Metabolic programming and metabolism disruption in adult life: the role of miRNA 483 on the proteomic profile of adipose tissue

Abstract

Numerous studies have shown that disrupted metabolic environments during gestation and lactation that influence growth of the fetus and neonate increase the risk of developing metabolic diseases in adulthood. The mechanisms underlying this relationship are not fully understood. However programmed changes in epigenetic factors, such as miRNAs are thought to play an important role. Previous studies, using a rat model of maternal protein restriction, showed that nutritionally-induced low birth weight led to a programmed increase in the micro RNA miR-483 in adipose tissue. This was associated with a reduction in the capacity of adipose tissue to store lipid. A similar increase in miR-483 was present in adipose tissue from low birth weight humans. This project aims to identify the spectrum of proteins whose translation is regulated by miR-483. This will be achieved using a recently generated transgenic mouse that over-expresses miR-483 in adipose tissue. The state of the art technology, pSILAC will be used to identify miRNA targets by comparing cultured pre-adipocytes from epididymal fat from animals over-expressing miR-483 in adipose tissue mice and wild-type controls. Following data generation from the pSILAC protocol, bioinformatic screening will be performed in order to identify potential direct targets of the miRNA as well as potential dysregulated pathways. Target will be validated by western blotting and luciferase assays will be performed in order to confirm the direct regulation of the target translation by the miRNA. These studies will therefore provide insight into the mechanisms by which dysregulation of miR-483 can lead to programmed changes in adipocyte function.