Estudios genéticos en linfomas cutáneos de células T

Mycosis fungoides (MF) is a clonally derived lymphoproliferative disorder that\npreferentially involves the skin, which etiology and pathogenesis remains elusive.\nMorphological and clinical features are not always accurate enough to predict the disease\noutcome. Comparative genomic hybridization an...

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Autor principal:	Huamán Garaicoa, Fuad Olmedo
Otros Autores:	Slavutsky, Irma
Formato:	Tesis de maestría acceptedVersion
Lenguaje:	Español
Publicado:	Facultad de Farmacia y Bioquímica 2016
Materias:	Linfomas cutáneos T Fish Microrna Expresión génica Desbalances genómicos 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_1915 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_1915.dir/1915.PDF
Aporte de:	Repositorio Digital de la Universidad de Buenos Aires (UBA) de Universidad de Buenos Aires

id	I28-R145-HWA_1915
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	Linfomas cutáneos T Fish Microrna Expresión génica Desbalances genómicos Ciencias de la vida
spellingShingle	Linfomas cutáneos T Fish Microrna Expresión génica Desbalances genómicos Ciencias de la vida Huamán Garaicoa, Fuad Olmedo Estudios genéticos en linfomas cutáneos de células T
topic_facet	Linfomas cutáneos T Fish Microrna Expresión génica Desbalances genómicos Ciencias de la vida
description	Mycosis fungoides (MF) is a clonally derived lymphoproliferative disorder that\npreferentially involves the skin, which etiology and pathogenesis remains elusive.\nMorphological and clinical features are not always accurate enough to predict the disease\noutcome. Comparative genomic hybridization and microarrays have shown imbalances of\nCDKN2A (9p21) and C-MYC (8q24), with probable prognostic value in this pathology. In\naddition, miR-155 overexpression was observed in several hematological and solid tumors,\npromoting genomic instability, proliferation, and survival of malignant cells. In this study,\nwe have evaluated CDKN2A losses and C-MYC gains and their association with miR-155\nexpression, in patients with diagnosis of MF. Results were correlated with\nclinicopathological features of patients. The study was performed on formalin-fixed and\nparaffin-embedded biopsies from 36 patients with MF (26 males; median age 62.5 years,\nrange: 31-82 years): 16 cases were tumor stage MF (T-MF), 13 were MF with histological\ntransformation to a large T-cell lymphoma (TR-MF) and 7 were folliculotropic MF (F-MF)\nvariant. FISH analysis using OTS9P21.3 (CDKN2A) and OTS8Q24 (C-MYC) probes\n(LiVE-Lexel, Argentina), was performed. Gene expression was quantified by real time\nPCR using TaqMan methodology. Ten control samples from benign skin diseases were also\nevaluated. The study was approved by the local Ethics Committee. All individuals provided\ntheir informed written consent. FISH study was performed in 34 patients, 20 (59%) showed\ngenomic alterations (GA): 8 (40%) cases had CDKN2A deletion, 7 (35%) showed C-MYC\ngains and 5 (25%) exhibited both anomalies. CDKN2A deletion was observed in 7/13\n(54%) TR-MF, 4/7 (57%) F-MF and 2/16 (12.5%) T-MF (p=0.03); meanwhile C-MYC gain\nwas detected in 7/13 (54%) TR-MF and 5/7 (71.4%) F-MF. Thus, GA rate was lower in TMF\n14.3% with respect to TR-MF 92.3% and F-MF 85.7% (p=0.001 and p=0.004,\nrespectively). These aberrations were more frequent in head, neck and lower limbs (77.8%)\nin contrast with trunk and upper limbs (40%) (p=0.03). By immunohistochemistry, majority\nof cases with <25% CD30+ cells did not show GA (91% vs 9%) (p=0.01); meanwhile\npatients of GA group had more high median proliferation index (Ki-67) (49.5%) in\ncomparison to cases with no genomic aberrations (NA) (5%) (p=0.003). Interestingly, TRMF\nwith follicular origin had more frequently 9p21 deletion (80%) and less 8q24 gain\n(40%) respect to classic origin (42.8% and 71.4%, respectively). Clinical response to treatment was absent in 11/20 (55%) of cases with GA vs. 2/14 (14.3%) patients with NA\n(p=0.02). Although no significant differences were reached, mean LDH and Beta 2\nmicroglobulin levels and extracutaneous relapse were higher with respect to those with NA.\nIn addition, the group with GA showed shorter overall survival (92 months) compared to\ncases with NA, that not reached the median survival (Log-rank p=0.04), indicating their\nassociation with poor outcome. Gene expression analysis (n=36) showed miR-155\nupregulation in 25% TR-MF and 28.6% F-MF and in only 7.7% T-MF. No miR-155\nexpression was observed in controls. The correlation with FISH results found miR-155\noverexpression in 26.3% of patients with GA and 8.3% of cases with NA. Our results\nshowed high proportion of 9p21 losses and 8q24 gains in MF patients. TR-MF exhibited\nthe highest frequency of GA, supporting a role in the process of neoplastic transformation.\nAlthough the number of F-MF is reduced in our series, this morphological variant would\nseem to be associated to an increased frequency of genomic imbalances. Moreover, our\nfindings on miR-155 expression support its relation with genomic instability and tumor\ndevelopment, contributing a better biological characterization of this pathology.
author2	Slavutsky, Irma
author_facet	Slavutsky, Irma Huamán Garaicoa, Fuad Olmedo
format	Tesis de maestría Tesis de maestría acceptedVersion
author	Huamán Garaicoa, Fuad Olmedo
author_sort	Huamán Garaicoa, Fuad Olmedo
title	Estudios genéticos en linfomas cutáneos de células T
title_short	Estudios genéticos en linfomas cutáneos de células T
title_full	Estudios genéticos en linfomas cutáneos de células T
title_fullStr	Estudios genéticos en linfomas cutáneos de células T
title_full_unstemmed	Estudios genéticos en linfomas cutáneos de células T
title_sort	estudios genéticos en linfomas cutáneos de células t
publisher	Facultad de Farmacia y Bioquímica
publishDate	2016
url	http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_1915 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_1915.dir/1915.PDF
work_keys_str_mv	AT huamangaraicoafuadolmedo estudiosgeneticosenlinfomascutaneosdecelulast
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spelling	I28-R145-HWA_19152019-09-27 Mycosis fungoides (MF) is a clonally derived lymphoproliferative disorder that\npreferentially involves the skin, which etiology and pathogenesis remains elusive.\nMorphological and clinical features are not always accurate enough to predict the disease\noutcome. Comparative genomic hybridization and microarrays have shown imbalances of\nCDKN2A (9p21) and C-MYC (8q24), with probable prognostic value in this pathology. In\naddition, miR-155 overexpression was observed in several hematological and solid tumors,\npromoting genomic instability, proliferation, and survival of malignant cells. In this study,\nwe have evaluated CDKN2A losses and C-MYC gains and their association with miR-155\nexpression, in patients with diagnosis of MF. Results were correlated with\nclinicopathological features of patients. The study was performed on formalin-fixed and\nparaffin-embedded biopsies from 36 patients with MF (26 males; median age 62.5 years,\nrange: 31-82 years): 16 cases were tumor stage MF (T-MF), 13 were MF with histological\ntransformation to a large T-cell lymphoma (TR-MF) and 7 were folliculotropic MF (F-MF)\nvariant. FISH analysis using OTS9P21.3 (CDKN2A) and OTS8Q24 (C-MYC) probes\n(LiVE-Lexel, Argentina), was performed. Gene expression was quantified by real time\nPCR using TaqMan methodology. Ten control samples from benign skin diseases were also\nevaluated. The study was approved by the local Ethics Committee. All individuals provided\ntheir informed written consent. FISH study was performed in 34 patients, 20 (59%) showed\ngenomic alterations (GA): 8 (40%) cases had CDKN2A deletion, 7 (35%) showed C-MYC\ngains and 5 (25%) exhibited both anomalies. CDKN2A deletion was observed in 7/13\n(54%) TR-MF, 4/7 (57%) F-MF and 2/16 (12.5%) T-MF (p=0.03); meanwhile C-MYC gain\nwas detected in 7/13 (54%) TR-MF and 5/7 (71.4%) F-MF. Thus, GA rate was lower in TMF\n14.3% with respect to TR-MF 92.3% and F-MF 85.7% (p=0.001 and p=0.004,\nrespectively). These aberrations were more frequent in head, neck and lower limbs (77.8%)\nin contrast with trunk and upper limbs (40%) (p=0.03). By immunohistochemistry, majority\nof cases with <25% CD30+ cells did not show GA (91% vs 9%) (p=0.01); meanwhile\npatients of GA group had more high median proliferation index (Ki-67) (49.5%) in\ncomparison to cases with no genomic aberrations (NA) (5%) (p=0.003). Interestingly, TRMF\nwith follicular origin had more frequently 9p21 deletion (80%) and less 8q24 gain\n(40%) respect to classic origin (42.8% and 71.4%, respectively). Clinical response to treatment was absent in 11/20 (55%) of cases with GA vs. 2/14 (14.3%) patients with NA\n(p=0.02). Although no significant differences were reached, mean LDH and Beta 2\nmicroglobulin levels and extracutaneous relapse were higher with respect to those with NA.\nIn addition, the group with GA showed shorter overall survival (92 months) compared to\ncases with NA, that not reached the median survival (Log-rank p=0.04), indicating their\nassociation with poor outcome. Gene expression analysis (n=36) showed miR-155\nupregulation in 25% TR-MF and 28.6% F-MF and in only 7.7% T-MF. No miR-155\nexpression was observed in controls. The correlation with FISH results found miR-155\noverexpression in 26.3% of patients with GA and 8.3% of cases with NA. Our results\nshowed high proportion of 9p21 losses and 8q24 gains in MF patients. TR-MF exhibited\nthe highest frequency of GA, supporting a role in the process of neoplastic transformation.\nAlthough the number of F-MF is reduced in our series, this morphological variant would\nseem to be associated to an increased frequency of genomic imbalances. Moreover, our\nfindings on miR-155 expression support its relation with genomic instability and tumor\ndevelopment, contributing a better biological characterization of this pathology. Fil: Huamán Garaicoa, Fuad Olmedo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Buenos Aires, Argentina Facultad de Farmacia y Bioquímica Slavutsky, Irma Huamán Garaicoa, Fuad Olmedo 2016-05-06 La micosis fungoide (MF) es un tipo de desorden linfoproliferativo clonal que\npreferentemente involucra la piel, cuya etiología y patogénesis son inciertas. Las\ncaracterísticas clínicas y morfológicas no siempre resultan suficientes para predecir el\npronóstico de esta enfermedad. Las técnicas de hibridación genómica comparada y\nmicroarrays han mostrado desbalances de los genes CDKN2A (9p21) y C-MYC (8q24), con\nun probable valor pronóstico en esta patología. Además la sobreexpresión de miR-155 fue\nobservada en diferentes tumores sólidos y hematológicos, promoviendo la inestabilidad\ngenómica, proliferación y supervivencia de las células malignas. En el presente estudio,\nevaluamos los desbalances genómicos de CDKN2A y C-MYC, así como su asociación con\nla expresión de miR-155 en pacientes con diagnóstico de MF. Los resultados fueron\ncorrelacionados con las características clínico-patológicas de los pacientes. El estudio fue\nrealizado en biopsias fijadas en formol y embebidas en parafina de 36 pacientes con MF\n(26 varones; edad media 62,5 años, rango: 31-82 años): 16 casos correspondieron a MF\nestadio tumoral (MF-T), 13 a MF con transformación histológica a un linfoma T de células\ngrandes (MF-TR), y 7 casos a la variante MF foliculotrópica (MF-F). El análisis de FISH se\nrealizó usando las sondas OTS9P21.3 (CDKN2A) y OTS8Q24 (C-MYC) (LiVE-Lexel,\nArgentina). Se cuantificó la expresión de miR-155 mediante PCR en tiempo real usando\nmetodología TaqMan. Se evaluaron además diez muestras de enfermedades inflamatorias\ncutáneas, usadas como controles. El estudio fue aprobado por el Comité de Ética local.\nTodos los individuos proveyeron consentimiento informado para su participación. El\nestudio de FISH fue realizado en 34 pacientes, 20 (59%) mostraron alteraciones genómicas\n(AG): 8 (40%) casos tuvieron deleción de CDKN2A, 7 (35%) mostraron ganancias de CMYC\ny 5 (25%) exhibieron ambos desbalances. La deleción de CDKN2A fue observada en\n7/13 (54%) MF-TR, 4/7 (57%) MF-F y 2/16 (12,5%) MF-T (p=0,03); mientras que la\nganancia de C-MYC fue detectada en 7/13 (54%) MF-TR y en 5/7 (71,4%) MF-F. De esta\nforma, la frecuencia de AG fue de 14,3% MF-T frente a 92,3% MF-TR y 85,7% MF-F\n(p=0,001 y p=0,004, respectivamente). Estas aberraciones fueron más frecuentes en las\nlesiones ubicadas en cabeza, cuello y extremidades inferiores (77,8%) en contraste con las\nhalladas en el tronco y miembros superiores (40%) (p=0,03). Por inmunohistoquímica, la\nmayoría de los casos con < 25% células CD30+ no presentaban AG (91% vs 9%) (p=0,01),\nmientras que los pacientes con alteraciones mostraron una mayor media del índice de\nproliferación (Ki-67) (49,5%) que los casos sin alteraciones (SA) (5%) (p=0,003).\nAdicionalmente, las MF-TR de origen foliculotropo tenían más frecuentemente deleción\n9p21 (80%) que ganancia de 8q24 (40%) respecto a las de origen clásico (42,8% y 71,4%,\nrespectivamente). La respuesta clínica al tratamiento fue ausente en 11/20 (55%) de los\ncasos con AG respecto a 2/14 (14,3%) pacientes SA (p=0,02). Aunque no se alcanzaron\ndiferencias significativas, los niveles de LDH y Beta 2 microglobulina, y la recaída\nextracutánea fueron mayores en relación al grupo SA. Además, el grupo con AG mostró la\nsobrevida más corta (92 meses) comparado con los casos SA, que no alcanzaron la media\nde supervivencia (Log-rank p=0,04), indicando su asociación con un pronóstico\ndesfavorable. El análisis de la expresión génica (n=36) mostró sobreexpresión de miR-155\nen el 25% de las MF-TR y en el 28,6% de las MF-F, mientras que sólo se observó en el\n7,7% 7,7% de las MF-T. No se detectó sobreexpresión de miR-155 en los controles. La\ncorrelación con los resultados de FISH mostró incremento de la expresión de miR-155 en el\n26,3% de los pacientes con AG respecto de 8,3% de los casos SA. Nuestros resultados\nmuestran una alta proporción de pérdidas de 9p21 y 8q24 en pacientes con MF. La MF-TR\nexhibió la mayor frecuencia de AG, apoyando un rol de éstas alteraciones en el proceso de\ntransformación neoplásica. Aún cuando el número de MF-F es reducido en nuestra serie,\nesta variante morfológica podría estar asociada a una mayor frecuencia de AG. Finalmente,\nnuestros hallazgos en la expresión de miR-155 apoyan su relación con la inestabilidad\ngenómica y el desarrollo tumoral, pudiendo constituir un aporte para la profundización de\nla caracterización biológica de esta patología. application/pdf Wainstok, Rosa Vanzulli, Silvia Rossetti, Liliana Linfomas cutáneos T Fish Microrna Expresión génica Desbalances genómicos 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 Estudios genéticos en linfomas cutáneos de células T 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_1915 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_1915.dir/1915.PDF

Estudios genéticos en linfomas cutáneos de células T

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