Cardiac recovery via extended cell-free delivery of extracellular vesicles secreted by cardiomyocytes derived from induced pluripotent stem cells

  • Nature Biomedical Engineeringvolume 2pages293303 (2018)
  • doi:10.1038/s41551-018-0229-7
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The ability of extracellular vesicles (EVs) to regulate a broad range of cellular processes has recently been exploited for the treatment of diseases. For example, EVs secreted by therapeutic cells injected into infarcted hearts can induce recovery through the delivery of cell-specific microRNAs. However, retention of the EVs and the therapeutic effects are short-lived. Here, we show that an engineered hydrogel patch capable of slowly releasing EVs secreted from cardiomyocytes (CMs) derived from induced pluripotent stem cells reduced arrhythmic burden, promoted ejection-fraction recovery, decreased CM apoptosis 24 h after infarction, and reduced infarct size and cell hypertrophy 4 weeks post-infarction when implanted onto infarcted rat hearts. We also show that EVs are enriched with cardiac-specific microRNAs known to modulate CM-specific processes. The extended delivery of EVs secreted from induced-pluripotent-stem-cell-derived CMs into the heart may help us to treat heart injury and to understand heart recovery.

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  1. 1.

    Stastna, M. & Van Eyk, J. E. Investigating the secretome: lessons about the cells that comprise the heart. Circ. Cardiovasc. Genet. 5, o8–o18 (2012).

  2. 2.

    Wei, K. et al. Epicardial FSTL1 reconstitution regenerates the adult mammalian heart. Nature 525, 479–485 (2015).

  3. 3.

    Ohno, S. et al. Systemically injected exosomes targeted to EGFR deliver antitumor microRNA to breast cancer cells. Mol. Ther. 21, 185–191 (2013).

  4. 4.

    Raposo, G. & Stoorvogel, W. Extracellular vesicles: exosomes, microvesicles, and friends. J. Cell Biol. 200, 373–383 (2013).

  5. 5.

    Tkach, M. & Théry, C. Communication by extracellular vesicles: where we are and where we need to go. Cell 164, 1226–1232 (2016).

  6. 6.

    Lo Cicero, A., Stahl, P. D. & Raposo, G. Extracellular vesicles shuffling intercellular messages: for good or for bad. Curr. Opin. Cell Biol. 35, 69–77 (2015).

  7. 7.

    Emanueli, C., Shearn, A. I. U., Angelini, G. D. & Sahoo, S. Exosomes and exosomal miRNAs in cardiovascular protection and repair. Vasc. Pharmacol. 71, 24–30 (2015).

  8. 8.

    Stoorvogel, W. Functional transfer of microRNA by exosomes. Blood 119, 646–648 (2012).

  9. 9.

    Olson, E. N. MicroRNAs as therapeutic targets and biomarkers of cardiovascular disease. Sci. Transl. Med. 6, 239ps3 (2014).

  10. 10.

    Malik, Z. A. et al. Cardiac myocyte exosomes: stability, HSP60, and proteomics. AJP Hear. Circ. Physiol. 304, H954–H965 (2013).

  11. 11.

    Stamm, C. et al. Autologous bone-marrow stem-cell transplantation for myocardial regeneration. Lancet 361, 45–46 (2003).

  12. 12.

    Garbern, J. C. & Lee, R. T. Cardiac stem cell therapy and the promise of heart regeneration. Cell Stem Cell 12, 689–698 (2013).

  13. 13.

    Segers, V. F. M. & Lee, R. T. Stem-cell therapy for cardiac disease. Nature 451, 937–942 (2008).

  14. 14.

    Toma, C., Pittenger, M. F., Cahill, K. S., Byrne, B. J. & Kessler, P. D. Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation 105, 93–98 (2002).

  15. 15.

    Vrtovec, B. et al. Effects of intracoronary CD34+ stem cell transplantation in nonischemic dilated cardiomyopathy patients: 5-year follow-up. Circ. Res. 112, 165–173 (2013).

  16. 16.

    Karantalis, V. & Hare, J. M. Use of mesenchymal stem cells for therapy of cardiac disease. Circ. Res. 116, 1413–1430 (2015).

  17. 17.

    Godier-Furnémont, A. F. G. et al. Composite scaffold provides a cell delivery platform for cardiovascular repair. Proc. Natl Acad. Sci. USA 108, 7974–7979 (2011).

  18. 18.

    Gallet, R. et al. Exosomes secreted by cardiosphere-derived cells reduce scarring, attenuate adverse remodelling, and improve function in acute and chronic porcine myocardial infarction. Eur. Heart J. 38, 201–211 (2016).

  19. 19.

    Khan, M. et al. Embryonic stem cell-derived exosomes promote endogenous repair mechanisms and enhance cardiac function following myocardial infarction. Circ. Res. 117, 52–64 (2015).

  20. 20.

    Bian, S. et al. Extracellular vesicles derived from human bone marrow mesenchymal stem cells promote angiogenesis in a rat myocardial infarction model. J. Mol. Med. 92, 387–397 (2014).

  21. 21.

    Mackie, A. R. et al. Sonic hedgehog-modified human CD34+ cells preserve cardiac function after acute myocardial infarction. Circ. Res. 111, 312–321 (2012).

  22. 22.

    Chong, J. J. H. et al. Human embryonic-stem-cell-derived cardiomyocytes regenerate non-human primate hearts. Nature 510, 273–277 (2014).

  23. 23.

    Laflamme, M. A. et al. Cardiomyocytes derived from human embryonic stem cells in pro-survival factors enhance function of infarcted rat hearts. Nat. Biotechnol. 25, 1015–1024 (2007).

  24. 24.

    Lee, A. S., Tang, C., Rao, M. S., Weissman, I. L. & Wu, J. C. Tumorigenicity as a clinical hurdle for pluripotent stem cell therapies. Nat. Med. 19, 998–1004 (2013).

  25. 25.

    Chong, J. J. H. et al. Human embryonic-stem-cell-derived cardiomyocytes regenerate non-human primate hearts. Nature 510, 273–277 (2014).

  26. 26.

    Serpooshan, V. & Wu, S. M. Patching up broken hearts: cardiac cell therapy gets a bioengineered boost. Cell Stem Cell 15, 671–673 (2014).

  27. 27.

    Anderson, M. E., Goldhaber, J. I., Houser, S. I., Puceat, M. & Sussman, M. A. Embryonic stem cell-derived cardiac myocytes are not ready for human trials. Circ. Res. 115, 335–338 (2014).

  28. 28.

    Waldenström, A., Gennebäck, N., Hellman, U., Ronquist, G. & Minetti, C. Cardiomyocyte microvesicles contain DNA/RNA and convey biological messages to target cells. PLoS ONE 7, e34653 (2012).

  29. 29.

    Garcia, N. A., Moncayo-Arlandi, J., Sepulveda, P. & Diez-Juan, A. Cardiomyocyte exosomes regulate glycolytic flux in endothelium by direct transfer of GLUT transporters and glycolytic enzymes. Cardiovasc. Res. 109, 397–408 (2016).

  30. 30.

    Wang, X. et al. Cardiomyocytes mediate anti-angiogenesis in type 2 diabetic rats through the exosomal transfer of miR-320 into endothelial cells. J. Mol. Cell. Cardiol. 74, 139–150 (2014).

  31. 31.

    Zhang, X. et al. Hsp20 functions as a novel cardiokine in promoting angiogenesis via activation of VEGFR2. PLoS ONE 7, e32765 (2012).

  32. 32.

    Kishore, R. & Khan, M. More than tiny sacks: stem cell exosomes as cell-free modality for cardiac repair. Circ. Res. 118, 330–343 (2016).

  33. 33.

    Chernyshev, V. S. et al. Size and shape characterization of hydrated and desiccated exosomes. Anal. Bioanal. Chem. 407, 3285–3301 (2015).

  34. 34.

    Mateescu, B. et al. Obstacles and opportunities in the functional analysis of extracellular vesicle RNA—an ISEV position paper. J. Extracell. Vesicles 6, 1286095 (2017).

  35. 35.

    Witwer, K. W. et al. Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. J. Extracell. Vesicles 2, 20360 (2013).

  36. 36.

    Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-Seq data with DESeq2. Genome Biol. 15, 550 (2014).

  37. 37.

    Yang, B. et al. The muscle-specific microRNA miR-1 regulates cardiac arrhythmogenic potential by targeting GJA1 and KCNJ2. Nat. Med. 13, 486–491 (2007).

  38. 38.

    Tian, Y. et al. A microRNA–Hippo pathway that promotes cardiomyocyte proliferation and cardiac regeneration in mice. Sci. Transl. Med. 7, 279ra38 (2015).

  39. 39.

    Carè, A. et al. MicroRNA-133 controls cardiac hypertrophy. Nat. Med. 13, 613–618 (2007).

  40. 40.

    Lewis, B. P. et al. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120, 15–20 (2005).

  41. 41.

    Garcia, D. M. et al. Weak seed-pairing stability and high target-site abundance decrease the proficiency of lsy-6 and other microRNAs. Nat. Struct. Mol. Biol. 18, 1139–1146 (2011).

  42. 42.

    The Gene Ontogoly Consortium. Gene Ontology Consortium: going forward. Nucleic Acids Res. 43, D1049–D1056 (2015).

  43. 43.

    Ashburner, M. et al. Gene ontology: tool for the unification of biology. Nat. Genet. 25, 25–29 (2000).

  44. 44.

    Wallace, D. G. & Rosenblatt, J. Collagen gel systems for sustained delivery and tissue engineering. Adv. Drug Deliv. Rev. 55, 1631–1649 (2003).

  45. 45.

    Protze, S. I. et al. Sinoatrial node cardiomyocytes derived from human pluripotent cells function as a biological pacemaker. Nat. Biotechnol. 35, 56–68 (2016).

  46. 46.

    Eng, G. et al. Autonomous beating rate adaptation in human stem cell-derived cardiomyocytes. Nat. Commun. 7, 10312 (2016).

  47. 47.

    Sontag, S. et al. Modelling IRF8 deficient human hematopoiesis and dendritic cell development with engineered iPS cells. Stem Cells 35, 898–908 (2017).

  48. 48.

    Chen, H.-S. V., Kim, C. & Mercola, M. Electrophysiological challenges of cell-based myocardial repair. Circulation 120, 2496–2508 (2009).

  49. 49.

    Zhu, X. et al. Comprehensive toxicity and immunogenicity studies reveal minimal effects in mice following sustained dosing of extracellular vesicles derived from HEK293T cells. J. Extracell. Vesicles 6, 1324730 (2017).

  50. 50.

    Hou, D. et al. Radiolabeled cell distribution after intramyocardial, intracoronary, and interstitial retrograde coronary venous delivery: implications for current clinical trials. Circulation 112, I150–I156 (2005).

  51. 51.

    Seif-Naraghi, S. B. et al. Safety and efficacy of an injectable extracellular matrix hydrogel for treating myocardial infarction. Sci. Transl. Med. 5, 173ra25 (2013).

  52. 52.

    Lang, N. et al. A blood-resistant surgical glue for minimally invasive repair of vessels and heart defects. Sci. Transl. Med. 6, 218ra6 (2014).

  53. 53.

    Si-Tayeb, K. et al. Highly efficient generation of human hepatocyte-like cells from induced pluripotent stem cells. Hepatology 51, 297–305 (2010).

  54. 54.

    Ibrahim, A. G.-E., Cheng, K. & Marbán, E. Exosomes as critical agents of cardiac regeneration triggered by cell therapy. Stem Cell Rep. 2, 606–619 (2014).

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We thank M. Moore (Memorial Sloan Kettering Cancer Center) for making the particle-tracking instrument (NanoSight) available, and S. R. Ambati and A. Saxena (Memorial Sloan Kettering Cancer Center) for technical help. We thank Q. Li for performing animal surgeries, R. Liu and L. Zaurov for assistance with animal echocardiograms, and S. Halligan for coordinating the animal work. We thank D. Teles, N. Kim and A. Pluchinksky for assistance with the experiments. We thank B. Fine for valuable discussions on the manuscript. We gratefully acknowledge funding for this work by the NIH (HL076485, EB002520, EB17103 and GM007367), NYSTEM (C028119), the NIA (F30 AG047748) and the Lisa and Mark Schwartz Program for Reversing Heart Failure.

Author information

Author notes

  1. These authors contributed equally: Bohao Liu, Benjamin W. Lee.


  1. College of Physicians and Surgeons, Columbia University, New York, NY, USA

    • Bohao Liu
    • , Benjamin W. Lee
    • , Rebecca Williamson
    •  & Jordan Metz
  2. Department of Medicine, Columbia University, New York, NY, USA

    • Bohao Liu
    • , Koki Nakanishi
    • , Mariko Kanai
    • , Shunichi Homma
    • , Veli K. Topkara
    •  & Gordana Vunjak-Novakovic
  3. Department of Biomedical Engineering, Columbia University, New York, NY, USA

    • Benjamin W. Lee
    • , Aranzazu Villasante
    • , Jinho Kim
    • , Lynn Bi
    •  & Gordana Vunjak-Novakovic
  4. Department of Pathology and Cell Biology, Columbia University, New York, NY, USA

    • Rebecca Williamson
    • , Kristy Brown
    •  & Gilbert Di Paolo
  5. Department of Systems Biology, Columbia University, New York, NY, USA

    • Jordan Metz
    •  & Peter A. Sims
  6. Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA

    • Peter A. Sims


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B.L., B.W.L., G.D.P., S.H., P.A.S., V.K.T. and G.V.-N. designed the study. B.L., B.W.L., K.N., A.V., R.W., J.K., M.K., L.B. and K.B. performed the experiments. B.L., B.W.L. and J.M. analysed the data. B.L., B.W.L., P.A.S., V.K.T. and G.V.-N. wrote the manuscript.

Competing interests

The authors declare no competing interests.

Corresponding author

Correspondence to Gordana Vunjak-Novakovic.

Supplementary information

  1. Supplementary Information

    Supplementary figures and tables

  2. Reporting Summary

  3. Supplementary Dataset

    Exosome miRNA sequencing, miRNA search targets, gene ontology, echocardiography data and correlation analysis.