A human pluripotent stem cell model for the analysis of metabolic dysfunction in hepatic steatosis

Matthew Sinton; Jose Meseguer-Ripolles; Baltasar Lucendo-Villarin; Sara Wernig-Zorc; John Thomson; Roderick Carter; Macrus Lyall; Paul Walker; Alpesh Thakker; Richard Meehan; Gareth Lavery; Nicholas Morton; Christian Ludwig; Daniel Tennant; David Hay; Amanda Drake

doi:https://doi.org/10.1016/j.isci.2020.101931

A human pluripotent stem cell model for the analysis of metabolic dysfunction in hepatic steatosis

Matthew Sinton, Jose Meseguer-Ripolles, Baltasar Lucendo-Villarin, Sara Wernig-Zorc, John Thomson, Roderick Carter, Macrus Lyall, Paul Walker, Alpesh Thakker, Richard Meehan, Gareth Lavery, Nicholas Morton, Christian Ludwig, Daniel Tennant, David Hay, Amanda Drake

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Abstract

Nonalcoholic fatty liver disease (NAFLD) is currently the most prevalent form of liver disease worldwide. This term encompasses a spectrum of pathologies, from benign hepatic steatosis to non-alcoholic steatohepatitis, which have, to date, been challenging to model in the laboratory setting. Here, we present a human pluripotent stem cell (hPSC)-derived model of hepatic steatosis, which overcomes inherent challenges of current models and provides insights into the metabolic rewiring associated with steatosis. Following induction of macrovesicular steatosis in hepatocyte-like cells using lactate, pyruvate, and octanoate (LPO), respirometry and transcriptomic analyses revealed compromised electron transport chain activity. 13C isotopic tracing studies revealed enhanced TCA cycle anaplerosis, with concomitant development of a compensatory purine nucleotide cycle shunt leading to excess generation of fumarate. This model of hepatic steatosis is reproducible, scalable, and overcomes the challenges of studying mitochondrial metabolism in currently available models.

Original language	English
Article number	101931
Journal	iScience
Volume	24
Issue number	1
Early online date	11 Dec 2020
DOIs	https://doi.org/10.1016/j.isci.2020.101931
Publication status	Published - 22 Jan 2021

Bibliographical note

Funding Information:
MCS was supported by a British Heart Foundation PhD studentship (FS/16/54/32730) and the British Heart Foundation Centre of Research Excellence . SW-Z was funded by the Deutsche Forschungsgemeinschaft ( SFB960 ). This work was supported in part by the Wellcome Trust [grant number 208400/Z/17/Z ], and we thank HWB-NMR at the University of Birmingham for providing open access to their Wellcome Trust-funded NMR equipment. RNC and NMM were supported by a Wellcome Trust New Investigator Award to NMM (100981/Z/13/Z). MJL was supported by a Wellcome Trust PhD Fellowship as part of the Edinburgh Clinical Academic Track scheme (102839/Z/13/Z). RRM is supported by the Medical Research Council and by grants from the BBSRC . Research in RRM's lab leading to these results is partly funded by the Innovative Medicine Initiative Joint Undertaking (IMI JU) under grant agreement number 115001 (MARCAR project: URL: http://www.imi-marcar.eu/ ) . DCH, JMR, and BLV were supported with awards from the MRC Doctoral Training Partnership (MR/K501293/1) and the Chief Scientist Office (TCS/16/37). AJD was funded by the British Heart Foundation Centre of Research Excellence , University of Edinburgh . Our thanks go to the Wellcome Trust Clinical Research Facility Genetics Core, Western General Hospital, Edinburgh, UK. We thank Will Cawthorn for discussions about mitochondrial quantification. Graphical abstract and Figure 1 A were created using the BioRender platform.

Keywords

Endocrine System Physiology
Stem Cells Research
Transcriptomics

ASJC Scopus subject areas

General

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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https://doi.org/10.1016/j.isci.2020.101931Licence: Creative Commons: Attribution-NonCommercial-NoDerivs (CC BY-NC-ND)

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Cite this

Sinton, M., Meseguer-Ripolles, J., Lucendo-Villarin, B., Wernig-Zorc, S., Thomson, J., Carter, R., Lyall, M., Walker, P., Thakker, A., Meehan, R., Lavery, G., Morton, N., Ludwig, C., Tennant, D., Hay, D., & Drake, A. (2021). A human pluripotent stem cell model for the analysis of metabolic dysfunction in hepatic steatosis. iScience, 24(1), Article 101931. Advance online publication. https://doi.org/10.1016/j.isci.2020.101931

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title = "A human pluripotent stem cell model for the analysis of metabolic dysfunction in hepatic steatosis",

abstract = "Nonalcoholic fatty liver disease (NAFLD) is currently the most prevalent form of liver disease worldwide. This term encompasses a spectrum of pathologies, from benign hepatic steatosis to non-alcoholic steatohepatitis, which have, to date, been challenging to model in the laboratory setting. Here, we present a human pluripotent stem cell (hPSC)-derived model of hepatic steatosis, which overcomes inherent challenges of current models and provides insights into the metabolic rewiring associated with steatosis. Following induction of macrovesicular steatosis in hepatocyte-like cells using lactate, pyruvate, and octanoate (LPO), respirometry and transcriptomic analyses revealed compromised electron transport chain activity. 13C isotopic tracing studies revealed enhanced TCA cycle anaplerosis, with concomitant development of a compensatory purine nucleotide cycle shunt leading to excess generation of fumarate. This model of hepatic steatosis is reproducible, scalable, and overcomes the challenges of studying mitochondrial metabolism in currently available models. ",

keywords = "Endocrine System Physiology, Stem Cells Research, Transcriptomics",

author = "Matthew Sinton and Jose Meseguer-Ripolles and Baltasar Lucendo-Villarin and Sara Wernig-Zorc and John Thomson and Roderick Carter and Macrus Lyall and Paul Walker and Alpesh Thakker and Richard Meehan and Gareth Lavery and Nicholas Morton and Christian Ludwig and Daniel Tennant and David Hay and Amanda Drake",

note = "Funding Information: MCS was supported by a British Heart Foundation PhD studentship (FS/16/54/32730) and the British Heart Foundation Centre of Research Excellence . SW-Z was funded by the Deutsche Forschungsgemeinschaft ( SFB960 ). This work was supported in part by the Wellcome Trust [grant number 208400/Z/17/Z ], and we thank HWB-NMR at the University of Birmingham for providing open access to their Wellcome Trust-funded NMR equipment. RNC and NMM were supported by a Wellcome Trust New Investigator Award to NMM (100981/Z/13/Z). MJL was supported by a Wellcome Trust PhD Fellowship as part of the Edinburgh Clinical Academic Track scheme (102839/Z/13/Z). RRM is supported by the Medical Research Council and by grants from the BBSRC . Research in RRM's lab leading to these results is partly funded by the Innovative Medicine Initiative Joint Undertaking (IMI JU) under grant agreement number 115001 (MARCAR project: URL: http://www.imi-marcar.eu/ ) . DCH, JMR, and BLV were supported with awards from the MRC Doctoral Training Partnership (MR/K501293/1) and the Chief Scientist Office (TCS/16/37). AJD was funded by the British Heart Foundation Centre of Research Excellence , University of Edinburgh . Our thanks go to the Wellcome Trust Clinical Research Facility Genetics Core, Western General Hospital, Edinburgh, UK. We thank Will Cawthorn for discussions about mitochondrial quantification. Graphical abstract and Figure 1 A were created using the BioRender platform.",

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Sinton, M, Meseguer-Ripolles, J, Lucendo-Villarin, B, Wernig-Zorc, S, Thomson, J, Carter, R, Lyall, M, Walker, P, Thakker, A, Meehan, R, Lavery, G, Morton, N, Ludwig, C , Tennant, D, Hay, D & Drake, A 2021, 'A human pluripotent stem cell model for the analysis of metabolic dysfunction in hepatic steatosis', iScience, vol. 24, no. 1, 101931. https://doi.org/10.1016/j.isci.2020.101931

TY - JOUR

T1 - A human pluripotent stem cell model for the analysis of metabolic dysfunction in hepatic steatosis

AU - Sinton, Matthew

AU - Meseguer-Ripolles, Jose

AU - Lucendo-Villarin, Baltasar

AU - Wernig-Zorc, Sara

AU - Thomson, John

AU - Carter, Roderick

AU - Lyall, Macrus

AU - Walker, Paul

AU - Thakker, Alpesh

AU - Meehan, Richard

AU - Lavery, Gareth

AU - Morton, Nicholas

AU - Ludwig, Christian

AU - Tennant, Daniel

AU - Hay, David

AU - Drake, Amanda

N1 - Funding Information: MCS was supported by a British Heart Foundation PhD studentship (FS/16/54/32730) and the British Heart Foundation Centre of Research Excellence . SW-Z was funded by the Deutsche Forschungsgemeinschaft ( SFB960 ). This work was supported in part by the Wellcome Trust [grant number 208400/Z/17/Z ], and we thank HWB-NMR at the University of Birmingham for providing open access to their Wellcome Trust-funded NMR equipment. RNC and NMM were supported by a Wellcome Trust New Investigator Award to NMM (100981/Z/13/Z). MJL was supported by a Wellcome Trust PhD Fellowship as part of the Edinburgh Clinical Academic Track scheme (102839/Z/13/Z). RRM is supported by the Medical Research Council and by grants from the BBSRC . Research in RRM's lab leading to these results is partly funded by the Innovative Medicine Initiative Joint Undertaking (IMI JU) under grant agreement number 115001 (MARCAR project: URL: http://www.imi-marcar.eu/ ) . DCH, JMR, and BLV were supported with awards from the MRC Doctoral Training Partnership (MR/K501293/1) and the Chief Scientist Office (TCS/16/37). AJD was funded by the British Heart Foundation Centre of Research Excellence , University of Edinburgh . Our thanks go to the Wellcome Trust Clinical Research Facility Genetics Core, Western General Hospital, Edinburgh, UK. We thank Will Cawthorn for discussions about mitochondrial quantification. Graphical abstract and Figure 1 A were created using the BioRender platform.

PY - 2021/1/22

Y1 - 2021/1/22

N2 - Nonalcoholic fatty liver disease (NAFLD) is currently the most prevalent form of liver disease worldwide. This term encompasses a spectrum of pathologies, from benign hepatic steatosis to non-alcoholic steatohepatitis, which have, to date, been challenging to model in the laboratory setting. Here, we present a human pluripotent stem cell (hPSC)-derived model of hepatic steatosis, which overcomes inherent challenges of current models and provides insights into the metabolic rewiring associated with steatosis. Following induction of macrovesicular steatosis in hepatocyte-like cells using lactate, pyruvate, and octanoate (LPO), respirometry and transcriptomic analyses revealed compromised electron transport chain activity. 13C isotopic tracing studies revealed enhanced TCA cycle anaplerosis, with concomitant development of a compensatory purine nucleotide cycle shunt leading to excess generation of fumarate. This model of hepatic steatosis is reproducible, scalable, and overcomes the challenges of studying mitochondrial metabolism in currently available models.

AB - Nonalcoholic fatty liver disease (NAFLD) is currently the most prevalent form of liver disease worldwide. This term encompasses a spectrum of pathologies, from benign hepatic steatosis to non-alcoholic steatohepatitis, which have, to date, been challenging to model in the laboratory setting. Here, we present a human pluripotent stem cell (hPSC)-derived model of hepatic steatosis, which overcomes inherent challenges of current models and provides insights into the metabolic rewiring associated with steatosis. Following induction of macrovesicular steatosis in hepatocyte-like cells using lactate, pyruvate, and octanoate (LPO), respirometry and transcriptomic analyses revealed compromised electron transport chain activity. 13C isotopic tracing studies revealed enhanced TCA cycle anaplerosis, with concomitant development of a compensatory purine nucleotide cycle shunt leading to excess generation of fumarate. This model of hepatic steatosis is reproducible, scalable, and overcomes the challenges of studying mitochondrial metabolism in currently available models.

KW - Endocrine System Physiology

KW - Stem Cells Research

KW - Transcriptomics

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DO - https://doi.org/10.1016/j.isci.2020.101931

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SN - 2589-0042

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ER -

A human pluripotent stem cell model for the analysis of metabolic dysfunction in hepatic steatosis

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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Cite this