Studying the Genetic Defects-Associated with Diabetes in Insulin-Target Cells Using Human Induced Pluripotent Stem Cell Technology

With the continuing surge of evidence on the endocrine and metabolic roles played by adipose and hepatic tissues, it has become increasingly important to understand regulatory pathways governing their development, to find novel therapeutic targets to combat metabolic disorders. Although much progress has been made in understanding their development, findings have been obtained using non-human preadipocyte and hepatocyte models, which make the translation of these findings highly controversial. Therefore, this research project makes use of human induced pluripotent stem cells (iPSCs) towards modelling hepatic and adipocyte associated disorders. Development of hepatocyte is assessed in the absence of FOXA2, an important marker for endodermal development and metabolic gene expression, while development of adipocytes is assessed in the light of multiple heritable gene variants by using induced pluripotent stem cells (iPSCs) derived from the insulin resistant individuals. Our results showed FOXA2 to impact hepatocyte development at progenitor and maturation stages of development by impacting functionality of key transcription factors– HNF16, C/EBPA, HNF1B and functional – ALB, CPS1, TTR markers, whereas next generation sequencing of mature hepatocytes derived from FOXA2+/- iPSC revealed it to impact hepatocyte functionality by impacting genes regulating bile acid homeostasis and gluconeogenesis. Moreover, iPSCs generated from insulin resistant and insulin sensitive individuals gave us fully mature mesenchymal stem cells (MSCs) characterized by high expression of CD90, CD44 and CD73, with low expression of hematopoietic markers. Generated MSCs allowed us to obtain mature adipocytes, whose sorting using Nile red gave a pool of pure adipocytes. This model would allow us to make tangible inferences on the effect of genetic makeup towards adipocyte development, without it getting perturbed by heterogeneity in cell population.