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.