Functional characterization of miR-184 in the prostate of rats subjected to maternal malnutrition. Potential participation in the developmental origin of prostate cancer

Abstract

The DOHaD approach (Developmental Origins of Health and Disease) seeks to establish the correlation between the incidence of diseases in adult life and in aging in view of the events suffered during early development. One of his most used models for DOHaD studies is the submission of rodents to maternal protein restriction (PMR). Experimental studies have already shown that it is responsible for inhibiting prostate development and increasing the incidence of Prostate Cancer (PCa) with aging. One of the main consequences of these DOHaD events is the epigenetic changes that accompany the offspring throughout life, such as microRNAs. In recent years, our research group has been dedicated to comprehensively understanding the molecular and epigenetic mechanisms altered in the prostate of animals subjected to RPM. Thus, the objective of this work will be to analyze the functional role of miR-184 (previously selected from bioinformatics analyses) in the ventral prostate (PV) of animals on a postnatal day (PND) 21 that were submitted to RPM. For this, we will use data from the sequencing of miRNAs (GSE180673) and mRNAs (GSE180674) from the PV of Sprague Dawley rats that 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% protein diet, n=6), which were euthanized at DPN21. These data are part of our research group's database (generated in the FAPESP process 2017/01063-7). From small RNA sequence data, we identified differentially expressed (DE) miRNAs, performed target prediction, compared to PV mRNA sequencing data from these animals, and selected miR-184 to be validated and functionally characterized in prostate cells. Validations will take place through RT-qPCR in the PV of animals in DPN21. In addition, we will also perform functional assays in benign and tumoral prostatic cells (PNT2, LNCaP, and PC3) treated with miR-184 mimetics, where miRNA expression and its predicted targets will be evaluated, as well as cell viability and migration assays. With this work, we hope to characterize the role of miR-184 in the PV of animals submitted to RPM and its functional role in prostatic cells.