Exosome-mediated cardioprotection and regeneration

外泌体介导的心脏保护和再生

• 批准号: 8890879
• 负责人: EDUARDO MARBAN
• 金额: $ 41.86万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-11 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8890879
• 关键词: Accounting; Acute; Acute myocardial infarction; Allogenic; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Apoptosis; Apoptotic; Attenuated; Autologous; Biological Process; Biological Products; Cardiac Myocytes; Cell Therapy; Cell Transplantation; Cell physiology; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Cicatrix; Complement; Data; Dermal; Development; Disease model; Engineering; Fibroblasts; Fibrosis; Goals; Health; Heart; Human; Image; Inflammation; Inflammatory; Injection of therapeutic agent; Intravenous; Knock-out; Left Ventricular Remodeling; Life; Lipid Bilayers; Mass Spectrum Analysis; Mediating; Methods; MicroRNAs; Modeling; Molecular; Mus; Muscle Cells; Myocardial Infarction; Myocardium; Myosin Light Chains; Natural regeneration; Neoplasm Metastasis; Outcome; Pathway interactions; Patients; Proteins; RNA; Recovery of Function; Relative (related person); Resolution; Stem cells; Surface; Testing; Therapeutic; Therapeutic Agents; Therapeutic Effect; Tissue Viability; Transgenic Mice; Transplantation; Vesicle; Work; angiogenesis; base; cardiac regeneration; cardiac repair; cell type; cytokine; heart cell; improved; in vitro activity; in vivo; injured; insight; nanosized; novel; pre-clinical; relating to nervous system; targeted delivery; tissue regeneration; trafficking; tumor

DESCRIPTION (provided by applicant): Cardiosphere-derived cells (CDCs) are effective in reducing scar size and regenerating viable myocardium in patients after myocardial infarction (MI), and exert acute cardioprotective effects in animal models of MI. The major benefits of CDCs appear to be mediated by indirect mechanisms. Here we test the overarching hypothesis that the indirect effects of CDCs are mediated by exosomes. Secreted by a wide range of cell types, exosomes are 30-100 nm lipid bilayer vesicles that are enriched in RNAs including microRNAs (miRs). Our preliminary data indicate that exosomes secreted by human CDCs reproduce the therapeutic benefits of CDCs, and are indispensable for the therapeutic benefits of CDCs, in a mouse acute MI model. Injection of CDC-exosomes into the injured heart mimics the structural and functional benefits of CDC transplantation; conversely, inhibition of exosome secretion by CDCs abrogates the therapeutic benefits of transplanted CDCs. Not all exosomes are salutary: Injection of exosomes from dermal fibroblasts, control cells which do not improve post-MI outcomes, had no therapeutic benefit. CDC-exosomes decreased acute cardiomyocyte death and inflammatory cytokine release, while attenuating LV remodeling and fibrosis in the post-MI heart. Preliminary data from microRNA arrays reveal several "signature miRs" that are highly up-regulated in CDC- exosomes. In contrast, mass spectrometry indicates that the protein composition of CDC-exosomes is conventional and comparable to that of fibroblast-exosomes. Thus, we hypothesize that: i) CDC-exosomes contain a unique complement of miRs that, collectively, mediate many, or all, of the therapeutic effects of CDCs; ii) CDC-exosomes and their constituent miRs favorably modulate apoptosis, inflammation and fibrosis in the post-MI heart; iii) CDC-exosomes improve functional recovery and increase tissue viability post-MI; iv) CDC-exosomes represent a viable "cell free" therapeutic candidate for cardiac repair. Here we propose to sequence the full RNA content of CDC-exosomes, to determine the molecular basis underlying the therapeutic benefit of CDC-exosomes, and to optimize delivery strategies in vivo. Our ultimate goal is to develop CDC-exosomes as a biologic product to treat MI and HF. Because exosomes are unlike live cells that can migrate actively towards a target, we also seek to test novel delivery methods to target CDC-exosomes selectively to injured myocytes. The proposal is both hypothesis-driven and product-oriented. We seek to answer both mechanistic and translational questions in detail, relying upon state-of-the-art scientific methods and well-established preclinical disease models. CDCs (both autologous and allogeneic) are already in human trials, but cells have their limitations as therapeutic agents. Thus, our proposal, focusing on CDC-exosomes (as cell-free derivatives of CDCs), is of potentially significant translational value. The results will also open up novel insights into the fundamental mechanisms of cell-mediated cardioprotection and cardiac regeneration after acute MI.

描述(申请人提供)：心球来源的细胞(CDCs)在减少心肌梗死(MI)后患者的疤痕大小和再生存活心肌方面有效，并在MI的动物模型中发挥急性心脏保护作用。疾病预防控制中心的主要好处似乎是通过间接机制实现的。在这里，我们测试了CDC的间接效应是由外切体介导的最重要的假设。Exosome由多种细胞类型分泌，是30-100 nm的脂双层囊泡，富含包括microRNAs(MiRs)在内的RNA。我们的初步数据表明，在小鼠急性心肌梗死模型中，人CDCs分泌的外切体复制了CDCs的治疗益处，并且对于CDCs的治疗益处是不可或缺的。将CDC外切体注射到受损心脏可模拟CDC移植的结构和功能益处；相反，抑制CDC外切体的分泌则剥夺了移植的CDC的治疗益处。并不是所有的Exosome都是有益的：注射来自真皮成纤维细胞的Exosome，作为对照细胞，不能改善MI后的结果，没有治疗益处。CDC-exosome可减少急性心肌细胞死亡和炎性细胞因子的释放，同时减轻心肌梗死后心脏的左室重构和纤维化。来自microRNA阵列的初步数据显示，CDC-exosome中有几个高度上调的“标志性miRs”。相反，质谱仪表明CDC外切体的蛋白质组成是常规的，与成纤维细胞外切体的蛋白质组成相当。因此，我们假设：i)CDC-exosome包含一组独特的miRs，它们共同介导CDC的许多或全部治疗效果；ii)CDC-exosome及其组成的miRs可有利地调节心肌梗死后的细胞凋亡、炎症和纤维化；iii)CDC-exosome可改善心肌梗死后的功能恢复并增加组织存活；iv)CDC-exosome是一种可行的“无细胞”心脏修复候选药物。在这里，我们建议对CDC-exosome的完整RNA内容进行测序，以确定CDC-exosome治疗益处的分子基础，并优化体内的递送策略。我们的最终目标是开发CDC外切体作为治疗心肌梗死和心力衰竭的生物产品。由于外切体不同于可以主动向靶点迁移的活细胞，我们还寻求测试新的递送方法，以选择性地将CDC-外切体靶向受损的心肌细胞。该提议既是假设驱动的，也是产品导向的。我们依靠最先进的科学方法和成熟的临床前疾病模型，寻求详细回答机械性和翻译性问题。CDCs(自体和同种异体)已经在人体试验中，但细胞作为治疗剂有其局限性。因此，我们的建议，重点是CDC-exosome(CDC-exosome)(CDC的无细胞衍生物)，具有潜在的重要翻译价值。这一结果还将为急性心肌梗死后细胞介导的心脏保护和心脏再生的基本机制开辟新的视角。