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Using Human Induced Pluripotent Stem Cell-derived Hepatocyte-like Cells for Drug Discovery
Published on: May 19, 2018
FOXM1 inhibition primes terminal differentiation of human iPSC-derived hepatocytes
Kayque Alves Telles-Silva1,2, Lara Pacheco1, Sabrina Komatsu1
1Human Genome Research Center (HUG-CEL), Department of Genetics and Evolutionary Biology, Institute of Biosciences, University of São Paulo, São Paulo, SP, Brazil.
Abstract:
Human iPSC-derived hepatocytes are widely used in disease modeling. However, late embryonic development in the human liver remains elusive, which hinders differentiation. During late liver embryonic development, Topoisomerase II (TOP2) is downregulated; however, its role in differentiation is unclear. We replicated the TOP2 silencing at birth and identified a transcription factor crucial for hepatocyte differentiation in vitro. Subtoxic inhibition of TOP2 reduces nuclear chromatin condensation without causing DNA damage. RNA-seq analysis revealed that TOP2 inhibition induced cell cycle arrest, accompanied by FOXM1 downregulation. ATAC-seq confirmed that TOP2A inhibition decreased chromatin accessibility and modulated the Wnt/β-catenin pathway. Proteomic analysis demonstrated that FOXM1 inhibition mimicked TOP2A-mediated cell cycle arrest and reduced the levels of fetal hepatocyte proteins. Prolonged FOXM1 inhibition results in increased hepatocyte polyploidization, enhanced CYP450 activity, and improved lipid metabolism. Our findings suggest that FOXM1 inhibition promotes terminal differentiation of human iPSC-derived hepatocytes, potentially paving the way for understanding late embryonic development in the human liver.
Insights
Inhibiting Topoisomerase II (TOP2) in human iPSC-derived hepatocytes promotes differentiation by downregulating FOXM1. This finding offers insights into late liver development and improves hepatocyte function.
Area of Science:
- Hepatology
- Developmental Biology
- Cell Biology
Background:
- Human induced pluripotent stem cell (iPSC)-derived hepatocytes are vital for disease modeling.
- Late embryonic liver development and differentiation remain poorly understood.
- Topoisomerase II (TOP2) is downregulated during late human liver development, but its role is unclear.
Purpose of the Study:
- To investigate the role of TOP2 in human hepatocyte differentiation.
- To identify key factors regulating late embryonic liver development using iPSC-derived hepatocytes.
Main Methods:
- TOP2 silencing in iPSC-derived hepatocytes.
- RNA-sequencing (RNA-seq) and Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq).
- Proteomic analysis and functional assays (CYP450 activity, lipid metabolism).
Main Results:
- Subtoxic TOP2 inhibition reduced chromatin condensation and induced cell cycle arrest, downregulating FOXM1.
- TOP2 inhibition decreased chromatin accessibility and modulated the Wnt/β-catenin pathway.
- FOXM1 inhibition mimicked TOP2 effects, promoting terminal differentiation, polyploidization, enhanced CYP450 activity, and improved lipid metabolism.
Conclusions:
- TOP2 inhibition promotes terminal differentiation of human iPSC-derived hepatocytes via FOXM1 downregulation.
- This mechanism provides a novel approach to studying late embryonic liver development.
- Targeting FOXM1 may enhance the functionality of iPSC-derived hepatocytes for therapeutic applications.
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