Integrative analysis of the transcriptome and proteomics of the ventral prostate of rat offspring submitted to maternal protein restriction

Abstract

Maternal exposure to protein-deficient diet during pregnancy, which is characteristic of underdeveloped countries and countries whose diet contains high-calorie, fiber-poor and nutrient foods, is associated with the development of diseases in the offspring, especially cardiovascular, renal, Diabetes and even Cancer in the offspring. In addition, studies in our group demonstrate that maternal protein restriction affects development and increases the incidence of prostatic lesions in the offspring rats during aging. Despite this, the molecular mechanisms that trigger these changes are still poorly understood. In this context, the advances in methodologies for the study of gene expression and protein on a large scale, combined with the possibility of integrating these data using bioinformatics tools, has enabled a global integrative view of molecular mechanisms under normal and pathological conditions. Thus, the objective of this work will be to identify the global expression profile of messenger RNAs (transcriptome) and proteins (proteomics) in rat prostate samples that undergo perinatal protein restriction and integrate the global expression profile of mRNAs and proteins to identify networks regulatory pathways and molecular pathways involved in prostate development in normal and restricted animals. Male Sprague Dawley rats with 21 days of postnatal age born from mothers fed standard chow (17% protein) or from mothers fed lowprotein chow (6% protein) during gestation and lactation will be analyzed. After this period, the animals will be euthanized with anesthetic overdose and the ventral prostate collected. The transcriptome will be analyzed by next generation sequencing (HigSeq-2500, Ilumina) and proteome by mass spectrometry (LC-Ms/Ms). Some targets differentially expressed will be selected to be validated by RT-qPCR, as well as immunohistochemistry and western blotting. These results will be compared to data from the transcriptome and proteome for prostatic adenocarcinoma available in the literature to investigate whether perinatal protein restriction is responsible for altering the gene and protein expression of targets related to the development of prostatic lesions. Other animals will be euthanized at 540 days of age in order to identify possible altered targets both by protein restriction in young rats and in prostate cancer. This PhD project is part of a regular project (2017/01063-7), which will also analyze the microRNome and the global DNA methylation profile in the prostate of rats submitted to maternal protein restriction. We expected to elucidate the global effects of protein restriction on molecular pathways involved in the prostate development and the repercussions with aging. (AU)