RGD density along with substrate stiffness regulate hPSC hepatocyte functionality through YAP signalling
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Published version
Author(s)
Type
Journal Article
Abstract
Human pluripotent stem cell-derived hepatocytes (hPSC-Heps) may be suitable for treating liver diseases, but differentiation protocols often fail to yield adult-like cells. We hypothesised that replicating healthy liver niche biochemical and biophysical cues would produce hepatocytes with desired metabolic functionality. Using 2D synthetic hydrogels which independently control mechanical properties and biochemical cues, we found that culturing hPSC-Heps on surfaces matching the stiffness of fibrotic liver tissue upregulated expression of genes for RGD-binding integrins, and increased expression of YAP/TAZ and their transcriptional targets. Alternatively, culture on soft, healthy liver-like substrates drove increases in cytochrome p450 activity and ureagenesis. Knockdown of ITGB1 or reducing RGD-motif-containing peptide concentration in stiff hydrogels reduced YAP activity and improved metabolic functionality; however, on soft substrates, reducing RGD concentration had the opposite effect. Furthermore, targeting YAP activity with verteporfin or forskolin increased cytochrome p450 activity, with forskolin dramatically enhancing urea synthesis. hPSC-Heps could also be successfully encapsulated within RGD peptide-containing hydrogels without negatively impacting hepatic functionality, and compared to 2D cultures, 3D cultured hPSC-Heps secreted significantly less fetal liver-associated alpha-fetoprotein, suggesting furthered differentiation. Our platform overcomes technical hurdles in replicating the liver niche, and allowed us to identify a role for YAP/TAZ-mediated mechanosensing in hPSC-Hep differentiation.
Date Issued
2023-02
Date Acceptance
2022-12-22
Citation
Biomaterials, 2023, 293, pp.1-17
ISSN
0142-9612
Publisher
Elsevier
Start Page
1
End Page
17
Journal / Book Title
Biomaterials
Volume
293
Copyright Statement
0142-9612/© 2022 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/36640555
PII: S0142-9612(22)00622-6
Subjects
Hepatocyte
Hydrogel
Induced pluripotent stem cell
Mechano sensing
YAP
Humans
Colforsin
Hepatocytes
Cell Differentiation
Oligopeptides
Cytochrome P-450 Enzyme System
Hydrogels
Publication Status
Published
Coverage Spatial
Netherlands
Article Number
121982
Date Publish Online
2022-12-22