RGD density along with substrate stiffness regulate hPSC hepatocyte functionality through YAP signalling.

Autor: Blackford SJI; Department of Metabolism, Digestion and Reproduction, Imperial College London, UK; Centre for Craniofacial & Regenerative Biology, King's College London, UK; Centre for Gene Therapy & Regenerative Medicine, King's College London, UK; NIHR Imperial BRC iPSC and Organoid Core Facility, Imperial College London, UK. Electronic address: sji.blackford@outlook.com., Yu TTL; Centre for Craniofacial & Regenerative Biology, King's College London, UK., Norman MDA; Centre for Craniofacial & Regenerative Biology, King's College London, UK., Syanda AM; Department of Metabolism, Digestion and Reproduction, Imperial College London, UK; NIHR Imperial BRC iPSC and Organoid Core Facility, Imperial College London, UK., Manolakakis M; MRC London Institute of Medical Sciences, UK; Institute of Clinical Sciences, Imperial College London, UK., Lachowski D; Cellular and Molecular Biomechanics Laboratory, Department of Bioengineering, Imperial College London, UK., Yan Z; Centre for Craniofacial & Regenerative Biology, King's College London, UK., Guo Y; Centre for Craniofacial & Regenerative Biology, King's College London, UK., Garitta E; Department of Metabolism, Digestion and Reproduction, Imperial College London, UK; NIHR Imperial BRC iPSC and Organoid Core Facility, Imperial College London, UK., Riccio F; Centre for Gene Therapy & Regenerative Medicine, King's College London, UK., Jowett GM; Centre for Craniofacial & Regenerative Biology, King's College London, UK; Centre for Gene Therapy & Regenerative Medicine, King's College London, UK., Ng SS; Department of Metabolism, Digestion and Reproduction, Imperial College London, UK; NIHR Imperial BRC iPSC and Organoid Core Facility, Imperial College London, UK., Vernia S; MRC London Institute of Medical Sciences, UK; Institute of Clinical Sciences, Imperial College London, UK., Del Río Hernández AE; Cellular and Molecular Biomechanics Laboratory, Department of Bioengineering, Imperial College London, UK., Gentleman E; Centre for Craniofacial & Regenerative Biology, King's College London, UK. Electronic address: eileen.gentleman@kcl.ac.uk., Rashid ST; Department of Metabolism, Digestion and Reproduction, Imperial College London, UK; NIHR Imperial BRC iPSC and Organoid Core Facility, Imperial College London, UK. Electronic address: t.rashid@imperial.ac.uk.
Jazyk: angličtina
Zdroj: Biomaterials [Biomaterials] 2023 Feb; Vol. 293, pp. 121982. Date of Electronic Publication: 2022 Dec 22.
DOI: 10.1016/j.biomaterials.2022.121982
Abstrakt: 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.
Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: S.T.R. is a scientific founder, shareholder, and consultant for DefiniGen, Ltd. The other authors indicated no potential conflicts of interest relevant to the study presented here.
(Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
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