Targeting CDX2 in human pluripotent stem cell-derived hepatocyte-like cells

The differentiation of functionally mature hepatocytes from human induced pluripotent stem cells (hiPSC) has the potential to replace primary human hepatocytes (PHH) as the gold standard in vitro model for drug screening, studies of hepatotoxicity as well as liver disease, and is considered a gateway technology to future cell therapy applications. We recently reported that current protocols for deriving hepatocyte-like cells (HLC) from hiPSC induce hybrid differentiation, which leads to the establishment of hepatic and intestinal gene expression signatures within individual HLC, compromising functional maturity. Here, we show that intestinal hybrid differentiation in HLC is under control of caudal-domain type 2 (CDX2). We demonstrate that knock-out of CDX2 in hiPSC effectively disrupts intestinal gene expression profiles, upregulates transcription factors important for hepatic lineage specification in HLC, and leads to the formation of bile canaliculus-like structures in HLC that closer resemble those formed by PHH, including size reduction, loss of intestine-associated membrane transporters and increase in hepatic transporters. The dataset comprises RNA sequencing results from four biological replicates of wild-type and CDX2(-/-) iPSC, four biological replicates of wild-type and CDX2(-/-) HLC complemented with four independent PHH donors.