A Familial Hypercholesterolemia Human Liver Chimeric Mouse Model Using Induced Pluripotent Stem Cell-derived Hepatocytes

Jiayin Yang; Lai-Yung Wong; Xiao-Yu Tian; Rui Wei; Wing-Hon Lai; Ka-Wing Au; Zhiwei Luo; Carl Ward; Wai-In Ho; David P. Ibañez; Hao Liu; Xichen Bao; Baoming Qin; Yu Huang; Miguel A. Esteban; Hung-Fat Tse

doi:10.3791/57556

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Method Article

A Familial Hypercholesterolemia Human Liver Chimeric Mouse Model Using Induced Pluripotent Stem Cell-derived Hepatocytes

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DOI:

10.3791/57556

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September 15th, 2018

Jiayin Yang^*¹^,²^,⁶ , Lai-Yung Wong^*² , Xiao-Yu Tian^*³ , Rui Wei¹^,² , Wing-Hon Lai² , Ka-Wing Au² , Zhiwei Luo⁴^,⁵ , Carl Ward⁴^,⁵ , Wai-In Ho² , David P. Ibañez⁴^,⁵ , Hao Liu⁴^,⁵ , Xichen Bao⁴^,⁵ , Baoming Qin⁴^,⁵ , Yu Huang³ , Miguel A. Esteban⁴^,⁵^,⁷ , Hung-Fat Tse¹^,²^,⁶^,⁷

¹Department of Medicine, University of Hong Kong-Shenzhen Hospital, ²The Cardiology Division, Department of Medicine, Li Ka Shing Faculty of Medicine, University of Hong Kong, ³School of Biomedical Sciences, Institute of Vascular Medicine, Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, ⁴Key Laboratory of Regenerative Biology of the Chinese Academy of Sciences, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health and Guangzhou Medical University, ⁵Laboratory of RNA, Chromatin, and Human Disease, CAS Key Laboratory of Regenerative Biology and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, ⁶Research Centre of Heart, Brain, Hormone, and Healthy Ageing, Li Ka Shing Faculty of Medicine, University of Hong Kong, ⁷Hong Kong-Guangdong Stem Cell and Regenerative Medicine Research Centre, University of Hong Kong and Guangzhou Institutes of Biomedicine and Health

^* These authors contributed equally

Summary

Here, we present a protocol to generate a human liver chimeric mouse model of familial hypercholesterolemia using human induced pluripotent stem cell-derived hepatocytes. This is a valuable model for testing new therapies for hypercholesterolemia.

Abstract

Familial hypercholesterolemia (FH) is mostly caused by low-density lipoprotein receptor (LDLR) mutations and results in an increased risk of early-onset cardiovascular disease due to marked elevation of LDL cholesterol (LDL-C) in blood. Statins are the first line of lipid-lowering drugs for treating FH and other types of hypercholesterolemia, but new approaches are emerging, in particular PCSK9 antibodies, which are now being tested in clinical trials. To explore novel therapeutic approaches for FH, either new drugs or new formulations, we need appropriate in vivo models. However, differences in the lipid metabolic profiles compared to humans are a key problem of the available animal models of FH. To address this issue, we have generated a human liver chimeric mouse model using FH induced pluripotent stem cell (iPSC)-derived hepatocytes (iHeps). We used Ldlr^-/-/Rag2^-/-/Il2rg^-/- (LRG) mice to avoid immune rejection of transplanted human cells and to assess the effect of LDLR-deficient iHeps in an LDLR null background. Transplanted FH iHeps could repopulate 5-10% of the LRG mouse liver based on human albumin staining. Moreover, the engrafted iHeps responded to lipid-lowering drugs and recapitulated clinical observations of increased efficacy of PCSK9 antibodies compared to statins. Our human liver chimeric model could thus be useful for preclinical testing of new therapies to FH. Using the same protocol, similar human liver chimeric mice for other FH genetic variants, or mutations corresponding to other inherited liver diseases, may also be generated.

Introduction

Low-density lipoprotein receptor (LDLR) captures LDL cholesterol (LDL-C) in blood to modulate cholesterol synthesis in the liver. Mutations in the LDLR gene are the most frequent cause of familial hypercholesterolemia (FH)¹. Statins have traditionally been the first line of medication to treat FH and other types of hypercholesterolemia (inherited or acquired). Statins inhibit 3-hydroxy-3-methylglutaryl-coenzyme A reductase to lower cholesterol synthesis in the liver². Additionally, statins increase LDLR levels on the hepatocyte surface to promote plasma LDL-C clearance. However, a major caveat of treatment with statins i....