INDUCED PLURIPOTENT STEM CELL DERIVED LIVER MODEL FOR THE STUDY OF PNPLA3-ASSOCIATED NON-ALCOHOLIC FATTY LIVER DISEASE
type: CreativeWork
description: Non-alcoholic fatty liver disease (NAFLD) is now the leading cause of chronic liver disease in the developed world, afflicting approximately one in four adults globally. NAFLD is defined by the accumulation of fat within the liver which ranges in severity from simple steatosis to non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma. Until recently, NAFLD has been considered to be a consequence of metabolic syndrome; however, recent studies suggest that genetic factors may also influence disease onset and progression. Accordingly, genome wide association studies have linked the I148M variant in the gene coding for Patatin-like phospholipase domain-containing protein 3 (PNPLA3) with NAFLD aggravation without underlying metabolic disease. However, despite over a decade of research, the function of PNPLA3 and its role in the pathogenesis of NAFLD remains largely obscure. The lack of clarity regarding the function and disease associations of PNPLA3 is due in large part to the lack of a comprehensive human model of PNPLA3-associated NAFLD. In order to address this need, we have developed an in vitro model that takes advantage of the unique properties of human induced pluripotent stem cells (hiPSC) and the CRISPR/CAS9 gene editing technology. We first used CRISPR/CAS9 to generate hiPSC lines with either a knock-out (PNPLA3KO) of the PNPLA3 gene or with the I148M variant knocked in (PNPLA3I148M). The resulting cells were then differentiated into hepatocytes and grown in 3D culture conditions to improve their maturity and functionality. The differentiated cells were treated with either monounsaturated or saturated free fatty acids to induce NAFLD-like phenotypes (lipid accumulation and lipid toxicity, respectively) and characterized by various functional, genomic, and lipidomic assays. The genetically edited sublines showed similar differentiation efficiency toward hepatocytes as untargeted cells indicating that changes in PNPLA3 activity does not af…
abstract: Non-alcoholic fatty liver disease (NAFLD) is now the leading cause of chronic liver disease in the developed world, afflicting approximately one in four adults globally. NAFLD is defined by the accumulation of fat within the liver which ranges in severity from simple steatosis to non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma. Until recently, NAFLD has been considered to be a consequence of metabolic syndrome; however, recent studies suggest that genetic factors may also influence disease onset and progression. Accordingly, genome wide association studies have linked the I148M variant in the gene coding for Patatin-like phospholipase domain-containing protein 3 (PNPLA3) with NAFLD aggravation without underlying metabolic disease. However, despite over a decade of research, the function of PNPLA3 and its role in the pathogenesis of NAFLD remains largely obscure. The lack of clarity regarding the function and disease associations of PNPLA3 is due in large part to the lack of a comprehensive human model of PNPLA3-associated NAFLD. In order to address this need, we have developed an in vitro model that takes advantage of the unique properties of human induced pluripotent stem cells (hiPSC) and the CRISPR/CAS9 gene editing technology. We first used CRISPR/CAS9 to generate hiPSC lines with either a knock-out (PNPLA3KO) of the PNPLA3 gene or with the I148M variant knocked in (PNPLA3I148M). The resulting cells were then differentiated into hepatocytes and grown in 3D culture conditions to improve their maturity and functionality. The differentiated cells were treated with either monounsaturated or saturated free fatty acids to induce NAFLD-like phenotypes (lipid accumulation and lipid toxicity, respectively) and characterized by various functional, genomic, and lipidomic assays. The genetically edited sublines showed similar differentiation efficiency toward hepatocytes as untargeted cells indicating that changes in PNPLA3 activity does not af…
author: Tilson, Samantha Grace
language: eng
keywords: iPSC, NAFLD, PNPLA3, in vitro disease modelling, hepatology
license: https://www.repository.cam.ac.uk/bitstreams/e534359b-423b-4931-94f2-758319ea64da/download
license: https://www.rioxx.net/licenses/all-rights-reserved/
date published: 2020-08-31
date modified: 2026-07-24T01:33:13Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Cambridge

PropertyValue
property ID: DOI
value: 10.17863/CAM.63096