학술논문
Liver specification of human iPSC-derived endothelial cells transplanted into mouse liver.
Document Type
Academic Journal
Author
Yap KK; O'Brien Department of St Vincent's Institute, Fitzroy, VIC, Australia.; University of Melbourne Department of Surgery, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.; Schröder J; Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC, Australia.; Monash Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia.; Australian Regenerative Medicine Institute, Clayton, VIC, Australia.; Doherty Institute & University of Melbourne Department of Microbiology and Immunology, Parkville, VIC, Australia.; Gerrand YW; O'Brien Department of St Vincent's Institute, Fitzroy, VIC, Australia.; Dobric A; O'Brien Department of St Vincent's Institute, Fitzroy, VIC, Australia.; Kong AM; O'Brien Department of St Vincent's Institute, Fitzroy, VIC, Australia.; Fox AM; University of Melbourne Department of Surgery, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.; Hepatobiliary Surgery Unit, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.; Knowles B; University of Melbourne Department of Surgery, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.; Hepatobiliary Surgery Unit, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.; Banting SW; University of Melbourne Department of Surgery, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.; Hepatobiliary Surgery Unit, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.; Elefanty AG; Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC, Australia.; Murdoch Children's Research Institute, The Royal Children's Hospital, Flemington Road, Parkville, VIC, Australia.; Department of Paediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, Australia.; Stanley EG; Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC, Australia.; Murdoch Children's Research Institute, The Royal Children's Hospital, Flemington Road, Parkville, VIC, Australia.; Department of Paediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, VIC, Australia.; Yeoh GC; Harry Perkins Institute of Medical Research and Centre for Medical Research, University of Western Australia, Perth, WA, Australia.; Lockwood GP; ANZAC Research Institute and University of Sydney, Concord, NSW, Australia.; Cogger VC; ANZAC Research Institute and University of Sydney, Concord, NSW, Australia.; Morrison WA; O'Brien Department of St Vincent's Institute, Fitzroy, VIC, Australia.; University of Melbourne Department of Surgery, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.; Australian Catholic University, Fitzroy, VIC, Australia.; Polo JM; Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC, Australia.; Monash Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia.; Australian Regenerative Medicine Institute, Clayton, VIC, Australia.; Adelaide Centre for Epigenetics, South Australian Immunogenomics Cancer Institute, University of Adelaide, Adelaide, SA, Australia.; Mitchell GM; O'Brien Department of St Vincent's Institute, Fitzroy, VIC, Australia.; University of Melbourne Department of Surgery, St Vincent's Hospital Melbourne, Fitzroy, VIC, Australia.; Australian Catholic University, Fitzroy, VIC, Australia.
Source
Publisher: Elsevier B.V Country of Publication: Netherlands NLM ID: 101761237 Publication Model: eCollection Cited Medium: Internet ISSN: 2589-5559 (Electronic) Linking ISSN: 25895559 NLM ISO Abbreviation: JHEP Rep Subsets: PubMed not MEDLINE
Subject
Language
English
Abstract
Background & Aims: Liver sinusoidal endothelial cells (LSECs) are important in liver development, regeneration, and pathophysiology, but the differentiation process underlying their tissue-specific phenotype is poorly understood and difficult to study because primary human cells are scarce. The aim of this study was to use human induced pluripotent stem cell (hiPSC)-derived LSEC-like cells to investigate the differentiation process of LSECs.
Methods: hiPSC-derived endothelial cells (iECs) were transplanted into the livers of Fah -/- / Rag2 -/- / Il2rg -/- mice and assessed over a 12-week period. Lineage tracing, immunofluorescence, flow cytometry, plasma human factor VIII measurement, and bulk and single cell transcriptomic analysis were used to assess the molecular and functional changes that occurred following transplantation.
Results: Progressive and long-term repopulation of the liver vasculature occurred as iECs expanded along the sinusoids between hepatocytes and increasingly produced human factor VIII, indicating differentiation into LSEC-like cells. To chart the developmental profile associated with LSEC specification, the bulk transcriptomes of transplanted cells between 1 and 12 weeks after transplantation were compared against primary human adult LSECs. This demonstrated a chronological increase in LSEC markers, LSEC differentiation pathways, and zonation. Bulk transcriptome analysis suggested that the transcription factors NOTCH1 , GATA4 , and FOS have a central role in LSEC specification, interacting with a network of 27 transcription factors. Novel markers associated with this process included EMCN and CLEC14A . Additionally, single cell transcriptomic analysis demonstrated that transplanted iECs at 4 weeks contained zonal subpopulations with a region-specific phenotype.
Conclusions: Collectively, this study confirms that hiPSCs can adopt LSEC-like features and provides insight into LSEC specification. This humanised xenograft system can be applied to further interrogate LSEC developmental biology and pathophysiology, bypassing current logistical obstacles associated with primary human LSECs.
Impact and Implications: Liver sinusoidal endothelial cells (LSECs) are important cells for liver biology, but better model systems are required to study them. We present a pluripotent stem cell xenografting model that produces human LSEC-like cells. A detailed and longitudinal transcriptomic analysis of the development of LSEC-like cells is included, which will guide future studies to interrogate LSEC biology and produce LSEC-like cells that could be used for regenerative medicine.
Competing Interests: None to declare. Please refer to the accompanying ICMJE disclosure forms for further details.
(© 2024 The Author(s).)
Methods: hiPSC-derived endothelial cells (iECs) were transplanted into the livers of Fah
Results: Progressive and long-term repopulation of the liver vasculature occurred as iECs expanded along the sinusoids between hepatocytes and increasingly produced human factor VIII, indicating differentiation into LSEC-like cells. To chart the developmental profile associated with LSEC specification, the bulk transcriptomes of transplanted cells between 1 and 12 weeks after transplantation were compared against primary human adult LSECs. This demonstrated a chronological increase in LSEC markers, LSEC differentiation pathways, and zonation. Bulk transcriptome analysis suggested that the transcription factors NOTCH1 , GATA4 , and FOS have a central role in LSEC specification, interacting with a network of 27 transcription factors. Novel markers associated with this process included EMCN and CLEC14A . Additionally, single cell transcriptomic analysis demonstrated that transplanted iECs at 4 weeks contained zonal subpopulations with a region-specific phenotype.
Conclusions: Collectively, this study confirms that hiPSCs can adopt LSEC-like features and provides insight into LSEC specification. This humanised xenograft system can be applied to further interrogate LSEC developmental biology and pathophysiology, bypassing current logistical obstacles associated with primary human LSECs.
Impact and Implications: Liver sinusoidal endothelial cells (LSECs) are important cells for liver biology, but better model systems are required to study them. We present a pluripotent stem cell xenografting model that produces human LSEC-like cells. A detailed and longitudinal transcriptomic analysis of the development of LSEC-like cells is included, which will guide future studies to interrogate LSEC biology and produce LSEC-like cells that could be used for regenerative medicine.
Competing Interests: None to declare. Please refer to the accompanying ICMJE disclosure forms for further details.
(© 2024 The Author(s).)