Characterization of the Cardiac Progenitor Cell Exosomes for Optimal Therapeutics

摘要 干细胞疗法在恢复心脏功能和促进组织再生方面的潜力 已完成的和正在招募的临床试验证明了这一点。然而，这些临床上的有益效果 使用成人CP的试验(是适度的。我们已经证明新生儿CPC在治疗中具有优越的疗效 修复受损的心脏与aCP相比，最近发现ncks的有益作用是 通过分泌体的旁分泌机制来调节。来自不同类型茎的外体 细胞/祖细胞已被证明介导干细胞触发的对受损心脏的治疗作用 通过他们的miRNA货物。因此，我们假设nCPC中的HSF1促进外切体的产生， 功能性外体miRNAs货物，以及对受体细胞的外体获取。.的存在 NCPC外切体中包括miR199a、miR590和miR146a在内的治疗性miRNAs刺激 靶向特定基因诱导的心肌细胞增殖和抑制细胞凋亡和纤维化 以恢复受伤心脏的心功能。为了验证我们的假设，我们提出了三个具体的 目标。目的1阐明HSF1在外体生物发生和外体运输中的功能作用 监管。我们将研究hsf1在ncpc中的敲除是否会取消它们优越的生成能力。 功能EXO和EXO货物，如果HSF1在aCRC中过表达将达到类似的能力 核祖细胞在EXO治疗功能中的作用。目标2将确定主要的外切体分子靶标 并确定HSF1是否是获得外切体所必需的。我们 将确定外切体的细胞靶点，以及HSF1是否对供体EXOS的保留是必要的。我们将研究 Hmer-1、Clic5、TRAF6和IRAK1是否为心功能恢复的主要分子靶点。目标3 将确定治疗性miRNAs是否介导外切体的效应，以及是否进一步 MiRNA浓缩可实现最佳的心脏恢复。我们将研究外体miRs-199a的影响， 590、146a对心脏恢复的影响。

SUMMARY The potential of stem cell therapies to restore heart function and promote tissue regeneration in response to injury is evident by the completed and recruiting clinical trials. However, the beneficial effects in these clinical trials using adult CPCs ( are modest. We have demonstrated that neonatal CPCs have superior efficacy in repairing the injured heart compared to aCPCs and recently, revealed that the nCPCs beneficial effect is mediated by a paracrine mechanism through a secretome. Exosomes derived from various type of stem cells/progenitor cells have been shown to mediate stem cell-triggered therapeutic effects on the injured heart through their miRNA cargo. Thus, we hypothesize that HSF1 in nCPCs promotes production of exosomes, functional exosomal miRNAs cargo, and exosome acquisition to recipient cells. The presence of therapeutic miRNAs including miR199a, miR590 and miR146a in nCPC exosomes stimulates cardiomyocyte proliferation and suppresses apoptosis and fibrosis by targeting specific genes leading to the restoration of cardiac function in the injured heart. To test our hypothesis, we proposed three specific aims. Aim 1 will elucidate the functional role of HSF1 in exosome biogenesis and exosomal cargo regulation. We will examine whether HSF1 knockdown in nCPCs will abolish their superior ability in generating functional EXO and EXO cargos, and if HSF1 overexpression in aCPCs will achieve comparable capability to that of nCPCs in EXO therapeutic functionality. Aim 2 will identify the major exosome molecular target in exosome-recipient cardiac cells and determine whether HSF1 is essential for exosome acquisition. We will identify cellular targets of exosomes and if HSF1 is essential for the retention of donor EXOs. We will examine whether Homer1, Clic5, TRAF6 and IRAK1 as EXO major molecular targets in cardiac function recovery. Aim 3 will determine whether the therapeutic miRNAs mediate the effect of exosomes and whether further miRNA enrichment achieves optimal cardiac recovery. We will examine the effects of exosomal miRs-199a, 590, 146a on cardiac recovery.