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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2019.tde-13122021-185707
Document
Author
Full name
José Carlos Mann Prado
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2019
Supervisor
Committee
Pierulivo, Enrique Mario Boccardo (President)
Durigon, Edison Luiz
Romano, Camila Malta
Tomimori, Jane
Title in Portuguese
Detecção e caracterização molecular de Poliomavírus de células de Merkel em carcinomas de células de Merkel.
Keywords in Portuguese
Câncer de pele
Sequenciamento
Vírus de DNA
Abstract in Portuguese
O carcinoma de células de Merkel (CCM) é uma neoplasia rara, muito agressiva que afeta principalmente indivíduos maiores de 50 anos e com o sistema imunológico comprometido. A associação do Poliomavírus humanos 5 (MCPyV) ao CCM foi estabelecida e é aceita até o momento como a única associação positiva entre um Poliomavírus e uma neoplasia em humanos. No entanto, dados como prevalência viral e presença de possíveis variantes moleculares específicas de uma determinada região geográfica não têm sido analisados em amostras do Brasil. No presente estudo, analisamos 84 amostras de 57 pacientes diagnosticados com CCM obtidas do AC Camargo Cancer Center, visando determinar: i) a prevalência de MCPyV; ii) as caracteristicas filogenéticas das sequências virais identificadas por sequenciamento; iii) a prevalência de co-infecção com HPV e iv) avaliar o padrão de expressão de proteínas relacionadas a perda de polaridade celular. Através dessa análise determinamos que a prevalência viral nas amostras estudadas é de 94,8%. Na caracterização das sequências geradas, analisarmos os fragmentos virais LT3 e VP1 e identificamos quatro variantes moleculares correspondentes a três variantes e a sequência protótipo em cada um destes. As variantes identificadas foram comparadas às descritas em banco de dados e suas sequências foram utilizadas para construir árvores filogenéticas. Dessa maneira, verificamos uma distribuição ampla e aleatória das variantes identificados nos ramos destas árvores. Finalmente, analisamos o padrão de expressão de proteínas relacionadas ao evento de perda de polaridade. De maneira geral, não observamos diferenças significates entre os padrões de marcação para as proteínas analisadas individualmente entre amostras positivas em negativas para o vírus.
Title in English
Detection and molecular characterization of Merkel cell Polyomavirus in Merkel cell carcinoma carcinomas de células de Merkel.
Keywords in English
DNA vírus
Sequencing
Skin Cancer
Abstract in English
Merkel cell carcinoma (MCC) is a rare, very aggressive neoplasm that mainly affects individuals older than 50 years and with compromised immune systems. The association of human Polyomavirus 5 (MCPyV) with CCM has been established and is accepted to date as the only positive association between a Polyomavirus and a neoplasm in humans. However, data such as viral prevalence and the presence of possible molecular variants specific to a given geographical region have not been analyzed in samples from Brazil. In the present study, we analyzed 84 samples from 57 patients diagnosed with CCM obtained from the AC Camargo Cancer Center, in order to determine: i) the prevalence of MCPyV; ii) the phylogenetic characteristics of the viral sequences identified by sequencing; iii) the prevalence of HPV co-infection and iv) to evaluate the expression pattern of proteins related to loss of cellular polarity. Through this analysis we determined that the viral prevalence in the samples studied was 94.8%. In the characterization of the generated sequences, we analyzed the viral fragments LT3 and VP1 and identified four molecular variants corresponding to three variants and the prototype sequence in each of these. The identified variants were compared to those described in the database and their sequences were used to construct phylogenetic trees. In this way, we verified a wide and random distribution of the variants identified in the branches of these trees. Finally, we analyzed the expression pattern of proteins related to the loss of polarity event. In general, we did not observe significant differences between the marking patterns for the proteins analyzed individually between positive samples in negative for the virus.
 
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Publishing Date
2022-01-13
 
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