Oral Presentation Australia and New Zealand Society for Extracellular Vesicles Conference 2023

Stem cell-derived nanovesicles (scNVs) as a scalable, cell reprograming therapeutic strategy for cardiac repair (#31)

Jonathan Lozano 1 2 3 , Jarmon Lees 4 5 , Alin Rai 1 2 3 6 , Jonathon Cross 2 , Haoyun Fang 1 7 , Shiang Y Lim 4 5 , David W Greening 1 2 3 6 7
  1. Department of Cardiovascular Research Translation and Implementation. , Baker Heart and Diabetes Institute, Melbourne, Victoria, Australia
  2. Department of Molecular Proteomics, Baker Heart and Diabetes Institute, Melbourne, Victoria, Australia
  3. Department of Microbiology, Anatomy, Physiology and Pharmacology, School of Agriculture, Biomedicine and Environment, , La Trobe University, Bundoora, Victoria, Australia
  4. Cardiac Regeneration Laboratory , O'Brien Institute Department, St Vincent's Institute of Medical Research., Melbourne, Victoria, Australia
  5. Department of Surgery, University of Melbourne, St Vincent Hospital, Melbourne, Victoria, Australia
  6. Baker Department of Cardiometabolic Health, University of Melbourne, Melbourne, Victoria, Australia
  7. Central Clinical School, Monash University, Melbourne, Victoria, Australia

Stem cell-derived extracellular vesicles (EVs) are a potential strategy for cardiac tissue repair following ischaemia-reperfusion injury. However, scalable generation limits their clinical application and remains an unmet clinical need. Here, we propose stem cell-derived nanovesicles (NVs) as a surrogate to natural EVs towards reproducible, rapid, and scalable deliverable therapy for cardiac repair.

 

NVs were generated from different human-induced pluripotent stem cells through serial size-based membrane extrusion in large quantities (yield 900× natural EVs). NVs were isolated using density-gradient separation (NVsd=1.13 g/mL), are spherical in shape (~100 nm), morphologically intact, and readily taken up by human cardiomyocytes, primary cardiac fibroblasts, and endothelial cells. Mass spectrometry-based proteomics revealed that NVs captured the dynamic proteome of parental cells, including pluripotency markers (LIN28A, OCT4), and regulators of cardiac repair processes. Functionally, single-dose NVs significantly promoted tubule formation of endothelial cells (angiogenesis) (p<0.05) and survival of cardiomyocytes exposed to hypoxia (p<0.0001), as well as attenuate activation of cardiac fibroblasts (p<0.0001). In human cardiac organoids we demonstrate NVs preserve overall contractility function; total contraction duration, time to peak, relaxation time and ratio of relaxation to contraction velocities (p<0.05). Quantitative proteome profiling of cells and organoids proteomes following NV treatment revealed upregulation of pro-survival network (MDH2, LRPPRC, NIPSNAP1), tissue repair (HSP70, CYFIP1), pro-angiogenic (FARSA, ECE1, RRAS), and cardiac function (XIRP1, SLMAP, MYH6, CTNNA1, NDUFS2, GPD2).

 

In summary, this study showcases a scalable approach to generating functional nanovesicles, highlights their multimodal therapeutic potential, and identifies key players involved in cardiac repair processes.