Smarter exosomes derived from engineered MSCs promote neo-vascularization

源自工程化 MSC 的更智能的外泌体可促进新血管形成

• 批准号: 10078974
• 负责人: Min Liu
• 金额: $ 50.5万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-15 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10078974
• 关键词: Adverse effects; Angiogenesis Inhibitors; Animals; Caliber; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cause of Death; Cell Therapy; Cell membrane; Cells; Data; Dose; Effectiveness; Encapsulated; Endothelial Cells; Engineering; Extracellular Space; Family; Future; Goals; Heart; Immune; In Vitro; Injections; Intravenous; Liposomes; Mediating; Mesenchymal Stem Cells; Modality; Modeling; Morphogenesis; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardial tissue; Myocardium; Natural regeneration; Outcome; Parents; Pathway interactions; Patients; Physiological; Pilot Projects; Play; Pre-Clinical Model; Property; Proteins; Publishing; Recovery; Reporting; Risk; Role; Safety; Signal Pathway; Solid; Stress; THBS1 gene; Technology; Testing; Therapeutic Effect; Thrombospondin 1; Time; Transfection; Translational Research; Validation; Vascularization; Vesicle; angiogenesis; base; blood vessel development; cardiac regeneration; cardiac repair; cardiovascular health; clinical application; clinical implementation; compare effectiveness; effective therapy; exosome; experimental study; extracellular vesicles; genetic manipulation; genome editing; heart damage; heart function; heart preservation; improved; in vivo; injured; innovation; intravenous administration; intravenous injection; ischemic injury; loss of function; myocardial injury; nanoparticle; neovascularization; overexpression; paracrine; preconditioning; repaired; response; stem cell exosomes; stem cell survival; stem cell therapy; stem cells; success; therapy outcome; tissue regeneration; tissue repair; transcription factor; transdifferentiation; vector; vesicular release

Project Summary/Abstract Mesenchymal stem cell (MSC) therapy has shown tremendous promise for enabling heart tissue repair after ischemic injury. The therapeutic effects of stem cells are mediated by paracrine factors. Several studies illustrate that exosomes (EXO) derived from stem cells play a critical role in stem cell mediated therapy of ischemic myocardium via increasing angiogenesis. From the translational perspective, EXO have greater salutary therapeutic effects than whole cells: EXO have fewer potential adverse effects, less immune rejection, are more amenable to manipulation, and EXO cargo can be modified by preconditioning or genetic manipulation. GATA- 4, a cardiac transcription factor, promotes cardiac morphogenesis, extends cardiomyocyte (CM) survival and preserves cardiac function via regulating various bioactive molecules and activating cardiac protective miRs. Our published data indicate that MSC overexpressing GATA-4 (MSCGATA-4) increase MSC survival, protect native CM and promote angiogenesis, compared to vector-transfect MSC (MSCnull). EXO derived from EXO derived from GATA-4 overexpressing GATA-4 (ExoGATA-4) are more efficient than EXO from vector-transfected MSC (Exonull) in protecting CM from ischemic injury. In our pilot study, ExoGATA-4 are enriched with pro-angiogenic miRs, which regulate vascularization. ExoGATA-4 downregulate the expression of thrombospondin 1, a well-known endogenous inhibitor of neovascularization. The overarching goal of this project is to exploit effectiveness of GATA-4 conferred EXO than the ordinary EXO in repairable effects of ExoGATA-4. Our central hypothesis is that the ExoGATA-4 are enriched with pro-angiogenic miRs and proteins that are transferred into recipient cells and active multiple signaling pathways, leading to angiogenesis and cardiac repair. Three Specific Aims are proposed: Aim 1, to determine the role of miRs carried by EXO in response to GATA-4 transfection. Aim 2, to demonstrate if the transferred bioactive molecules play a critical role in ExoGATA-4 mediated angiogenesis. Aim 3, to test the hypothesis that EXO derived from GATA-4 overexpressing MSCs are more effective than that from vector-transfected MSCs in promoting angiogenesis and myocardial regeneration. The proposed studies are innovative because no previous study has systematically examined the effectiveness of ExoGATA-4, with particular focus on the efficacy of EXO mediated angiogenesis and ischemic heart repair following systemic administration. Intravenous injection offers the advantage of enabling repeated treatments without the stress of repeated intramyocardial injections. The proposed study is highly significant because it will evaluate the concept that the effectiveness of cellular therapy can be reproduced by cell-free EXO and will explore this new modality for future clinical application by intravenous administration. This project will be the first systematic effort aimed at developing a completely new strategy in use of EXO administration in lieu of cells, which would revolutionize cell therapy. The outcome will offer a transformative validation and guide translational research and putative therapies based on enhanced angiogenic properties of EXO from genome-edited MSCs. 1

项目概要/摘要 间充质干细胞 (MSC) 疗法在促进心脏组织修复方面显示出巨大的前景 缺血性损伤。干细胞的治疗作用是由旁分泌因子介导的。多项研究表明 来自干细胞的外泌体（EXO）在干细胞介导的缺血性治疗中发挥着关键作用 通过增加血管生成来增强心肌。从转化的角度来看，EXO有更大的价值 比全细胞治疗效果：EXO潜在不良反应更少，免疫排斥更少，更多 易于操作，并且 EXO 货物可以通过预处理或基因操作进行修改。加塔- 4，一种心脏转录因子，促进心脏形态发生，延长心肌细胞 (CM) 存活时间和 通过调节各种生物活性分子和激活心脏保护性 miR 来保护心脏功能。 我们发表的数据表明，MSC 过表达 GATA-4 (MSCGATA-4) 可增加 MSC 存活率，保护天然 与载体转染 MSC (MSCnull) 相比，CM 并促进血管生成。 EXO 衍生 EXO 衍生 来自 GATA-4 的过表达 GATA-4 (ExoGATA-4) 比来自载体转染的 MSC 的 EXO 更有效 (Exonull) 保护 CM 免受缺血性损伤。在我们的初步研究中，ExoGATA-4 富含促血管生成物质 miR，调节血管形成。 ExoGATA-4 下调血小板反应蛋白 1 的表达，血小板反应蛋白 1 是一种众所周知的 新血管形成的内源性抑制剂。该项目的总体目标是利用 GATA-4赋予EXO比普通EXO在ExoGATA-4的可修复效果上。我们的中心假设是 ExoGATA-4 富含促血管生成 miR 和蛋白质，可转移到受体细胞中，并且 激活多种信号通路，导致血管生成和心脏修复。三个具体目标是 建议：目标 1，确定 EXO 携带的 miR 在响应 GATA-4 转染中的作用。目标2， 证明转移的生物活性分子是否在 ExoGATA-4 介导的血管生成中发挥关键作用。目的 3，检验来自GATA-4过表达MSC的EXO比来自GATA-4过表达MSC的EXO更有效的假设 载体转染的间充质干细胞促进血管生成和心肌再生。拟议的研究是 之所以具有创新性，是因为之前没有研究系统地检验过 ExoGATA-4 的有效性， 特别关注全身性手术后 EXO 介导的血管生成和缺血性心脏修复的功效 行政。静脉注射的优点是可以重复治疗，而无需担心 重复心肌内注射。拟议的研究非常重要，因为它将评估 细胞疗法的有效性可以通过无细胞 EXO 重现的概念，并将探索这种新的 未来临床应用的方式是静脉注射。该项目将是第一个系统性的工作 旨在开发一种全新的策略，使用 EXO 管理代替细胞，这将 彻底改变细胞疗法。结果将提供变革性的验证并指导转化研究和 基于来自基因组编辑的 MSC 的 EXO 增强的血管生成特性的推定疗法。 1