INVESTIGATING THE ROLE OF FOXA2 TRANSCRPTION FACTOR IN ADIPOGENESIS USING HUMAN-INDUCED PLURIPOTENT STEM CELLS

Abstract

FOXA2 is a transcription factor that regulates the development and metabolism of various tissues. It has previously been demonstrated to inhibit adipogenesis in preadipocytes. Foxa-2 is also expressed in the adipocytes of mice with high-fat diet-induced obesity, and its haploinsufficiency leads to increased adipogenesis, lipid accumulation, and increased fat mass. However, the role of FOXA2 in human adipocytes has not been studied. Therefore, in this study, we aimed to investigate the role of FOXA2 in adipogenesis using previously established wild-type induced pluripotent stem cells (WT-iPSCs) and FOXA2 knockout iPSCs (FOXA2 KO iPSCs). Our analysis of FOXA2 expression during adipocyte differentiation, using conventional and real-time PCR (RT-PCR), revealed that its highest expression occurs at day 5 during embryoid body (EB) formation, followed by continuous but reduced expression. We noticed that EBs lacking the FOXA2 gene experienced increased cell death. We also found that loss of FOXA2 reduced the proliferation rate of MSCs compared to WT-MSCs, as indicated by the expression of Ki67. Furthermore, mature adipocytes differentiated from FOXA2 KO derived MSCs showed elevated apoptosis indicated by the Annexin V staining, lipid accumulation detected by Oil Red O staining, and decreased expression of adipocyte maturation and functional genes utilizing the RT-PCR technique. These findings provide novel insights into the role of FOXA2 in adipogenesis. We propose that FOXA2 plays a crucial role in adipocyte differentiation by regulating EBs formation and MSCs proliferation. Furthermore, it is essential for maintaining mature adipocyte functionality by modulating key genes involved in adipocyte maturation and metabolism. Further investigations are needed to fully elucidate the molecular mechanisms underlying these effects.

Date of Award	2025
Original language	American English
Awarding Institution	HBKU College of Health & Life Sciences