Heterogeneity of Hepatic Stellate Cells in Fibrogenesis of the Liver: Insights from Single-Cell Transcriptomic Analysis in Liver Injury
Background & Aims: Liver fibrosis is a pathological healing process resulting from hepatic stellate cell (HSC) activation and the generation of myofibroblasts from activated HSCs. The precise underlying mechanisms of liver fibrogenesis are still largely vague due to lack of understanding the fun...
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MDPI AG
2021-08-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/10/8/2129 |
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| author | Wenjun Zhang Simon J. Conway Ying Liu Paige Snider Hanying Chen Hongyu Gao Yunlong Liu Kadir Isidan Kevin J. Lopez Gonzalo Campana Ping Li Burcin Ekser Heather Francis Weinian Shou Chandrashekhar Kubal |
| author_facet | Wenjun Zhang Simon J. Conway Ying Liu Paige Snider Hanying Chen Hongyu Gao Yunlong Liu Kadir Isidan Kevin J. Lopez Gonzalo Campana Ping Li Burcin Ekser Heather Francis Weinian Shou Chandrashekhar Kubal |
| author_sort | Wenjun Zhang |
| collection | DOAJ |
| description | Background & Aims: Liver fibrosis is a pathological healing process resulting from hepatic stellate cell (HSC) activation and the generation of myofibroblasts from activated HSCs. The precise underlying mechanisms of liver fibrogenesis are still largely vague due to lack of understanding the functional heterogeneity of activated HSCs during liver injury. Approach and Results: In this study, to define the mechanism of HSC activation, we performed the transcriptomic analysis at single-cell resolution (scRNA-seq) on HSCs in mice treated with carbon tetrachloride (CCl<sub>4</sub>). By employing <i>LRAT-Cre:Rosa26<sup>mT/mG</sup></i> mice, we were able to isolate an activated GFP-positive HSC lineage derived cell population by fluorescence-activated cell sorter (FACS). A total of 8 HSC subpopulations were identified based on an unsupervised analysis. Each HSC cluster displayed a unique transcriptomic profile, despite all clusters expressing common mouse HSC marker genes. We demonstrated that one of the HSC subpopulations expressed high levels of mitosis regulatory genes, velocity, and monocle analysis indicated that these HSCs are at transitioning and proliferating phases at the beginning of HSCs activation and will eventually give rise to several other HSC subtypes. We also demonstrated cell clusters representing HSC-derived mature myofibroblast populations that express myofibroblasts hallmark genes with unique contractile properties. Most importantly, we found a novel HSC cluster that is likely to be critical in liver regeneration, immune reaction, and vascular remodeling, in which the unique profiles of genes such as Rgs5, Angptl6, and Meg3 are highly expressed. Lastly, we demonstrated that the heterogeneity of HSCs in the injured mouse livers is closely similar to that of cirrhotic human livers. Conclusions: Collectively, our scRNA-seq data provided insight into the landscape of activated HSC populations and the dynamic transitional pathway from HSC to myofibroblasts in response to liver injury. |
| first_indexed | 2024-03-10T08:55:18Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T08:55:18Z |
| publishDate | 2021-08-01 |
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| spelling | doaj.art-2917a3cd6c98492e959c27df7bac150d2023-11-22T07:12:10ZengMDPI AGCells2073-44092021-08-01108212910.3390/cells10082129Heterogeneity of Hepatic Stellate Cells in Fibrogenesis of the Liver: Insights from Single-Cell Transcriptomic Analysis in Liver InjuryWenjun Zhang0Simon J. Conway1Ying Liu2Paige Snider3Hanying Chen4Hongyu Gao5Yunlong Liu6Kadir Isidan7Kevin J. Lopez8Gonzalo Campana9Ping Li10Burcin Ekser11Heather Francis12Weinian Shou13Chandrashekhar Kubal14Division of Transplant Surgery, Indiana University, Indianapolis, IN 46202, USADepartment of Pediatrics, Indiana University School of Medicine, Indianapolis, IN 46202, USADepartment of Pediatrics, Indiana University School of Medicine, Indianapolis, IN 46202, USADepartment of Pediatrics, Indiana University School of Medicine, Indianapolis, IN 46202, USAGenome Editing Center, Indiana University School of Medicine, Indianapolis, IN 46202, USAThe Center for Medical Genomics, Indiana University School of Medicine, Indianapolis, IN 46202, USAThe Center for Medical Genomics, Indiana University School of Medicine, Indianapolis, IN 46202, USADivision of Transplant Surgery, Indiana University, Indianapolis, IN 46202, USADivision of Transplant Surgery, Indiana University, Indianapolis, IN 46202, USADivision of Transplant Surgery, Indiana University, Indianapolis, IN 46202, USADivision of Transplant Surgery, Indiana University, Indianapolis, IN 46202, USADivision of Transplant Surgery, Indiana University, Indianapolis, IN 46202, USADivision of Gastroenterology, Department of Medicine, Indiana University School of Medicine, Indianapolis, IN 46202, USADepartment of Pediatrics, Indiana University School of Medicine, Indianapolis, IN 46202, USADivision of Transplant Surgery, Indiana University, Indianapolis, IN 46202, USABackground & Aims: Liver fibrosis is a pathological healing process resulting from hepatic stellate cell (HSC) activation and the generation of myofibroblasts from activated HSCs. The precise underlying mechanisms of liver fibrogenesis are still largely vague due to lack of understanding the functional heterogeneity of activated HSCs during liver injury. Approach and Results: In this study, to define the mechanism of HSC activation, we performed the transcriptomic analysis at single-cell resolution (scRNA-seq) on HSCs in mice treated with carbon tetrachloride (CCl<sub>4</sub>). By employing <i>LRAT-Cre:Rosa26<sup>mT/mG</sup></i> mice, we were able to isolate an activated GFP-positive HSC lineage derived cell population by fluorescence-activated cell sorter (FACS). A total of 8 HSC subpopulations were identified based on an unsupervised analysis. Each HSC cluster displayed a unique transcriptomic profile, despite all clusters expressing common mouse HSC marker genes. We demonstrated that one of the HSC subpopulations expressed high levels of mitosis regulatory genes, velocity, and monocle analysis indicated that these HSCs are at transitioning and proliferating phases at the beginning of HSCs activation and will eventually give rise to several other HSC subtypes. We also demonstrated cell clusters representing HSC-derived mature myofibroblast populations that express myofibroblasts hallmark genes with unique contractile properties. Most importantly, we found a novel HSC cluster that is likely to be critical in liver regeneration, immune reaction, and vascular remodeling, in which the unique profiles of genes such as Rgs5, Angptl6, and Meg3 are highly expressed. Lastly, we demonstrated that the heterogeneity of HSCs in the injured mouse livers is closely similar to that of cirrhotic human livers. Conclusions: Collectively, our scRNA-seq data provided insight into the landscape of activated HSC populations and the dynamic transitional pathway from HSC to myofibroblasts in response to liver injury.https://www.mdpi.com/2073-4409/10/8/2129single-cell RNA sequencinghepatic stellate cell sublineagemyofibroblastliver fibrosiscarbon tetrachloride |
| spellingShingle | Wenjun Zhang Simon J. Conway Ying Liu Paige Snider Hanying Chen Hongyu Gao Yunlong Liu Kadir Isidan Kevin J. Lopez Gonzalo Campana Ping Li Burcin Ekser Heather Francis Weinian Shou Chandrashekhar Kubal Heterogeneity of Hepatic Stellate Cells in Fibrogenesis of the Liver: Insights from Single-Cell Transcriptomic Analysis in Liver Injury Cells single-cell RNA sequencing hepatic stellate cell sublineage myofibroblast liver fibrosis carbon tetrachloride |
| title | Heterogeneity of Hepatic Stellate Cells in Fibrogenesis of the Liver: Insights from Single-Cell Transcriptomic Analysis in Liver Injury |
| title_full | Heterogeneity of Hepatic Stellate Cells in Fibrogenesis of the Liver: Insights from Single-Cell Transcriptomic Analysis in Liver Injury |
| title_fullStr | Heterogeneity of Hepatic Stellate Cells in Fibrogenesis of the Liver: Insights from Single-Cell Transcriptomic Analysis in Liver Injury |
| title_full_unstemmed | Heterogeneity of Hepatic Stellate Cells in Fibrogenesis of the Liver: Insights from Single-Cell Transcriptomic Analysis in Liver Injury |
| title_short | Heterogeneity of Hepatic Stellate Cells in Fibrogenesis of the Liver: Insights from Single-Cell Transcriptomic Analysis in Liver Injury |
| title_sort | heterogeneity of hepatic stellate cells in fibrogenesis of the liver insights from single cell transcriptomic analysis in liver injury |
| topic | single-cell RNA sequencing hepatic stellate cell sublineage myofibroblast liver fibrosis carbon tetrachloride |
| url | https://www.mdpi.com/2073-4409/10/8/2129 |
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