Xiumeng Hua1, Gang Hu2, Qingtao Hu3, Yuan Chang4, Yiqing Hu4, Linlin Gao3, Xiao Chen1, Ping-Chang Yang5, Yu Zhang6, Mingyao Li7, Jiangping Song1. 1. State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China. 2. School of Statistics and Data Science, Key Laboratory for medical Data Analysis and Statistical Research of Tianjin, Nankai University, Tianjin, China. 3. National Institute of Biological Sciences, Beijing, China. 4. State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China; Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China. 5. Research Center of Allergy and Immunology, Shenzhen University School of Medicine, Shenzhen, Guangzhou, China. 6. National Institute of Biological Sciences, Beijing, China; Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua University, Beijing, China. 7. Department of Biostatistics and Epidemiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA.
Abstract
Background: Myocarditis can develop into dilated cardiomyopathy (DCM), which may require heart transplantation (HTx). The immunological network of myocarditis phases remains unknown. This study aimed to investigate the immunological network during the transition from myocarditis to cardiomyopathy and to identify the genes contributing to the inflammatory response to myocarditis. Methods: Mice were treated with myosin heavy-chain-α peptides to generate an experimental autoimmune myocarditis (EAM) model. We performed single-cell RNA sequencing (scRNA-seq) analysis of Cd45+ cells extracted from mouse hearts during different EAM phases, including normal control, acute inflammatory, subacute inflammatory and myopathy phases. Human heart tissues were collected from the surgically removed hearts of patients who had undergone HTx. Results: We identified 26 cell subtypes among 34,665 cells. Macrophages constituted the main immune cell population at all disease phases (greater than 60%), and an inflammation-associated macrophage cluster was identified in which the expression of Hif1a-regulated genes was upregulated. The neutrophil population was increased after the induction of EAM, and neutrophils then released Il-1 to participate in the EAM process. T cells were observed at the highest percentage at the subacute inflammatory phase. Th17 cells, in which the expression of Hif1a-regulated genes was upregulated, constituted the main T cell population detected at the acute inflammatory phase, while Treg cells were the main T cell population detected at the subacute inflammatory phase, and γδ T cells releasing Il-17 were the main T cell population observed at the myopathy phase. Moreover, the Hif1a expression level correlated with the extent of inflammation. Additionally, PX-478 could alleviate the inflammatory responses of the different EAM phases. Finally, HIF1A was expressed at higher levels in acute autoimmune myocarditis patients than in DCM patients and healthy controls. Conclusions: We herein present a comprehensive single-cell landscape of the cardiac immune cells in different EAM phases. In addition, we elucidated the contribution of Hif1a to the inflammatory response through the regulation of immune cell activity, particularly of macrophage cluster 2 and Th17 cells. Moreover, a Hif1a inhibitor alleviated inflammatory cell infiltration of the EAM model and may serve as a potential therapeutic target in the clinic.
Background: Myocarditis can develop into dilated cardiomyopathy (DCM), which may require heart transplantation (HTx). The immunological network of myocarditis phases remains unknown. This study aimed to investigate the immunological network during the transition from myocarditis to cardiomyopathy and to identify the genes contributing to the inflammatory response to myocarditis. Methods:Mice were treated with myosin heavy-chain-α peptides to generate an experimental autoimmune myocarditis (EAM) model. We performed single-cell RNA sequencing (scRNA-seq) analysis of Cd45+ cells extracted from mouse hearts during different EAM phases, including normal control, acute inflammatory, subacute inflammatory and myopathy phases. Human heart tissues were collected from the surgically removed hearts of patients who had undergone HTx. Results: We identified 26 cell subtypes among 34,665 cells. Macrophages constituted the main immune cell population at all disease phases (greater than 60%), and an inflammation-associated macrophage cluster was identified in which the expression of Hif1a-regulated genes was upregulated. The neutrophil population was increased after the induction of EAM, and neutrophils then released Il-1 to participate in the EAM process. T cells were observed at the highest percentage at the subacute inflammatory phase. Th17 cells, in which the expression of Hif1a-regulated genes was upregulated, constituted the main T cell population detected at the acute inflammatory phase, while Treg cells were the main T cell population detected at the subacute inflammatory phase, and γδ T cells releasing Il-17 were the main T cell population observed at the myopathy phase. Moreover, the Hif1a expression level correlated with the extent of inflammation. Additionally, PX-478 could alleviate the inflammatory responses of the different EAM phases. Finally, HIF1A was expressed at higher levels in acute autoimmune myocarditispatients than in DCM patients and healthy controls. Conclusions: We herein present a comprehensive single-cell landscape of the cardiac immune cells in different EAM phases. In addition, we elucidated the contribution of Hif1a to the inflammatory response through the regulation of immune cell activity, particularly of macrophage cluster 2 and Th17 cells. Moreover, a Hif1a inhibitor alleviated inflammatory cell infiltration of the EAM model and may serve as a potential therapeutic target in the clinic.
Authors: Daniel J Tyrrell; Judy Chen; Benjamin Y Li; Sherri C Wood; Wendy Rosebury-Smith; Henriette A Remmer; Longtan Jiang; Min Zhang; Morgan Salmon; Gorav Ailawadi; Bo Yang; Daniel R Goldstein Journal: Arterioscler Thromb Vasc Biol Date: 2022-05-05 Impact factor: 10.514