Cardiosphere-derived cells (CDCs) exert strong disease-modifying activity in various models of heart failure. In the original funding period of this R01, we tested, and ended up supporting, the hypothesis that the therapeutic benefits of CDCs are mediated by exosomes (nanosized lipid bilayer vesicles that are enriched in noncoding RNAs). Indeed, the concepts first articulated here for heart-derived cells have since been generalized to virtually all other cell types under development as therapeutic candidates for heart failure; even pluripotent stem cell products are now acknowledged to act largely via exosome secretion. We and others have also established that many, if not most, of the effects of exosomes are mediated by their RNA contents, specifically miRs and other noncoding RNAs (ncRNAs). The emerging paradigm is as follows: CDCs secrete exosomes which transfer payloads into target cells, inducing transcriptomic and phenotypic changes that underlie the benefits of CDC therapy. As a corollary of our work to define mechanisms of action of cardiac cell therapy, we have come to realize that exosomes are themselves viable next-generation therapeutic candidates. Potential advantages over the parent cells include product stability, immune tolerability, and the ability to achieve efficacy with simple intravenous (IV) administration. The major focus of the first funding period was on unaltered exosomes produced by primary wild type CDCs. In this competitive renewal, we will manipulate exosomes both by alterations of the parent cell and directly, after isolation. Such manipulation can increase potency as well as improve targeting. Technological and conceptual advances now make it possible to envision a novel, stable, cell-free therapeutic agent that can effectively target heart failure when delivered IV. Here we propose to develop trenchant methods to assess exosome biodistribution, to enhance exosome targeting and potency, and to immortalize CDCs as a stable source of therapeutic exosomes.
The specific aims are designed to answer four inter-connected questions: What is the biodistribution of exosomes when delivered IV (Aim 1)? Can we facilitate tissue/cell-directed targeting of exosomes when delivered IV (Aim 2)? Can we immortalize CDCs to generate therapeutically potent and consistent exosomes (Aim 3)? Combining the insights from these various approaches, can we create and select a therapeutic candidate of optimal efficacy for modifying heart failure in vivo after IV administration (Aim 4)? We will create useful models (fate-mapping mTmG mice) and methods (exosomal Cre loading, targeting strategies, CDC immortalization) which will advance our mechanistic understanding of exosome biology. Meanwhile, the proposed work constitutes an important translational step towards the development of exosomes as off-the- shelf therapeutic candidates. CDCs are already in advanced clinical testing, but living cells have limitations relative to cell-free products. Thus, our proposal, focusing on genetically-enhanced exosomes (as cell-free derivatives of immortalized CDCs), opens up new treatment options for heart failure.

Public Health Relevance

Cardiosphere-derived cells (CDCs), which are effective in various models of heart failure, work via the secretion of exosomes. Here we propose to develop trenchant methods to assess exosome biodistribution, to enhance exosome targeting and potency, and to immortalize CDCs as a stable source of therapeutic exosomes. The studies will advance our mechanistic understanding of exosome biology, while constituting an important translational step towards the development of exosomes as off-the-shelf cell-free agents to treat heart failure.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL124074-07
Application #
9880442
Study Section
Cardiac Contractility, Hypertrophy, and Failure Study Section (CCHF)
Program Officer
Wong, Renee P
Project Start
2014-07-11
Project End
2022-01-31
Budget Start
2020-02-01
Budget End
2021-01-31
Support Year
7
Fiscal Year
2020
Total Cost
Indirect Cost
Name
Cedars-Sinai Medical Center
Department
Type
DUNS #
075307785
City
Los Angeles
State
CA
Country
United States
Zip Code
90048
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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
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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
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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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