Microvesicles from Adipose-derived Stem Cells for Ischemic Heart Repair

 描述（由申请人提供）：缺血性心脏病在全球范围内流行，迫切需要创新的治疗方法，尽管这些疾病的医学、介入和手术治疗取得了重大进展。基于微泡（MV）的治疗性血管生成是新兴的缺血性疾病，因为肿瘤的干细胞方法的关注。MV是从细胞释放的亚微米颗粒，包含蛋白质、mRNA、microRNA和生物活性脂质的包。此外，MV最近被认为是细胞间通讯的信号传递载体。在我们目前的试点资助期间，我们已经证明，来自脂肪源性干细胞（ASC），特别是来自内皮分化培养基（EDM）预处理的ASC的MV，在体外，离体和体内（基质胶塞）促进血管生成。促血管生成的潜在机制可能是通过MV将microRNA-31从ASC递送至血管内皮细胞，其中FIH 1（抑制HIF-1的因子）被靶向和抑制。在本申请中，我们的中心假设是移植ASC衍生的MV促进缺血性心脏心肌梗死中的血管生成。我们将通过更广泛的努力来补充这种方法，以继续为我们的假设驱动的机械实验管道提供支持。目的1是确定从EDM预处理的ASCs释放的MV在促进缺血心脏中的血管生成中的作用。将在严重联合免疫缺陷（NOD SCID）小鼠心肌梗死（MI）后给予MV。随后将评价心脏功能、细胞增殖和凋亡、纤维化面积和毛细血管密度。目的2探讨miR-31信号通路在微血管细胞间通讯中的作用。我们将探讨缺氧诱导因子-1（HIF-1）的活性，这已被报道被FIH1抑制，并将扩大我们对miR-31/FIH1/HIF-1通路在MV移植缺血心肌中的作用的理解。目的3是确定ASC衍生的MV中miR-31对MI后心脏修复的作用。将研究MV给药的不同剂量、时间和给药途径对MI后心脏修复的影响。为了增加我们研究的通量，将在EDM预处理的MV供体ASC中或直接在MV中操作miR-31。在这项研究计划的结论中，我们将表征miR-31/FIH1/HIF 1通路在缺血性心脏病的基于MV的治疗中的作用。通过MV递送血管生成miRNA来调节血管内皮细胞中的miRNA组成可能是治疗缺血性心脏病的创新策略。

 DESCRIPTION (provided by applicant): The worldwide epidemic of ischemic heart diseases urgently requires innovative treatments in spite of the significant advances in medical, interventional and surgical therapy for these diseases. A microvesicle (MV)-based therapeutic angiogenesis is emerging for ischemic diseases because of tumorigenic concern of the stem cell approach. MVs are submicron particles that are released from cells, and contain a package of protein, mRNA, microRNA and bioactive lipid. In addition, MV has recently been recognized as a signaling transferring vehicle in intercellular communication. In our current pilot funding period, we have demonstrated that MVs from adipose-derived stem cells (ASCs), particularly from endothelial differentiation medium (EDM)-preconditioned ASCs, promote angiogenesis in vitro, ex vivo and in vivo (matrigel plugs). An underlying mechanism of the proangiogenesis may be the delivery of microRNA-31 via MVs from ASCs to vascular endothelial cells in which FIH1 (factor inhibiting HIF-1) is targeted and suppressed. In this application, our central hypothesis i that the transplantation of ASC-derived MVs promotes angiogenesis in ischemic heart myocardial infarction. We will complement this approach with broader efforts to continue to feed our pipeline of hypothesis-driven mechanistic experiments. Aim 1 is to determine the role of MVs released from EDM-preconditioned ASCs in promoting angiogenesis in the ischemic heart. The MVs will be administered after myocardial infarction (MI) in severe combined immunodeficiency (NOD SCID) mice. The cardiac function, cell proliferation and apoptosis, fibrotic area, and capillary density will subsequently be evaluated. Aim 2 is to investigate the role of miR-31 signaling in intercellular communication through the MVs. We will explore the activity of hypoxia inducible factor-1 (HIF-1), which has been reported to be inhibited by FIH1, and will broaden our understanding of the role of miR-31/FIH1/HIF-1 pathway in MV-transplanted ischemic myocardium. Aim 3 is to determine the effect of miR-31 in ASC-derived MVs on cardiac repair after MI. The effects of different dosing, timing and delivery routes of administration of MVs on cardiac repair after MI will be investigated. To increase the throughput of our investigation, miR-31 will be manipulated in EDM-preconditioned MV-donor ASCs or in MVs directly. At the conclusion of this research proposal we will have characterized the role of miR-31/FIH1/HIF1 pathway in MV- based therapy for ischemic heart diseases. Modulation of miRNA composition in vascular endothelial cells by delivering angiogenic miRNAs via MV may be an innovative strategy for the treatment of ischemic heart diseases.