Project 2 - Role of Inflammation in CMC-Induced Myocardal Repair

项目 2 - 炎症在 CMC 诱导的心肌修复中的作用

• 批准号: 9359611
• 负责人: Marcin Wysoczynski
• 金额: $ 33.12万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-15 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9359611
• 关键词: Affect; Alpha Cell; Bone Marrow; Cardiac; Cardiac Myocytes; Cardiovascular system; Cell Therapy; Cells; Cessation of life; Clinical; Clinical Research; Coculture Techniques; Data; Diabetes Mellitus; Disease; Fibrosis; Gene Expression; Genetic; Heart; Heart failure; Homeostasis; Homo; Immune; Immune response; Immune system; In Vitro; Infiltration; Inflammation; Inflammatory; Injectable; Injection of therapeutic agent; Injury; Knowledge; Left; Left Ventricular Function; Left ventricular structure; Mediating; Mediator of activation protein; Mesenchymal; MicroRNAs; Mus; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardium; Natural regeneration; Nature; Neonatal; Outcome; Patients; Phenotype; Play; Population; Process; Proliferating; Pump; Recruitment Activity; Role; Rupture; Signal Transduction; Spleen; Sterility; Surface; TNFRSF5 gene; Testing; Time; Tissues; Transgenic Organisms; Ventricular; angiogenesis; base; cardiac repair; cell type; extracellular vesicles; improved; in vivo; inhibitor/antagonist; insight; macrophage; monocyte; myocardial damage; novel; p65; preclinical study; programs; repaired; response; synergism

The mechanism by which cell therapy improves cardiac function remains unclear. Although injected cells minimally differentiate into cardiomyocytes or vessels, and only a small fraction survive long term in the recipient, common beneficial effects are observed regardless of injected cell type: improvement in pump function, reduction of fibrosis, and enhanced angiogenesis. These results are consistent with the idea that cell therapy recruits endogenous repair mechanisms which, however, have not been identified. The immune system has been implicated in a variety of sterile diseases, including processes regulating myocardial damage and repair. Although unresolved inflammatory processes controlled by infiltrating and tissue-resident immune cells worsen heart failure, depletion of macrophages in the infarcted myocardium leads to LV rupture and death. Thus, immune cells appear to play diametrically opposite roles in the heart. Macrophages have been shown to be required for spontaneous regeneration of neonatal mammalian myocardium after injury, and recent findings implicate them as direct contributors to cell therapy-mediated myocardial repair. Nevertheless, how immune cells regulate myocardial repair and what determines their harmful versus salutary actions remains unknown. Furthermore, the impact of cell therapy on reparative immune cells has not yet been studied in the heart. Our preliminary data show that injection of cardiac mesenchymal cells (CMCs) into the infarcted heart promotes accumulation of reparative macrophages. Thus, the central hypothesis of this proposal is that CMCs facilitate recruitment of monocytes and activation of reparative macrophages, which are essential endogenous mediators of repair. By generating detailed flow cytometric analyses of immune cell populations following CMC administration, we will not only resolve the time course of immune cell recruitment, but also determine how inflammation is eventually extinguished in the heart after cell therapy. To elucidate the mechanism whereby CMCs regulate inflammatory processes in monocyte-derived macrophages, we will determine how these cells regulate NFB-p65 subunit expression, with emphasis on horizontal transfer of miRNAs to macrophages through CMC-derived EVs. Finally, using macrophage genetic fate mapping and genetically modified CMCs, we will elucidate the role of monocyte-derived macrophages in CMC-induced myocardial repair. This project will be the first systematic analysis of how cell therapy modulates immune cells – a mechanism that has been relatively understudied. The results will provide novel insights not only into the mechanisms regulating cell therapy-mediated myocardial repair, but also into how endogenous reparative activities of macrophages are recruited. We will also determine whether EVs recapitulate the salutary effects of CMCs on immune cells. Thus, these studies have far-reaching implications for our understanding of how the immune system regulates myocardial homeostasis in general.

细胞疗法改善心脏功能的机制尚不清楚。虽然注射的细胞 最低限度地分化成心肌细胞或血管，只有一小部分在心肌细胞中长期存活。 接受者，无论注射的细胞类型如何，都观察到共同的有益效果： 功能，减少纤维化和增强血管生成。这些结果与细胞 然而，治疗招募了尚未确定的内源性修复机制。免疫 该系统与多种不育性疾病有关，包括调节心肌损伤的过程 和修复。尽管未解决的炎症过程由浸润和组织驻留免疫控制， 细胞恶化心力衰竭，梗死心肌中巨噬细胞的耗竭导致LV破裂， 死亡因此，免疫细胞似乎在心脏中发挥完全相反的作用。宏观经济已经 显示为新生哺乳动物心肌损伤后自发再生所需，和 最近的发现暗示它们是细胞疗法介导的心肌修复的直接贡献者。然而，尽管如此， 免疫细胞如何调节心肌修复，以及是什么决定了它们的有害作用与有益作用 仍然未知。此外，细胞疗法对修复性免疫细胞的影响尚未研究 在心脏。我们的初步数据表明，将心肌间充质细胞（CMC）注射到梗死灶中， 心脏促进修复性巨噬细胞的积累。因此，这一提议的核心假设是， CMC促进单核细胞的募集和修复性巨噬细胞的活化，这是必不可少的。 内源性修复介质。通过对免疫细胞群体进行详细的流式细胞术分析， CMC给药后，我们不仅将解决免疫细胞募集的时间进程， 确定细胞治疗后炎症最终如何在心脏中消失。阐明本 CMC调节单核细胞衍生的巨噬细胞中炎症过程的机制，我们将 确定这些细胞如何调节NF-κ B-p65亚基表达，重点是水平转移， miRNA通过CMC衍生的EV进入巨噬细胞。最后，使用巨噬细胞遗传命运作图和 我们将阐明单核细胞衍生的巨噬细胞在CMC诱导的 心肌修复。该项目将是第一个系统分析细胞疗法如何调节免疫细胞 - 这一机制相对来说研究得还不够。结果将提供新的见解，不仅对 调节细胞治疗介导的心肌修复的机制，而且还进入如何内源性修复 巨噬细胞的活性被募集。我们还将确定电动汽车是否再现了 免疫细胞上的CMC。因此，这些研究对我们理解人类如何 免疫系统通常调节心肌稳态。