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: 9592986
• 负责人: Joshua Mayourian
• 金额: $ 3.89万
• 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 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9592986
• 关键词: Activities of Daily Living; Address; Apoptotic; Biochemistry; Biological Assay; Ca(2+)-Transporting ATPase; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cardiovascular Diseases; Cells; Clinical; Clinical Trials; Coupling; Data; Dilated Cardiomyopathy; Dimensions; Electrophysiology (science); Endoplasmic Reticulum; Fibrosis; Foundations; Future; Gene Expression; Genetic; Genetic Models; Heart; Heart failure; Human; Human Engineering; In Vitro; Ion Channel; L-Type Calcium Channels; 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; Training; Transplantation; United States National Institutes of Health; angiogenesis; cardiac tissue engineering; career; coronary fibrosis; design; exosome; human stem cells; immunoregulation; improved; in vitro Model; inhibitor/antagonist; innovation; insight; interest; lead candidate; mRNA Expression; mathematical model; multidisciplinary; mutant; nanoparticle delivery; 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.

项目总结 一种新的治疗非缺血性心肌病的方法是移植人间充质干细胞 细胞(HMSCs)。临床试验记录了对心脏收缩能力的适度好处，强调了更好地 了解并开发调控hMSCs-心肌细胞(HCMS)相互作用组的潜在机制。 最近对hMSC介导的心脏治疗的研究表明，旁分泌信号途径分泌 因子-是减少心脏纤维化和增强血管生成的关键介体。此外，hMSC 已有研究表明，旁分泌因子通过改变心肌细胞离子通道/泵的活性来影响收缩能力。 然而，这些发现未能确定hMSC分泌组中增强收缩能力的关键成分。 我们建议利用三维人类工程心脏组织(HECTs)作为体外模型 探讨hMSC外切体在增强心肌收缩能力中的作用。 我们的实验室最近发现，hMSCs主要通过旁分泌增强Hect的收缩能力。 信号转导，对抗hMSC-HCM异质细胞偶联的不利风险。重要的是，我们发现 HMSC分泌体的外体成分是hMSC旁分泌介导的必要条件和充分条件。 增强Hect的收缩能力。此外，通过利用系统生物学方法和将Hect 收缩功能结果与外体miRNA数据，我们预测外体miRNA-21是一个领先的候选 负责hMSC旁分泌信号的良好收缩效应。我们后来用qPCR验证了 补充hMSC外切体和hMSC总条件的hECT中miRNA-21水平升高 相对于对照的媒体，激励了我们的中心假设，即外体miRNA-21在hMSC中发挥关键作用 旁分泌介导的人工程化心脏组织收缩性能的增强。 为了验证这一假设，我将直接讨论NHLBI的一个特别感兴趣的主题(HL-142)，通过研究 外切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 外体miRNA-21在hMSC介导的hECT收缩增强中的作用：1)用 含miRNA-21抑制剂的人骨髓间充质干细胞来源的外体(亚目标1A)；和2)配制的类脂纳米粒 将miRNA-21模拟载体导入hECT(亚靶1B)。在目标2中，我将测试hMSC exosome在 利用获得性(亚目标2A)和遗传性(亚目标2B)非心肌细胞体外Hect模型恢复心肌收缩功能 缺血性心力衰竭。 总体而言，该项目旨在将研究框架置于临床背景下，并提供严格的多学科 组织工程、系统生物学、电生理学、干细胞生物学和 生物化学是我作为一名未来内科科学家的职业生涯的坚实基础。