@article{83c01ee2a91742d29d96b592bff81c04,
title = "Extracellular vesicles from differentiated stem cells contain novel proangiogenic miRNAs and induce angiogenic responses at low doses",
abstract = "Extracellular vesicles (EVs) released from healthy endothelial cells (ECs) have shown potential for promoting angiogenesis, but their therapeutic efficacy remains poorly understood. We have previously shown that transplantation of a human embryonic stem cell–derived endothelial cell product (hESC-ECP), promotes new vessel formation in acute ischemic disease in mice, likely via paracrine mechanism(s). Here, we demonstrated that EVs from hESC-ECPs (hESC-eEVs) significantly increased EC tube formation and wound closure in vitro at ultralow doses, whereas higher doses were ineffective. More important, EVs isolated from the mesodermal stage of the differentiation (hESC-mEVs) had no effect. Small RNA sequencing revealed that hESC-eEVs have a unique transcriptomic profile and are enriched in known proangiogenic microRNAs (miRNAs, miRs). Moreover, an in silico analysis identified three novel hESC-eEV-miRNAs with potential proangiogenic function. Differential expression analysis suggested that two of those, miR-4496 and miR-4691-5p, are highly enriched in hESC-eEVs. Overexpression of miR-4496 or miR-4691-5p resulted in increased EC tube formation and wound closure in vitro, validating the novel proangiogenic function of these miRNAs. In summary, we demonstrated that hESC-eEVs are potent inducers of EC angiogenic response at ultralow doses and contain a unique EV-associated miRNA repertoire, including miR-4496 and miR-4691-5p, with novel proangiogenic function.",
keywords = "angiogenesis, delivery, dose response, endothelial cell differentiation, extracellular vesicles, microRNA, miR-4496, miR-4691-5p, myocardial infarction, vasculature",
author = "Despoina Kesidou and Matthew Bennett and Monteiro, {Jo{\~a}o P.} and McCracken, {Ian R.} and Eftychia Klimi and Julie Rodor and Alison Condie and Scott Cowan and Andrea Caporali and Wit, {Jan B.M.} and Mountford, {Joanne C.} and Mairi Brittan and Abdelaziz Beqqali and Baker, {Andrew H.}",
note = "Funding Information: We acknowledge Stephen Mitchell at the School of Biological Sciences{\textquoteright} electron microscopy unit for assistance with EM. We acknowledge the support of the Wellcome MultiUser Equipment Grant ( WT104915MA ). Flow cytometry data were generated with support from the QMRI Flow Cytometry and cell sorting facility , University of Edinburgh . NTA data were generated with support from the QMRI Nanosight facility , University of Edinburgh . Sequencing data were produced with assistance from BGI. This work has made use of the resources provided by the Edinburgh Compute and Data Facility ( http://www.ecdf.ed.ac.uk/ ). Funding Information: D.K. is funded by a doctoral studentship from Medical Research Scotland (grant no. PhD-1293-2018). A.B. is supported by European Union Horizon 2020 CardioReGenix (grant no. 825670). A.H.B. is supported by the British Heart Foundation (grant no. CH/11/2/28733) and the Centre for Regenerative Medicine (grant no. CRMR/21/290009). M.B. is supported by the British Heart Foundation (grant no. FS/16/4/31831). J.R. is supported by the British Heart Foundation (grant no. RG/20/5/34796). We acknowledge Stephen Mitchell at the School of Biological Sciences{\textquoteright} electron microscopy unit for assistance with EM. We acknowledge the support of the Wellcome MultiUser Equipment Grant (WT104915MA). Flow cytometry data were generated with support from the QMRI Flow Cytometry and cell sorting facility, University of Edinburgh. NTA data were generated with support from the QMRI Nanosight facility, University of Edinburgh. Sequencing data were produced with assistance from BGI. This work has made use of the resources provided by the Edinburgh Compute and Data Facility (http://www.ecdf.ed.ac.uk/). D.K. led the experimental design, execution, and statistical analysis, with feedback and input on the interpretation of the results provided by Andrea Caporali, M.B. A.B. and A.H.B. Bioinformatics analysis was performed by D.K. and M.B. with feedback and input on the interpretation of the results provided by J.R. M.B. A.B. and A.H.B. J.P.M. provided critical technical expertise and supervision on the miRNA mimic work. I.R.M. Alison Condie, and S.C. performed and supervised embryonic stem cell differentiations required for subsequent hESC-eEV isolation. D.K. wrote the manuscript. E.K. contributed valuable insights and feedback throughout the entire project. All of the authors reviewed and edited the manuscript. All of the authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication. The authors declare the following financial interests/personal relationships which may be considered potential competing interests: D.K. J.C.M. M.B. A.B. and A.H.B. are named inventors on two patent application related to this work. The first patent was filed by the University Court of the University of Edinburgh (no. EP22386089.1) “Extracellular Vesicles That Promote Angiogenesis or Neovascularisation.” The second patent was filed by Mirabilis Therapeutics BV (no. EP22386090.9) “ANGIOGENIC MIRNAS.” Funding Information: D.K. is funded by a doctoral studentship from Medical Research Scotland (grant no. PhD-1293-2018 ). A.B. is supported by European Union Horizon 2020 CardioReGenix (grant no. 825670 ). A.H.B. is supported by the British Heart Foundation (grant no. CH/11/2/28733 ) and the Centre for Regenerative Medicine (grant no. CRMR/21/290009 ). M.B. is supported by the British Heart Foundation (grant no. FS/16/4/31831 ). J.R. is supported by the British Heart Foundation (grant no. RG/20/5/34796 ). Publisher Copyright: {\textcopyright} 2023 The Authors",
year = "2024",
month = jan,
day = "3",
doi = "10.1016/j.ymthe.2023.11.023",
language = "English",
volume = "32",
pages = "185--203",
journal = "Molecular Therapy",
issn = "1525-0016",
publisher = "Cell Press",
number = "1",
}