Global methylation and microRNAs expression in ventral prostate of rats offspring submitted to maternal protein restriction: perinatal effects and reflexes on aging

Abstract

In the last decades, it has been observed an increase in the incidence of diseases related to changes in metabolism, mainly in children. Among the most prevalent are Obesity, Diabetes, cardiovascular disease and even some types of Cancer. Epidemiological evidence shows that these chronic diseases can originate from insults suffered by individuals during the intrauterine period, a condition known as Fetal Programming (FP). This period is characterized by the great plasticity and ability of the embryo/foetus to adapt to environmental changes (diet, stress and hormones) by altering gene expression by epigenetic mechanisms. Recent studies demonstrated changes in methylation of promoter genes and microRNAs expression associated with DNA repair, cell cycle and amino acid metabolism in the liver of rats programmed by intrauterine protein restriction. Considering the previous results from our group that demonstrated the role of maternal protein restriction in altering the development and increasing the incidence of prostatic lesions in the aged offspring rats, the objective of this project will be to identify the global methylation and expression profile of microRNAs in prostate samples of offspring rat that underwent maternal protein restriction. Male Sprague Dawley rats with 21 days old born from mothers fed standard chow (17% protein) or with lowproteic chow (6% protein) during gestation and lactation will be used. After weaning, the animals will be euthanized with anaesthetic overdose and the ventral prostate will be collected. The global methylation will be addressed by ELISA (Cell Biolabs - STA380), and the global expression of microRNAs (MIcroRNome) will be determined by next generation sequencing (HigSeq-2500 Illumina). In silico analyses will be performed for the prediction of targets regulated by microRNA, and by methylation. In addition, some targets of interest will be selected to be validated by RT-qPCR, as well as protein expression by immunohitochemistry and western blotting. We will also investigate whether these changes persist with aging in rats at 540 days old. This PhD project is part of a regular project (FAPESP 2017/01063-7), which will also analyse the transcriptome and proteome in the prostate of these offspring rats submitted to perinatal 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)