The role of estrogen in the modulation of miRNAs in the prostate of rats offspring submitted to RPM: evaluation and functional studies in prostate cells

Abstract

The DOHaD (Developmental Origin of Health and Disease) approach seeks to establish the correlation between the incidence of diseases in adult life and in aging facing the events suffered during the beginning of development. One of its most used models for DOHaD studies is the rodent submission maternal protein restriction (PMR). Experimental studies have already shown that PMR is responsible for inhibiting prostate development and increasing the incidence of Prostate Cancer (PCa) with aging. One of the main consequences of these events are the epigenetic changes that accompany the offspring throughout life, such as non-coding RNAs and microRNAs. Recently, our research group is studying in a global way the molecular and epigenetic mechanisms of alterations in the prostate of animals submitted to PMR. Thus, the objective of this work will be to analyze the role of estrogen in the modulation of expression of miR33-5p and miR-184 (previously selected from bioinformatics analyses) in ventral prostate (PV) of animals on postnatal day (PND) 21 that underwent RPM. For that, we will use sequencing data of miRNAs (GSE180673) and mRNAs (GSE180674) from PV of Sprague Dawley rats, who were divided into two experimental groups: CTR group (control, with a 17% protein diet, n=6) and GLLP (protein restriction during pregnancy and lactation, 6% diet, n=6), which were euthanized in the PND 21. These data are part of our research group's database (generated in the FAPESP process 2017/01063-7). From the miRNA sequencing data we identified the targets differentially expressed (DE), we performed target prediction, compared to PV mRNA sequencing data of these animals and selected miRNAs to be validated. Validations will take place through RTq-PCR in the PV of the animals. In addition, we will also perform functional assays in prostate cells tumors (PNT2 and PC3) chronically exposed to estrogen, as well as feasibility and cell migration. With this work, we hope to identify the role of in the modulation of miRNAs in the VP of animals submitted to RPM and its functional role for prostate cells.