An in vitro model of hepatic steatosis using lipid loaded induced pluripotent stem cell derived hepatocyte like cells
Main Article Content
Keywords
iPSC, hepatocyte-like cells, hepatic steatosis
Abstract
Hepatic steatosis is a metabolic disease, characterized by selective and progressive accumulation of lipids in liver, leading to progressive non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and cirrhosis. The existing in vitro models of hepatic steatosis to elucidate the molecular mechanisms behind the onset of hepatic steatosis and to profile small molecule modulators uses lipid loaded primary hepatocytes, and cell lines like HepG2. The limitation of these models includes high variability between the different donor samples, reproducibility, and translatability to physiological context. An in vitro human hepatocyte derived model that mimics the pathophysiological changes seen in hepatic steatosis may provide an alternative tool for pre-clinical drug discovery research. We report the development of an in vitro experimental model of hepatic steatosis using human induced pluripotent stem cell (iPSC) derived hepatocytes like cells (HLC), loaded with lipids. Our data suggests that HLC carry some of the functional characteristics of primary hepatocytes and are amenable for development of an in vitro steatosis model using lipid loading method. The in vitro experimental model of hepatic steatosis was further characterized using biomarker analysis and validated using telmisartan. With some refinement and additional validation, our in vitro steatosis model system may be useful for profiling small molecule inhibitors and studying the mechanism of action of new drugs.
Downloads
Metrics
References
2 Portillo-Sanchez P, Cusi K. (2016) Treatment of Nonalcoholic Fatty Liver Disease (NAFLD) in patients with Type 2 Diabetes Mellitus. Clin Diabetes Endocrinol., 12; 2:9
3 Kanuri G, Bergheim I. (2013) In Vitro and in Vivo Models of Non-Alcoholic Fatty Liver Disease (NAFLD). Int. J. Mol. Sci., 14, 11963-11980.
4 Chavez-Tapia NC, Rosso N, Tiribelli C. (2011) In Vitro Models for the Study of Non-Alcoholic Fatty Liver Disease. Current Medicinal Chemistry, 18, 1079-1084
5 Donato MT1, Jover R, Gómez-Lechón MJ. (2013) Hepatic cell lines for drug hepatotoxicity testing: limitations and strategies to upgrade their metabolic competence by gene engineering. Curr Drug Metab., 14(9):946-68
6 Aninat C, Piton A, Glaise D, Le Charpentier T, Langouet S, Morel F, Guguen-Guillouzo C, Guillouzo A. (2006) Expression of cytochromes P450, conjugating enzymes and nuclear receptors in human hepatoma HepaRG cells. Drug Metab Dispos 34:75–83. doi: 10.1124/dmd.105.006759
7 Anthérieu S., Rogue A., Fromenty B., Guillouzo A., Robin M.A. (2011) Induction of vesicular steatosis by amiodarone and tetracycline is associated with up-regulation of lipogenic genes in heparg cells. Hepatology 53:1895–1905. doi: 10.1002/hep.24290
8 Rodrigues RM, Branson S, De Boe V, Sachinidis A, Rogiers V, De Kock J, Vanhaecke T. (2015) In vitro assessment of drug-induced liver steatosis based on human dermal stem cell-derived hepatic cells. Arch Toxicol
9 H. H. J. Gerets, K. Tilmant, B. Gerin, et al., (2012) Characterization of primary human hepatocytes, HepG2 cells, and HepaRG cells at the mRNA level and CYP activity in response to inducers and their predictivity for the detection of human hepatotoxins. Cell Biol Toxicol., 28(2): 69–87
10 Donato MT, Jiménez N, Serralta A, Mir J, Castell JV, Gómez-Lechón MJ. (2007) Effects of steatosis on drug-metabolizing capability of primary human hepatocytes. Toxicol In Vitro 21(2):271-6
11 Marc C. Hansel, Roberto Gramignoli, Kristen J. Skvorak et al. (2014) The History and Use of Human Hepatocytes for the Study and Treatment of Liver Metabolic Diseases. Curr Protoc Toxicol. 62: 14.12.1–14.12.2
12 Sauer V, Roy-Chowdhury N, Guha C, Roy-Chowdhury J., Induced pluripotent stem cells as a source of hepatocytes. Curr Pathobiol Rep., 2(1): 11–20, 2014
13 Chaudhari P, Prasad N, Tian L, Jang YY. (2016) Determination of Functional Activity of Human iPSC-Derived Hepatocytes by Measurement of CYP Metabolism. Methods Mol Biol., 1357:383-94
14 Sirenko O, Cromwell EF. (2018) Determination of Hepatotoxicity in iPSC-Derived Hepatocytes by Multiplexed High Content Assays. Methods Mol Biol., 1683:339-354
15 Sakurai F, Mitani S, Yamamoto T et al., (2017) Human induced-pluripotent stem cell-derived hepatocyte-like cells as an in vitro model of human hepatitis B virus infection. Sci Rep., 7: 45698
16 Nagamoto Y, Takayama K, Ohashi K, et al. (2016) Transplantation of a human iPSC-derived hepatocyte sheet increases survival in mice with acute liver failure. J Hepatol., 64(5):1068-1075
17 Kiamehr M, Viiri LE, Vihervaara T et al. (2017) Lipidomic profiling of patient-specific iPSC-derived hepatocyte-like cells. Dis Model Mech 1;10(9):1141-1153
18 Max A. Cayo, Sunil K. Mallanna, Francesca Di Furio et al. (2017) A Drug Screen using Human iPSC-Derived Hepatocyte-like Cells Reveals Cardiac Glycosides as a Potential Treatment for Hypercholesterolemia. Cell Stem Cell., 6;20(4):478-489.e5
19 Liu JT, Lamprecht MP, Duncan SA. (2018) Using Human Induced Pluripotent Stem Cell-derived Hepatocyte-like Cells for Drug Discovery. J Vis Exp. 19;(135)
20 Alam S, Kabir J, Mustafa G, Gupta U, Hasan SK, Alam AK. (2016) Effect of Telmisartan on histological activity and fibrosis of non-alcoholic steatohepatitis: A 1-year randomized control trial. Saudi J Gastroenterol 22(1):69-76
21 Zhong S, Han W, Hou C et al., (2017) Relation of Transcriptional Factors to the Expression and Activity of Cytochrome P450 and UDP-Glucuronosyltransferases 1A in Human Liver: Co-Expression Network Analysis. AAPS J. Jan;19(1):203-214
22 Daiki Nakamori, Hiroki Akamine, Kazuo Takayama et al. (2017) Direct conversion of human fibroblasts into hepatocyte-like cells by ATF5, PROX1, FOXA2, FOXA3, and HNF4A transduction. Scientific Reports volume 7, Article number: 16675
23 Kubo A, Kim YH, Irion S et al. (2010) The homeobox gene Hex regulates hepatocyte differentiation from embryonic stem cell-derived endoderm. Hepatology. Feb;51(2):633-41
24 Ishikawa T, Kobayashi M, Yanagi S et al. (2015) Human induced hepatic lineage-oriented stem cells: autonomous specification of human iPS cells toward hepatocyte-like cells without any exogenous differentiation factors. PLoS One. Apr 13;10(4):e0123193
25 Chiba M, Ishii Y, Sugiyama Y. (2009) Prediction of hepatic clearance in human from in vitro data for successful drug development. AAPS J. Jun;11(2):262-76
26 Kurogi K, Chen M, Lee Y, et al., Sulfation of buprenorphine, pentazocine, and naloxone by human cytosolic sulfotransferases. Drug Metab Lett. 2012 Jun 1;6(2):109-15
27 Vecchione G, Grasselli E, Compalati AD et al. (2016) Ethanol and fatty acids impair lipid homeostasis in an in vitro model of Hepatic Steatosis. Food Chem Toxicol. 90;84-94
28 Wu GY, Rui C, Chen JQ et al. (2017) MicroRNA-122 Inhibits Lipid Droplet Formation and Hepatic Triglyceride Accumulation via Yin Yang 1. Cell Physiol Biochem., 44(4);1651-1664
29 Wong VW, Adams LA, de Lédinghen V et al. (2018) Noninvasive biomarkers in NAFLD and NASH - current progress and future promise. Nat Rev Gastroenterol Hepatol., 15(8);461-478
30 Moylan CA, Pang H, Dellinger A et al., (2014) Hepatic gene expression profiles differentiate presymptomatic patients with mild versus severe nonalcoholic fatty liver disease. Hepatology;59(2):471-82
31 Zhang W, Bu SY, Mashek MT et al., (2016) Integrated Regulation of Hepatic Lipid and Glucose Metabolism by Adipose Triacylglycerol Lipase and FoxO Proteins. Cell Rep. 12:15(2);349-59
Article Sidebar
Article Details
Authors who publish with JBM agree to the following terms:
- Authors retain copyright and grant JBM right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
Saravanakumar Dhakshinamoorthy, Jubilant Biosys Limited
Vice President and Head
Discovery Biology
