Engineered Regulatory T Lymphocytes Promote Infarcted Heart Repair
Abstract
BACKGROUND: Myocardial infarction (MI) initiates a dysregulated healing process characterized by excessive fibrosis and unresolved inflammation, resulting in suboptimal cardiac repair in clinical settings. Regulatory T lymphocytes (Tregs) naturally orchestrate cardiac repair after MI, but their therapeutic potential is limited by inefficient homing to ischemic myocardium. We hypothesize that FAP (fibroblast activation protein)–specific CAR (chimeric antigen receptor) engineering overcomes this barrier by enabling precise delivery of Tregs to FAP⁺-enriched infarct zones, thereby focally amplifying reparative activity within injured myocardium. METHODS: In murine MI and ischemia–reperfusion models, C57BL/6J mice were injected with lentivirus-engineered FAP CAR Tregs (FCTRs) or mock Tregs derived from wild-type, IL-10 (interleukin-10) knockout ( IL-10 −/− ) or Areg (amphiregulin) knockout ( Areg −/− ) donors after infarction. The cardiac outcomes and underlying mechanisms mediated by FCTRs were thoroughly analyzed. Systemic toxicity was evaluated to ensure safety. RESULTS: Intravenous injections of FCTRs on day 3 after injury led to targeted engraftment in the damaged cardiac tissue. Compared with controls treated with vehicle or mock Tregs, mice receiving FCTRs exhibited remarkable cardiac functional recovery in both MI and ischemia–reperfusion models by day 14, accompanied by reduced fibrosis and decreased inflammation, all achieved without compromising the integrity of cardiac tissue. Absence of IL-10 in the engineered CAR Tregs abrogated their therapeutic efficacy, whereas the ablation of Areg showed no functional impairment. We further demonstrated that the beneficial effects of FCTRs depended on IL-10 production, which inhibited pathogenic myofibroblast differentiation by suppressing Smad2/3-dependent signaling. In addition, IL-10 secretion by these engineered Tregs promoted the polarization of inflammatory monocytes into reparative M2 macrophages and resolved excessive inflammatory responses. No treatment-related adverse effects were observed. CONCLUSIONS: We pioneered FAP-targeted CAR Tregs as a dual-action precision therapy resolving post-MI fibrosis and inflammation through IL-10–dependent mechanisms. By spatiotemporally suppressing myofibroblast differentiation and remodeling immune niches, this strategy prevents maladaptive remodeling while accelerating functional recovery, establishing a translational platform for fibrotic diseases across organ systems.
Article Details
Authors (19)
Min Zhang
Yongying Qin
Department of Cardiology, Hubei Key Laboratory of Biological Targeted Therapy, Hubei Provincial Engineering Research Center of Immunological Diagnosis and Therapy for Cardiovascular Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (M.Z., Y.Y.Q., T.Z., M.L.L., T.T.T., N.X., S.F.N., B.J.L., Z.F.Z., J.J., M.Y.G., J.Y.L., X.C.).
Ting Zhou
Meilin Liu
School of Materials Science and Engineering
Tingting Tang
Ni Xia
Department of Cardiology, Hubei Key Laboratory of Biological Targeted Therapy, Hubei Provincial Engineering Research Center of Immunological Diagnosis and Therapy for Cardiovascular Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (M.Z., Y.Y.Q., T.Z., M.L.L., T.T.T., N.X., S.F.N., B.J.L., Z.F.Z., J.J., M.Y.G., J.Y.L., X.C.).
Shaofang Nie
Department of Cardiology, Hubei Key Laboratory of Biological Targeted Therapy, Hubei Provincial Engineering Research Center of Immunological Diagnosis and Therapy for Cardiovascular Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (M.Z., Y.Y.Q., T.Z., M.L.L., T.T.T., N.X., S.F.N., B.J.L., Z.F.Z., J.J., M.Y.G., J.Y.L., X.C.).
Bingjie Lv
Department of Cardiology, Hubei Key Laboratory of Biological Targeted Therapy, Hubei Provincial Engineering Research Center of Immunological Diagnosis and Therapy for Cardiovascular Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (M.Z., Y.Y.Q., T.Z., M.L.L., T.T.T., N.X., S.F.N., B.J.L., Z.F.Z., J.J., M.Y.G., J.Y.L., X.C.).
Zhengfeng Zhu
Jiao Jiao
School of Chemistry, Institute of New Concept Sensors and Molecular Materials (INCSMM), State Key Laboratory of Fluorine & Nitrogen Chemicals, Engineering Research Center of Energy Storage Materials and Devices, Ministry of Education, Xi’an Key Laboratory of Sustainable Polymer Materials
Muyang Gu
Department of Cardiology, Hubei Key Laboratory of Biological Targeted Therapy, Hubei Provincial Engineering Research Center of Immunological Diagnosis and Therapy for Cardiovascular Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China (M.Z., Y.Y.Q., T.Z., M.L.L., T.T.T., N.X., S.F.N., B.J.L., Z.F.Z., J.J., M.Y.G., J.Y.L., X.C.).
Jingyong Li
Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology
Chen Chen
Desheng Hu
Weimin Wang
MAX IV Laboratory, Fotongatan 2, Lund, SE-22484, Sweden
Li Zhang
Chaolong Wang
Zhilei Shan
Department of Nutrition and Food Hygiene (Z.L.S.), School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Xiang Cheng