Potential use of iPSC-derived hepatocyte-like cells to develop precision medicine for ASH

Authors

• SH Hoey
• AJ Sanyal
• B Mukhopadhyay
• D Goldman

Abstract

Objectives: Genetic and environmental factors contribute to the development of risky drinking pattern or unhealthy eating behavior, which can result in the development of alcoholic steatohepatitis (ASH) or nonalcoholic steatohepatitis (NASH). We optimized the production of iPSCs from healthy or (N)ASH patient blood samples and their differentiation into hepatocyte-like cells (iHLCs) and probed iPSC-derived iHLCs to understand the interplay of genetic and environmental factors in disease development.
Methods: iPSC clones were generated from CD4+ T cells isolated from the healthy and ASH patient blood samples. iPSCs were sequentially treated with growth factors and activators to generate iHLCs, characterized by the expression of multiple markers as detected by RT-PCR, flow cytometry and confocal microscopy. Functional assays were performed to probe lipid and glycogen accumulation and toxicity. Transcriptome analyses were performed to understand the molecular signature of ASH.
Results: iPSCs were characterized by the expression of SSEA4 and Tra-1-60. iHLCs generated from iPSCs isolated from healthy subjects were characterized by markers albumin and HNF4α. ASH cirrhotic (AC) patients derived hepatocytes are associated with increased lipid droplet (LD) formation, leading to increased senescence. Treatment with Aramchol, a fatty acid pathway modulator, led to decreased LD formation and senescence.
Conclusion: iPSCs generated from whole blood can be induced to differentiate into iHLCs, which can be further characterized by transcriptome analyses. In vitro exposure of iHLCs to ethanol induced lipid accumulation and upregulated fatty acid synthesis pathways. iHLCs generated from ASH patients’ blood samples and their further characterization may facilitate the development of precision therapeutics.

Scientific Focus Area: Stem Cell Biology

This page was last updated on Tuesday, August 6, 2024