The Role of Exosomes in Mesenchymal Stem Cell-Mediated Enhancement of Cardiac Contractility

外泌体在间充质干细胞介导的心脏收缩力增强中的作用

• 批准号: 9396770
• 负责人: Joshua Mayourian
• 金额: $ 4.17万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-18 至 2021-08-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9396770
• 关键词: Activities of Daily Living; Address; Apoptotic; Biochemistry; Biological Assay; Ca(2+)-Transporting ATPase; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cells; Cellular biology; Clinical; Clinical Trials; Coupling; Data; Dilated Cardiomyopathy; Dimensions; Disease; Electrophysiology (science); Endoplasmic Reticulum; Engineering; Fibrosis; Foundations; Future; Gene Expression; Genetic; Heart; Heart failure; Human; Human Engineering; In Vitro; Ion Channel; L-Type Calcium Channels; Lead; Mediating; Mediator of activation protein; Mesenchymal Stem Cells; MicroRNAs; Molecular; Motivation; Myocardium; National Heart, Lung, and Blood Institute; Output; Paracrine Communication; Patients; Performance; Physicians; Play; Preparation; Pump; Recovery; Research; Risk; Role; Scientist; Signal Transduction; Solid; Systems Biology; Testing; Tissue Engineering; Tissue Model; Tissues; Training; Transplantation; United States National Institutes of Health; angiogenesis; career; coronary fibrosis; design; exosome; human stem cells; immunoregulation; improved; in vitro Model; inhibitor/antagonist; innovation; insight; interest; mRNA Expression; mathematical model; multidisciplinary; mutant; nanoparticle; non-genetic; novel; paracrine; phospholamban; repaired; stem cell biology

PROJECT SUMMARY An emerging therapy for non-ischemic cardiomyopathy involves the delivery of human mesenchymal stem cells (hMSCs). Clinical trials document modest benefits on cardiac contractility, underscoring a need to better understand and exploit the underlying mechanisms governing hMSCs-cardiomyocyte (hCMs) interactome. Recent studies on hMSC-mediated heart therapies demonstrated that paracrine signaling—via secreted factors—is a crucial mediator of reduced cardiac fibrosis and enhanced angiogenesis. Moreover, hMSC paracrine factors have been shown to impact contractility by altering cardiomyocyte ion channel/pump activity. However, these findings fail to identify the key components of the hMSC secretome for enhancing contractility. We propose utilizing three-dimensional human engineered cardiac tissues (hECTs) as an in vitro model to investigate the role of hMSC exosomes in enhancement of cardiac contractility. Our lab recently discovered that hMSCs enhance hECT contractile force predominantly through paracrine signaling, counteracting adverse risks of hMSC-hCM heterocellular coupling. Importantly, we discovered that the exosomal component of the hMSC secretome is necessary and sufficient for hMSC-paracrine mediated enhancement of hECT contractility. Furthermore, by utilizing a systems biology approach and integrating hECT contractile function results with exosomal miRNA data, we predicted exosomal miRNA-21 as a lead candidate responsible for the favorable contractile effects of hMSC paracrine signaling. We later validated with qPCR that miRNA-21 levels are increased in hECTs supplemented with hMSC exosomes and hMSC total conditioned media relative to control, motivating our central hypothesis that exosomal miRNA-21 plays a key role in hMSC paracrine-mediated enhancement of human engineered cardiac tissue contractile performance. In testing this hypothesis, I will directly address an NHLBI topic of special interest (HL-142) by studying the role of exosomes as paracrine mediators in cardiovascular disease. In Aim 1, I will identify the role of exosomal miRNA-21 in hMSC-mediated enhancement of hECT contractility by: 1) treating hECTs with exosomes derived from hMSCs with miRNA-21 inhibitor (Sub-aim 1A); and 2) formulated lipidoid nanoparticle delivery of miRNA-21 mimic into hECTs (Sub-aim 1B). In Aim 2, I will test the role of hMSC exosomes on recovery of contractility using in vitro hECT models of acquired (Sub-aim 2A) and genetic (Sub-aim 2B) non- ischemic heart failure. Overall, the project is designed to frame the research within a clinical context, and provide a rigorous multi- disciplinary training in tissue engineering, systems biology, electrophysiology, stem cell biology, and biochemistry as a solid foundation on which to build my career as a future physician-scientist.

项目摘要 非缺血性心肌病的新兴疗法涉及人类间充质茎的递送 细胞（HMSC）。临床试验文件对心脏收缩性的适度益处，强调了更好的需求 了解并利用有关HMSCS-毛肌细胞（HCMS）Interactome的基本机制。 关于HMSC介导的心脏疗法的最新研究表明，旁分泌信号传导-VIA分泌 因素 - 是降低心脏纤维化和增强血管生成的关键介体。此外，HMSC 旁分泌因子已显示通过改变心肌细胞通道/泵活性来影响收缩力。 但是，这些发现无法识别HMSC分泌组的关键组成部分，以增强收缩力。 我们建议利用三维人类工程心脏组织（HECT）作为体外模型 研究HMSC外泌体在增强心脏收缩力中的作用。 我们的实验室最近发现，HMSCS主要通过旁分泌增强了收缩力 信号传导，抵消HMSC-HCM杂细胞耦合的不良风险。重要的是，我们发现 HMSC分泌组的外泌体成分是必需的，足以介导HMSC-核酸 增强收缩力的增强。此外，通过使用系统生物学方法并整合Hect 收缩功能通过外泌体miRNA数据结果，我们预测外泌体miRNA-21作为铅候选者 负责HMSC旁分泌信号传导的收缩效应。我们后来用qpcr验证了 补充HMSC外泌体和HMSC总条件的miRNA-21水平增加 媒体相对于控制，激发了我们的中心假设，即外泌体miRNA-21在HMSC中起关键作用 旁分泌介导的人类工程性心脏组织收缩性能的增强。 在检验该假设时，我将通过研究该假设直接解决一个特殊关注的NHLBI主题（HL-142） 外泌体作为心血管疾病中旁分泌介质的作用。在AIM 1中，我将确定 HMSC介导的HECT收缩力增强中的外泌体miRNA-21通过：1）用 具有miRNA-21抑制剂（sub-aim 1a）的HMSC衍生的外泌体； 2）配制的脂肪纳米颗粒 将miRNA-21模拟于小公顷（子1B）传递。在AIM 2中，我将测试HMSC外泌体的作用 使用获得的（sub-aim 2a）和遗传（sub-aim 2b）的非体外HECT模型恢复收缩力 缺血性心力衰竭。 总体而言，该项目旨在在临床背景下进行研究，并提供严格的多种多样 组织工程，系统生物学，电生理学，干细胞生物学和 生物化学是建立我作为未来身体科学家的职业的坚实基础。