Hay, David

Hughes, Jeremy

Meseguer Ripollés, Jose

2020-02-10T12:11:42Z

2020-02-10T12:11:42Z

2020-01-22

Liver disease represents a major cause of mortality and morbidity. Despite the regenerative capacity of the liver, maintained injury or acute injury can lead to loss of liver function and disease. The most common cause of acute liver damage is drug-induced liver injury (DILI). This can lead to organ failure and possible death. Therefore, new therapies to reduce the severity of the injury are required. Stimulation of anti-inflammatory and anti-oxidative stress pathways during the resolution of the injury have been proposed as powerful approaches to reduce organ injury and to enhance regeneration. A main transcription factor which regulates anti-inflammatory and anti-oxidative stress is ‘nuclear factor erythroid-derived 2-like 2’ (Nrf2). Therefore, pharmacological activation of the Nrf2 pathway offers the potential to exert a cytoprotective effect promoting tissue regeneration. Dimethyl fumarate (DMF) is a drug approved for some forms of multiple sclerosis. DMF’s protection is due in part by activation of the Nrf2 pathway. We hypothesize that DMF could be used to reduce the severity of DILI via Nrf2 activation. This thesis explores the protective effects of DMF and Nrf2 signalling during paracetamol-induced hepatotoxicity using in vitro and in vivo models. For the in vitro studies, a semi-automated platform to produce hepatocytes-like cells (HLCs) from human pluripotent stem cells was employed. Single-cell high content image analysis was performed to understand Nrf2 nuclear translocation dynamics following DMF administration. The protective properties of DMF were tested in three different combinations: pre-treatment prior to paracetamol incubation, co-treatment or post-treatment following paracetamol injury. In all cases, DMF protected HLCs from paracetamol exposure. These findings were validated in a Zebrafish model of paracetamol injury. A zebrafish liver GFP reporter line was employed to detect fluorescence changes upon paracetamol exposure. Pre-treatment with DMF prior to paracetamol injury reduced the level of GFP loss. RNA sequencing from both models identified that DMF protection was mediated via Nrf2 pathway stimulation. This was mainly by an increase in cell metabolism and oxidative stress management as well as reducing pro-inflammatory pathways activation. In summary, the findings of this work provide new understanding on the effects of DMF in the modulation of the Nrf2 pathway during paracetamol-induced liver injury. These studies may provide a platform to develop new treatment regimes for patients with acute liver disease.

https://hdl.handle.net/1842/36765

http://dx.doi.org/10.7488/era/70

en

The University of Edinburgh

Semi-automated Production of Hepatocyte Like Cells from Pluripotent Stem Cells. Jose Meseguer-Ripolles, Baltasar Lucendo- Villarin, Yu Wang, David C. Hay. J. Vis. Exp. (137), e57995, doi:10.3791/57995 (2018).

Pluripotent Stem Cell-Derived Human Tissue: Platforms to Evaluate Drug Metabolism and Safety. Jose Meseguer-Ripolles, Salman R. Khetani, Javier G. Blanco, Miari Iredale, David C. Hay. Review, AAPS J (2018) 20: 20. https://doi.org/10.1208/s12248-017- 0171-8

Modelling non-alcoholic fatty liver disease in human hepatocytelike cells. Lyall Marcus J., Cartier Jessy, Thomson John P., Cameron Kate, Meseguer-Ripolles Jose, O'Duibhir Eoghan, Szkolnicka Dagmara, Villarin Baltasar Lucendo, Wang Yu, Blanco Giovanny Rodriguez, Dunn Warwick B., Meehan Richard R., Hay David C. ,and Drake Amanda J. Philosophical Transactions B DOI: 10.1098/rstb.2017.0362

A human iPSC line capable of differentiating into functional macrophages expressing ZsGreen: a tool to study and track therapeutic cells in vivo. Martha Lopez Yrigoyen*, Antonella Fidanza*, Luca Cassetta, Richard A. Axton, A. Helen Taylor, Jose Meseguer- Ripolles, Anestis Tsakiridis, Val Wilson, David Hay, Jeff W. Pollard, Lesley M. Forrester. Philosophical Transactions B DOI: 10.1098/rstb.2017-02195

Modelling foetal exposure to maternal smoking using hepatoblasts from pluripotent stem cells. Baltasar Lucendo-Villarin, Panagiotis Filis, Madeleine J. Swortwood, Marilyn A. Huestis, Jose Meseguer-Ripolles, Kate Cameron, John P. Iredale, Peter J. O’Shaughnessy, Paul A. Fowler, David C. Hay. Arch Toxicol (2017) 91: 3633. https://doi.org/10.1007/s00204-017-1983-0

Defined and Scalable Generation of Hepatocyte-like Cells from Human Pluripotent Stem Cells, Yu Wang, Sharmin Alhaque, Kate Cameron, Jose Meseguer-Ripolles, Baltasar Lucendo-Villarin, Hassan Rashidi, David C. Hay. J. Vis. Exp. (121), e55355, doi:10.3791/55355 (2017).

Serum Free Production of Three-Dimensional Human Hepatospheres from Pluripotent Stem Cells. Balta Lucendo-Villarin, Hassan Rashidi, Sharmin Alhaque, Lena Fischer, Jose Meseguer- Ripolles, Yu Wang, Cliona O'Farrelly, Michael Themis, David C. Hay. J. Vis. Exp. (2019)

Genome editing in pluripotent stem cells provides new understandding of protein SUMOylation during stem cell specification. Yu Wang, Michael H. Tatham, Wolfgang Schmidt-Heck, Carolyn Swann, Karamjit Singh-Dolt, Jose Meseguer-Ripolles, Baltasar Lucendo-Villarin, Tilo Kunath, Timothy R. Rudd, Andrew Smith, Jan G. Hengstler, Patricio Godoy, Ronald T. Hay, David C. Hay. iScience, 2019.

stem cells

hepatocyte

Nrf2

liver

dimethyl fumarate

Investigating the protective properties of dimethyl fumarate and Nrf2 signalling in response to drug toxicity

Thesis or Dissertation

Doctoral

PhD Doctor of Philosophy