Aberrant development of pancreatic beta cells derived from human iPSCs with FOXA2 deficiency

Abstract

FOXA2 has been identified as an essential factor for pancreas development and emerging evidence supports an association between FOXA2 and diabetes. Although the role of FOXA2 during pancreatic development is well-studied in animal models, its role during human islet cell development remains unclear. Here, we generated induced pluripotent stem cells (iPSCs) from a patient with FOXA2 haploinsufficiency (FOXA2+/− iPSCs) followed by beta-cell differentiation to understand the role of FOXA2 during pancreatic beta-cell development. Our results showed that FOXA2 haploinsufficiency resulted in aberrant expression of genes essential for the differentiation and proper functioning of beta cells. At pancreatic progenitor (PP2) and endocrine progenitor (EPs) stages, transcriptome analysis showed downregulation in genes associated with pancreatic development and diabetes and upregulation in genes associated with nervous system development and WNT signaling pathway. Knockout of FOXA2 in control iPSCs (FOXA2−/− iPSCs) led to severe phenotypes in EPs and beta-cell stages. The expression of NGN3 and its downstream targets at EPs as well as INSUILIN and GLUCAGON at the beta-cell stage, were almost absent in the cells derived from FOXA2−/− iPSCs. These findings indicate that FOXA2 is crucial for human pancreatic endocrine development and its defect may lead to diabetes based on FOXA2 dosage.

Loss of FOXA2 induces ER stress and hepatic steatosis and alters developmental gene expression in human iPSC-derived hepatocytes

“…In contrast to our findings, deletion of Foxa2 in mouse liver does not change the expression levels of liver genes [ 19 ]; however, the expression of developmental genes is significantly altered in mouse liver lacking both Foxa1 and Foxa2 or triple Foxa1, Foxa2, and Foxa3 [ 3 , 20 ], indicating the compensatory role played by the Foxa genes and suggest the difference between mouse and human models. Our results revealed that the loss of FOXA2 was not compensated by FOXA1 and FOXA3, particularly at the MH stage during hepatocyte development as previously reported in human [ 8 ] and rodent models [ 9 , 19 ]. Taken together, our findings indicate that FOXA2 plays an essential role in hepatocyte and biliary development.…”

“…In order to investigate the role of FOXA2 in the development and function of human hepatocytes, we established FOXA2 knockout iPSC models (FOXA2 −/− iPSCs) using CRISPR/Cas9 as we recently reported [ 8 ]. We further confirmed the absence of FOXA2 in HP and MH derived from FOXA2 −/− iPSCs using Western blotting and immunostaining (Fig.…”

“…The advent of induced pluripotent stem cell (iPSC) technology has enabled studying the role of various TFs by recapitulating the process of human development in-vitro. Our lab has recently made use of this technology to demonstrate FOXA2’s role in pancreatic beta cell development, and have found aberrant pancreatic development in the absence of FOXA2 by showing reduced expression of key TFs governing beta cell development [ 8 ]. In the current study, our aim was to use those FOXA2 −/− iPSC models to understand the role of FOXA2 during human hepatocyte development.…”

Toward Precision Medicine with Human Pluripotent Stem Cells for Diabetes

“…Patient-specific hapoloinsufficiency model for FOXA2 showed severe pancreatic phenotypes, with significant reduction in pancreatic progenitors and endocrine lineages. 52 …”

Stem Cell-Derived β Cells: A Versatile Research Platform to Interrogate the Genetic Basis of β Cell Dysfunction

“…Other described iPSC lines were generated from patients with mutations in GCK [ 144 ], PDX1 [ 13 , 155 ], KCNJ11 [ 156 ], INS [ 157 , 158 ], YIPF5 [ 159 ], GATA6 [ 160 ], WFS1 [ 161 , 162 ], TRMT10A [ 163 ], ONECUT [ 136 ], FOXA2 [ 164 ] and activating mutations in STAT3 [ 165 ]. Patient-derived iPSCs were also used to model congenital hyperinsulinism caused by ABCC8 deficiency [ 166 ].…”