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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要一种治疗非缺血性心肌病的新兴疗法涉及人类间充质干细胞的输送细胞（hMSC）的临床试验证明对心肌收缩力有一定的益处，强调需要更好的方法。了解和利用控制 hMSCs-心肌细胞 (hCM) 相互作用的潜在机制。最近对 hMSC 介导的心脏治疗的研究表明，旁分泌信号传导——通过分泌因子——是减少心脏纤维化和增强血管生成的重要介质。旁分泌因子已被证明通过改变心肌细胞离子通道/泵活性来影响收缩力。然而，这些发现未能确定 hMSC 分泌蛋白组增强收缩力的关键成分。我们建议利用三维人体工程心脏组织（hECT）作为体外模型研究 hMSC 外泌体在增强心肌收缩力中的作用。我们实验室最近发现 hMSC 主要通过旁分泌增强 hECT 收缩力重要的是，我们发现，hMSC-hCM 异细胞偶联的不利风险。 hMSC 分泌组的外泌体成分对于 hMSC 旁分泌介导的作用是必要且充分的此外，通过利用系统生物学方法并整合 hECT 来增强 hECT 收缩性。根据外泌体 miRNA 数据的收缩功能结果，我们预测外泌体 miRNA-21 作为主要候选者我们后来通过 qPCR 验证了这一点。补充 hMSC 外泌体和 hMSC 总条件化的 hECT 中 miRNA-21 水平增加培养基相对于对照，激发了我们的中心假设：外泌体 miRNA-21 在 hMSC 中发挥关键作用旁分泌介导的人类工程心脏组织收缩性能的增强。在检验这一假设时，我将通过研究以下内容直接解决 NHLBI 特别感兴趣的主题 (HL-142) 外泌体作为旁分泌介质在心血管疾病中的作用。在目标 1 中，我将确定外泌体的作用。外泌体 miRNA-21 在 hMSC 介导的 hECT 收缩性增强中的作用：1）用源自 hMSC 的外泌体与 miRNA-21 抑制剂（子目标 1A）和 2) 配制的类脂质纳米颗粒；将 miRNA-21 模拟物递送至 hECT（子目标 1B）在目标 2 中，我将测试 hMSC 外泌体的作用。使用获得性（子目标 2A）和遗传性（子目标 2B）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框架研究，并提供严格的多方面研究组织工程、系统生物学、电生理学、干细胞生物学等学科培训生物化学为我未来成为一名医生科学家的职业生涯奠定了坚实的基础。