Functional study in penis carcimomas for the validation of candidate molecular markers obtained from integrated global analyzes (genomics, transcriptomics and DNA methylation)

Abstract

Penile carcinomas (PeCa) is a rare disease in developed countries and highly frequent in poor and developing countries, including Brazil. Despite of its unpredictable behavior and aggressive treatment, there is a very limited number of data on the genetic and epigenetic mechanisms involved this disease. Previosuly, we used large-scale studies (methylation profile, expression, miRNA and genomic gains and losses) in CaPe aiming to identify molecular markers involved in the development and progression of this disease. Algorithms and bioinformatics strategies were applied to integrate the genomic, transcriptomic and methylation data obtained in these studies in order to identify potential drivers in CaPe. Here, we will validate the drivers obtained from the integrative analysis. In addition, we will establish cell lines using xenotransplants strategy. In the present study, we will apply RT-qPCR (mRNA and miRNA), pyrosequencing (methylation), and methodologies for isolation of cancer stem cells (flow cytometry), cultivation and expansion of CaPe cells in vivo using immunodeficient mice. Tumors generated in animals will be evaluated by large-scale techniques aiming to verify if its recapitulate the original tumor. The use of large scale methodologies is the main stage for the discovery of markers that can contribute to the understanding of the genetic and epigenetic mechanisms involved in CaPe. The results from these methodologies, coupled with the use of animal models will provide subsidies for the design of specific targeted therapies and functional analysis, that are until this moment, undescribed in this tumor.