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.
Cardiosphere-derived cells (CDCs) are effective in protecting against ischemic damage and regenerating viable myocardium in acute myocardial infarction, via indirect mechanisms. Here we test the overarching hypothesis that the beneficial effects of CDCs are mediated by nanosized lipid bilayer vesicles (exosomes) that are secreted by CDCs and taken up by neighboring heart cells. The proposed work will advance our mechanistic understanding of cell-mediated cardioprotection and regeneration, while constituting an important first step towards the development of CDC-derived exosomes as a novel cell-free therapeutic agent.
|Aminzadeh, Mark A; Rogers, Russell G; Fournier, Mario et al. (2018) Exosome-Mediated Benefits of Cell Therapy in Mouse and Human Models of Duchenne Muscular Dystrophy. Stem Cell Reports 10:942-955|
|Marbán, Eduardo; de Couto, Geoffrey (2018) Response by Marbán and de Couto to Letter Regarding Article, ""Exosomal MicroRNA Transfer Into Macrophages Mediates Cellular Postconditioning"". Circulation 137:212|
|Antes, Travis J; Middleton, Ryan C; Luther, Kristin M et al. (2018) Targeting extracellular vesicles to injured tissue using membrane cloaking and surface display. J Nanobiotechnology 16:61|
|Middleton, Ryan C; Rogers, Russell G; De Couto, Geoffrey et al. (2018) Newt cells secrete extracellular vesicles with therapeutic bioactivity in mammalian cardiomyocytes. J Extracell Vesicles 7:1456888|
|Cho, Jae Hyung; Zhang, Rui; Aynaszyan, Stephan et al. (2018) Ventricular Arrhythmias Underlie Sudden Death in Rats With Heart Failure and Preserved Ejection Fraction. Circ Arrhythm Electrophysiol 11:e006452|
|Cambier, Linda; Giani, Jorge F; Liu, Weixin et al. (2018) Angiotensin II-Induced End-Organ Damage in Mice Is Attenuated by Human Exosomes and by an Exosomal Y RNA Fragment. Hypertension 72:370-380|
|Marbán, Eduardo (2018) The Secret Life of Exosomes: What Bees Can Teach Us About Next-Generation Therapeutics. J Am Coll Cardiol 71:193-200|
|de Couto, Geoffrey; Gallet, Romain; Cambier, Linda et al. (2017) Exosomal MicroRNA Transfer Into Macrophages Mediates Cellular Postconditioning. Circulation 136:200-214|
|Eschenhagen, Thomas; Bolli, Roberto; Braun, Thomas et al. (2017) Cardiomyocyte Regeneration: A Consensus Statement. Circulation 136:680-686|
|Cambier, Linda; de Couto, Geoffrey; Ibrahim, Ahmed et al. (2017) Y RNA fragment in extracellular vesicles confers cardioprotection via modulation of IL-10 expression and secretion. EMBO Mol Med 9:337-352|
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