T Cell Mediated Immune Responses as a Regulator of Heart Failure

T 细胞介导的免疫反应作为心力衰竭的调节剂

• 批准号: 8750156
• 负责人: Maria Pilar Alcaide Alonso
• 金额: $ 41.25万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8750156
• 关键词: Adhesions; Affect; Anti-Inflammatory Agents; Anti-inflammatory; Biological Assay; Blood; Blood Vessels; CD4 Positive T Lymphocytes; Cardiac; Cardiac Myocytes; Cardiovascular system; Cell Adhesion Molecules; Cell Communication; Cessation of life; Chest; Chronic; Clinical Trials; Data; Disease; Echocardiography; Endothelial Cells; Fibroblasts; Fibrosis; Flow Cytometry; Goals; Heart; Heart failure; Hospitalization; Human; Human Biology; Immune response; Immunohistochemistry; Immunology; In Vitro; Inflammation; Inflammatory; Integrins; Left; Left Ventricular Dysfunction; Left Ventricular Function; Lymphoid; Mediating; Modeling; Molecular; Mus; Organ; Pathway interactions; Patients; Peripheral; Pharmaceutical Preparations; Physiological; Physiology; Pilot Projects; Play; Process; Recruitment Activity; Regulation; Regulatory T-Lymphocyte; Research; Role; Sampling; Selectins; Source; Staging; Staining method; Stains; Stress; Syndrome; System; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Therapeutic; Time; Translating; Vascular Endothelial Cell; Vascular Endothelium; Ventricular; Video Microscopy; Western Blotting; Work; base; cell mediated immune response; chemokine receptor; constriction; cytokine; hemodynamics; immune activation; improved; in vivo; innovation; migration; mortality; new therapeutic target; novel; outcome forecast; pressure; prevent; public health relevance; reconstitution; response; ventricular assist device

DESCRIPTION (provided by applicant): Heart failure (HF) is a progressive syndrome affecting nearly 20 million people worldwide and is generally caused by remodeling of the heart in response to pathological stress. Emerging evidence associates systemic inflammation with left ventricular (LV) dysfunction and HF, but the mechanisms for the observed systemic inflammation are largely unexplored. Our preliminary data using Thoracic aortic constriction (TAC), a well-defined model of HF, indicates that T cells are recruited to the heart and the heart vascular endothelium is activated locally as LV function worsens. Interestingly, we found that T cells from mice undergoing HF interact with activated mouse heart endothelial cells in significant higher numbers than T cells from control mice under physiological flow conditions in vitro. Remarkably, T cell deficient mice (TCR¿-/-) showed increased survival, preserved LV function, and decreased fibrosis in our pilot studies. Based on these data, in this proposal we will test the central hypothesis that T cell mediated immune responses influence cardiac remodeling in pressure overload induced HF. We propose 3 specific aims to test this hypothesis, in which we will obtain novel information about the mechanisms regulating mouse and human T cell recruitment in the heart during the course of HF, the T cell subsets involved, and their influence in the mechanisms regulating cardiac remodeling in HF. In Aim 1 we will use videomicroscopy to study the adhesion mechanisms regulating interactions between mouse T cells and heart endothelial cells during the progression of HF induced by TAC, determining if T cells from mice with HF utilize selectin and/or integrin/Ig superfamily pathways, to mediate rolling, arrest and transendothelial migration under physiological flow conditions in vitro. The expression and functionality of adhesion molecules and chemokine receptors in T cells from mice with HF will also be characterized by flow cytometry. Aim 2 will characterize the T cell mediated immune response induced by TAC, define the role of T cell subsets during the progression of HF, and their role in cardiomyocyte and cardiac fibroblast function in vitro. TAC studies will be performed in WT and TCR¿-/- mice and different T cell subsets will be adoptively transferred into TCR¿-/- recipient mice undergoing TAC. LV function will be determined by echocardiography and hemodynamic studies, fibrosis by picosirious red staining, and flow cytometry, qPCR, western blot and Immunohistochemistry approaches will be used to determine the effect of T cell subsets and their cytokines in cardiac myocyte and fibroblast function. Similar approaches will be used in Aim 3 to translate our findings in Aims 1 and 2 to human biology by exploring the mechanisms regulating human T cell recruitment in HF. Completion of these aims will result in a deeper understanding of how best to regulate T cell mediated inflammation to improve the structural, functional and molecular deficits of the failing heart, and will identify alternative therapeutic approaches to treat HF based on our basic understanding of novel mechanisms that control the timing, type and progression of the T cell immune response in pathological remodeling of the heart.

描述（由申请人提供）：心力衰竭 (HF) 是一种进行性综合征，影响全世界近 2000 万人，通常是由心脏对病理应激做出的重塑引起的。新出现的证据将全身炎症与左心室 (LV) 功能障碍和心力衰竭联系起来，但观察到的全身炎症的机制很大程度上尚未被探索。我们使用胸主动脉缩窄（TAC）（一种明确的心力衰竭模型）的初步数据表明，随着左心室功能恶化，T细胞被募集到心脏，并且心脏血管内皮被局部激活。有趣的是，我们发现在体外生理流动条件下，经历心力衰竭的小鼠的 T 细胞与激活的小鼠心脏内皮细胞相互作用的数量显着高于对照小鼠的 T 细胞。值得注意的是，在我们的初步研究中，T 细胞缺陷小鼠 (TCR¿-/-) 表现出存活率增加、左心室功能得以保留以及纤维化减少。基于这些数据，在本提案中我们将测试 中心假设是 T 细胞介导的免疫反应影响压力超负荷引起的心力衰竭的心脏重塑。我们提出了 3 个具体目标来检验这一假设，其中我们将获得有关心力衰竭过程中调节小鼠和人类心脏 T 细胞募集的机制、涉及的 T 细胞亚群及其对调节心力衰竭心脏重塑机制的影响的新信息。在目标 1 中，我们将使用视频显微镜研究在 TAC 诱导的心力衰竭进展过程中调节小鼠 T 细胞和心脏内皮细胞之间相互作用的粘附机制，确定心力衰竭小鼠的 T 细胞是否利用选择素和/或整合素/Ig 超家族途径，在体外生理流动条件下介导滚动、阻滞和跨内皮迁移。心衰小鼠 T 细胞中粘附分子和趋化因子受体的表达和功能也将通过流式细胞术进行表征。目标 2 将表征 TAC 诱导的 T 细胞介导的免疫反应，定义 T 细胞亚群在心力衰竭进展过程中的作用，及其在体外心肌细胞和心脏成纤维细胞功能中的作用。将进行 TAC 研究 在WT和TCR¿-/-小鼠中，不同的T细胞亚群将被过继转移到接受TAC的TCR¿-/-受体小鼠中。左心室功能将通过超声心动图和血流动力学研究确定，纤维化通过皮可西里红染色确定，流式细胞术、qPCR、蛋白质印迹和免疫组织化学方法将用于确定 T 细胞亚群及其细胞因子对心肌细胞和成纤维细胞功能的影响。类似的方法将用于目标 3，通过探索心力衰竭中调节人类 T 细胞募集的机制，将我们在目标 1 和 2 中的发现转化为人类生物学。完成这些目标将导致我们更深入地了解如何最好地调节 T 细胞介导的炎症，以改善衰竭心脏的结构、功能和分子缺陷，并将基于我们对控制心脏病理重塑中 T 细胞免疫反应的时间、类型和进展的新机制的基本理解，确定治疗心力衰竭的替代治疗方法。