Exosome-mediated cardioprotection and regeneration

外泌体介导的心脏保护和再生

• 批准号: 8759304
• 负责人: EDUARDO MARBAN
• 金额: $ 42.5万
• 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/8759304
• 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; 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; public health relevance; 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（由申请人提供）：心肌衍生细胞（CDC）可有效减少心肌梗塞（MI）后患者的疤痕大小和再生可行的心肌，并在MI动物模型中发挥急性心脏保护作用。 CDC的主要好处似乎是由间接机制介导的。在这里，我们检验了总体假设，即CDC的间接作用是由外泌体介导的。外泌体分泌多种细胞类型，是30-100 nm脂质双层囊泡，富含包括microRNA（miRS）在内的RNA中。我们的初步数据表明，在小鼠急性MI模型中，人类疾病节分泌的外泌体再现了CDC的治疗益处，对于CDC的治疗益处是必不可少的。将CDC外诊断到受伤的心脏中，模仿CDC移植的结构和功能益处；相反，CDC对外泌体分泌的抑制作用消除了移植的CDC的治疗益处。并非所有的外泌体都是有益的：从真皮成纤维细胞中注射外泌体，无法改善MI后结局的对照细胞，没有治疗益处。 CDC - 异位体减少了急性心肌细胞死亡和炎症细胞因子释放，同时减少了MI后心脏的LV重塑和纤维化。来自microRNA阵列的初步数据揭示了几个“签名miR”，这些“签名miR”在CDC-外泌体中高度上调。相比之下，质谱法表明CDC-诊断的蛋白质组成是常规的，并且与成纤维细胞外粘体相当。因此，我们假设：i）CDC-诊断物包含独特的miR互补，共同介导了CDC的许多或全部治疗作用； ii）CDC-外诊断及其成分miR有利地调节MI后心脏中的细胞凋亡，炎症和纤维化； iii）CDC - 外观改善功能恢复并增加了MI后的组织生存力； iv）CDC - 外观代表了心脏修复的可行“无细胞”治疗候选者。在这里，我们建议对CDC-诊断的完整RNA含量进行测序，以确定CDC-诊断的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-诊断作为治疗MI和HF的生物学产品。由于外泌体与可以主动朝目标迁移的活细胞不同，因此我们还试图测试新型的递送方法，以选择性地靶向CDC - 异肌细胞对受伤的肌细胞。该提案既是假设驱动的，又以产品为导向的。我们试图依靠最先进的科学方法和良好的临床前疾病模型来详细回答机械和翻译问题。 CDC（自体和同种异体性）已经在人类试验中，但是细胞作为治疗剂的局限性。因此，我们的建议重点是CDC-诊断（作为CDC的无细胞衍生物），具有潜在的转化价值。结果还将为急性MI后细胞介导的心脏保护和心脏再生的基本机制提供新的见解。