JACC CardioOncol. 2021 Sep;3(3):
428-440
Connor G O'Brien,
Mehmet Ozgun Ozen,
Gentaro Ikeda,
Evgeniya Vaskova,
Ji Hye Jung,
Nathan Bayardo,
Michelle Rai Santoso,
Liye Shi,
Christine Wahlquist,
Zewen Jiang,
Yunshin Jung,
Yitian Zeng,
Elizabeth Egan,
Robert Sinclair,
Adrian Gee,
Ronald Witteles,
Mark Mercola,
Katrin J Svensson,
Utkan Demirci,
Phillip C Yang.
Background: Anthracycline-induced cardiomyopathy (AIC) is a significant source of morbidity and mortality in cancer survivors. The role of mesenchymal stem cells (MSCs) in treating AIC was evaluated in the SENECA trial, a Phase 1 National Heart, Lung, and Blood Institute-sponsored study, but the mechanisms underpinning efficacy in human tissue need clarification.Objectives: The purpose of this study was to perform an in vitro clinical trial evaluating the efficacy and putative mechanisms of SENECA trial-specific MSCs in treating doxorubicin (DOX) injury, using patient-specific induced pluripotent stem cell-derived cardiomyocytes (iCMs) generated from SENECA patients.
Methods: Patient-specific iCMs were injured with 1 μmol/L DOX for 24 hours, treated with extracellular vesicles (EVs) from MSCs by either coculture or direct incubation and then assessed for viability and markers of improved cellular physiology. MSC-derived EVs were separated into large extracellular vesicles (L-EVs) (>200 nm) and small EVs (<220nm) using a novel filtration system.
Results: iCMs cocultured with MSCs in a transwell system demonstrated improved iCM viability and attenuated apoptosis. L-EVs but not small EVs recapitulated this therapeutic effect. L-EVs were found to be enriched in mitochondria, which were shown to be taken up by iCMs. iCMs treated with L-EVs demonstrated improved contractility, reactive oxygen species production, ATP production, and mitochondrial biogenesis. Inhibiting L-EV mitochondrial function with 1-methyl-4-phenylpyridinium attenuated efficacy.
Conclusions: L-EV-mediated mitochondrial transfer mitigates DOX injury in patient-specific iCMs. Although SENECA was not designed to test MSC efficacy, consistent tendencies toward a positive effect were observed across endpoints. Our results suggest a mechanism by which MSCs may improve cardiovascular performance in AIC independent of regeneration, which could inform future trial design evaluating the therapeutic potential of MSCs.
Keywords: AIC, anthracycline induced cardiomyopathy; DOX, doxorubicin; DZR, dexrazoxane; EV, extracellular vesicle; L-EV, large extracellular vesicle; MPP+, 1-methyl-4-phenylpyrindinium; MSC, mesenchymal stem cell; MSC-EV, mesenchymal stem cell derived extracellular vesicle; MTDR, MitoTracker Deep Red; MTG, MitoTracker Green; RBC, red blood cell; ROS, reactive oxygen species; S-EV, small extracellular vesicle; anthracycline; cardiomyopathy; heart failure; iCM, induced cardiomyocyte