment. Our data indicate that ABL1 allows to discriminate with greater sensitivity the response to the treatment with TKIs compared to GUSB. Fil: Piaggi D'Agostino, Andrés. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Buenos Aires, Argentina Bianchini, Michele Facultad de Farmacia y Biquímica Larripa, Irene Paggi D'Agostino, Andrés 2017-09-04 Para evaluar los niveles de expresión del reordenamiento BCR-ABL1 en la Leucemia Mieloide Crónica, se realiza la cuantificación de dichos transcriptos respecto del gen control ABL1. Debido a que este gen está involucrado en la translocación, se sugirieron otros, entre ellos, beta-glucuronidasa (GUSB) el cual se localiza en el brazo largo del cromosoma 7 y no está implicado en el rearreglo molecular BCR-ABL1. En el presente estudio se evaluaron tres parámetros de respuesta molecular temprana: 1.- Nivel de transcriptos en escala internacional a los tres meses de tratamiento; 2.- Reducción de los transcriptos a la mitad (Halving time) con ambos genes control; 3.- Reducción logarítmica a tres meses de tratamiento (utilizando al gen control GUSB).\nA través de la PCR en tiempo real se cuantificaron los transcriptos: BCR-ABL1, ABL1 y GUSB y se calculó el factor de conversión para este último utilizando los calibradores de la WHO para validar los resultados a la escala internacional. Se estudiaron 44 muestras de pacientes al diagnóstico, 18 a los tres meses y 11 a los seis meses de seguimiento de la enfermedad bajo tratamiento con inhibidores de tirosina kinasa (ITK). Analizando las medias poblacionales de transcriptos-BCR-ABL1 al diagnóstico, a los 3 y 6 meses se observó que al diagnóstico la relación BCR-ABL1/GUSB presentaba valores menores respecto de BCR-ABL1/ABL1, mientras que a los 3 y 6 meses los valores eran coincidentes. Se observó una correlación positiva entre el nivel de transcriptos GUSB y BCR-ABL1 lo cual podría explicar los resultados con GUSB al diagnóstico. La disminución de los transcriptos BCR-ABL1 a los 3 y 6 meses respecto del momento del diagnóstico fue mayor utilizando el gen control ABL1 que con el gen control GUSB. Se analizó la velocidad de reducción de los transcriptos BCR-ABL1 a la mitad (Halving time) con ambos genes control. Para ello se realizaron las curvas ROC obteniéndose valores de corte ? 26 días para GUSB y ? 23 días para ABL1. Con este análisis, además, se observó que ABL1 tiene mayor poder de discriminación que GUSB entre respondedores y no respondedores. Mediante el test de Fisher aplicado a los\n6\ndatos obtenidos con el gen control ABL1, observamos que la variable Halving time ? a 23 días se asoció con una Respuesta Molecular Mayor a los 12 meses de tratamiento (p=0.013). Nuestros datos indican que el ABL1 permite discriminar con mayor sensibilidad la respuesta al tratamiento con ITKs comparado con GUSB. application/pdf Carballo, Marta Ana Levy, Estrella Ferrer, Marcela ABL1 Leucemia Mieloide Crónica GUSB Cáncer spa Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ Ciencias de la vida Cuantificación del transcripto BCR-ABL1 relativo a diferentes genes control: ABL1 y GUSB en pacientes con leucemia mieloide crónica info:eu-repo/semantics/masterThesis info:ar-repo/semantics/tesis de maestría info:eu-repo/semantics/acceptedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_2064 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_2064.dir/2064.PDF

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