Integrative analysis of the transcriptome and proteome of pancreatic islets isolated from the offspring of male rats subjected to maternal protein restriction: characterization and functional studies

• Grant number: 25/00542-5
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: June 01, 2025
• End date: March 31, 2029
• Field of knowledge: Biological Sciences - Morphology - Histology
• Principal Investigator: Luis Antonio Justulin Junior
• Grantee: Ana Beathriz Leite Lorente
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

The concept of DOHaD (Developmental Origins of Health and Disease) aims to understand how early developmental events are associated with the occurrence of diseases in adulthood and aging. In this context, maternal protein restriction (MPR) in rodents is widely used as an experimental model to investigate these processes. Evidence indicates that MPR negatively affects insulin secretion and increases the vulnerability of pancreatic beta cells to oxidative stress, contributing to the onset of metabolic syndromes later in aging, such as type 2 diabetes (T2DM). This study aims to analyze the messenger RNA expression profile in pancreatic islets of rats subjected to MPR, followed by integration with proteomic data to identify molecular pathways associated with pancreatic dysfunctions. For this, Sprague Dawley rats will be used, divided into Control (17% protein) and MPR (6% protein) groups during gestation and lactation. On postnatal day 21, the offspring will be weighed, euthanized, and the pancreas collected and processed for morphological and molecular analyses. Pancreatic islets will be isolated for RNA extraction, library preparation, and sequencing (HiSeq2500). Differentially expressed genes will be identified (DESeq2, p < 0.05), and transcriptomic data will be integrated with the proteome (FAPESP process: 2023/06412-0). Based on the integration of omics data, molecular targets will be selected for functional analyses using an in vitro model with immortalized pancreatic beta cells (RIN-5F) and human cells, in collaboration with the Joslin Diabetes Center (USA), in a BEPE project to be submitted. The findings will be validated by RT-qPCR and immunofluorescence. This study seeks to understand the molecular mechanisms underlying intrauterine programming and its possible role in the dysregulation of pancreatic biomarkers related to the development of non-communicable chronic diseases, according to the DOHaD concept. These results may contribute to understanding the importance of nutritional care during pregnancy and early childhood, as well as reinforcing the relevance of public policies for preventing metabolic diseases from the early stages of development.