Extracellular vesicles derived from human bone marrow mesenchymal stem cells promote angiogenesis in a rat myocardial infarction model

Mesenchymal stem cells (MSCs) have been increasingly tested experimentally and clinically for cardiac repair. However, the underlying mechanisms remain controversial due to the poor viability and considerable death of the engrafted cells in the infracted myocardium. Recent reports have suggested that extracellular vesicles (EVs) released by MSCs have angiogenesis-promoting activity; however, the therapeutic effect of MSC-EVs on an ischemic heart is unclear. In the present study, we reported that MSCs could release a large quantity of EVs around 100 nm in diameter upon hypoxia stimulation though the majority of the cells had not experienced apoptosis. MSC-EVs could be promptly uptaken by human umbilical vein endothelial cells, and the internalization resulted in dose-dependent enhancement of in vitro proliferation, migration, and tube formation of endothelial cells. Using an acute myocardial infarction rat model, we found that intramyocardial injection of MSC-EVs markedly enhanced blood flow recovery, in accordance with reduced infarct size and preserved cardiac systolic and diastolic performance compared to those treated with PBS. These data suggest that like MSCs, MSC-EVs could also protect cardiac tissue from ischemic injury at least by means of promoting blood vessel formation, though further detailed investigations should be performed to define the functionality of MSC-EVs.Key messagesMSCs released extracellular vesicles (EVs) upon hypoxia stimulation.MSC-EVs were a mixture of microvesicles and exosomes.MSC-EVs could be promptly uptaken by human umbilical vein endothelial cells.MSC-EVs promoted neoangiogenesis in vitro and in vivo.MSC-EVs preserved cardiac performance in an AMI model.

[1]  D. Pisetsky,et al.  The induction of matrix metalloproteinase and cytokine expression in synovial fibroblasts stimulated with immune cell microparticles. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Jin Yuan,et al.  Mechanism of TNF-α autocrine effects in hypoxic cardiomyocytes: initiated by hypoxia inducible factor 1α, presented by exosomes. , 2012, Journal of molecular and cellular cardiology.

[3]  Oscar K Lee,et al.  Hypoxia promotes proliferation and osteogenic differentiation potentials of human mesenchymal stem cells , 2012, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[4]  C. Xinaris,et al.  Thyroid hormone attenuates cardiac remodeling and improves hemodynamics early after acute myocardial infarction in rats. , 2007, European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery.

[5]  Koichi Hattori,et al.  Adipocyte-derived microvesicles are associated with multiple angiogenic factors and induce angiogenesis in vivo and in vitro. , 2010, Endocrinology.

[6]  Ling Wei,et al.  Transplantation of hypoxia-preconditioned mesenchymal stem cells improves infarcted heart function via enhanced survival of implanted cells and angiogenesis. , 2008, The Journal of thoracic and cardiovascular surgery.

[7]  Paolo Ubezio,et al.  Heterogeneous cell response to topotecan in a CFSE‐based proliferation test , 2004, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[8]  A. Keating,et al.  Bone Marrow-derived Mesenchymal Stromal Cells Express Cardiac-specific Markers, Retain the Stromal Phenotype, and Do Not Become Functional Cardiomyocytes in Vitro Departments of a Physiology And , 2022 .

[9]  Luca Sterpone,et al.  Microvesicles Derived from Adult Human Bone Marrow and Tissue Specific Mesenchymal Stem Cells Shuttle Selected Pattern of miRNAs , 2010, PloS one.

[10]  G. Camussi,et al.  Paracrine/endocrine mechanism of stem cells on kidney repair: role of microvesicle-mediated transfer of genetic information , 2010, Current opinion in nephrology and hypertension.

[11]  G. Grau,et al.  In vitro generation of endothelial microparticles and possible prothrombotic activity in patients with lupus anticoagulant. , 1999, The Journal of clinical investigation.

[12]  H. Duan,et al.  Adenoviral gene transfer of sphingosine kinase 1 protects heart against ischemia/reperfusion-induced injury and attenuates its postischemic failure. , 2007, Human gene therapy.

[13]  M. Kesimer,et al.  Morphologic and proteomic characterization of exosomes released by cultured extravillous trophoblast cells. , 2011, Experimental cell research.

[14]  M. Bennett,et al.  Signalling from dead cells drives inflammation and vessel remodelling. , 2012, Vascular pharmacology.

[15]  Aled Clayton,et al.  Isolation and Characterization of Exosomes from Cell Culture Supernatants and Biological Fluids , 2006, Current protocols in cell biology.

[16]  Usha Nekanti,et al.  Increased Proliferation and Analysis of Differential Gene Expression in Human Wharton's Jelly-derived Mesenchymal Stromal Cells under Hypoxia , 2010, International journal of biological sciences.

[17]  M. Johansson,et al.  Hypoxia triggers a proangiogenic pathway involving cancer cell microvesicles and PAR-2–mediated heparin-binding EGF signaling in endothelial cells , 2011, Proceedings of the National Academy of Sciences.

[18]  D. Hwang,et al.  Proteomic analysis of microvesicles derived from human mesenchymal stem cells. , 2012, Journal of proteome research.

[19]  E. Lo,et al.  A new penumbra: transitioning from injury into repair after stroke , 2008, Nature Medicine.

[20]  Robert A. Kloner,et al.  Systemic Delivery of Bone Marrow–Derived Mesenchymal Stem Cells to the Infarcted Myocardium: Feasibility, Cell Migration, and Body Distribution , 2003, Circulation.

[21]  M. Michael,et al.  Hypoxic enhancement of exosome release by breast cancer cells , 2012, BMC Cancer.

[22]  C. Hamm,et al.  Stem cells in myocardial infarction: from bench to bedside , 2009, Heart.

[23]  J. Ryan,et al.  Exosomal Signaling during Hypoxia Mediates Microvascular Endothelial Cell Migration and Vasculogenesis , 2013, PloS one.

[24]  X. Chen,et al.  Microvesicle-mediated Transfer of MicroRNA-150 from Monocytes to Endothelial Cells Promotes Angiogenesis* , 2013, The Journal of Biological Chemistry.

[25]  E. Yeh,et al.  Molecular Imaging of Mesenchymal Stem Cell: Mechanistic Insight Into Cardiac Repair After Experimental Myocardial Infarction , 2012, Circulation. Cardiovascular imaging.

[26]  Alessandro Busca,et al.  Mesenchymal stem cell-derived microvesicles protect against acute tubular injury. , 2009, Journal of the American Society of Nephrology : JASN.

[27]  Yang Yang,et al.  Microvesicles derived from human umbilical cord mesenchymal stem cells stimulated by hypoxia promote angiogenesis both in vitro and in vivo. , 2012, Stem cells and development.

[28]  J. Freyssinet,et al.  Membrane microparticles: two sides of the coin. , 2005, Physiology.

[29]  J Ratajczak,et al.  Embryonic stem cell-derived microvesicles reprogram hematopoietic progenitors: evidence for horizontal transfer of mRNA and protein delivery , 2006, Leukemia.

[30]  G. Camussi,et al.  Role of mesenchymal stem cell-derived microvesicles in tissue repair , 2013, Pediatric Nephrology.

[31]  R. Andriantsitohaina,et al.  Microparticles Carrying Sonic Hedgehog Favor Neovascularization through the Activation of Nitric Oxide Pathway in Mice , 2010, PloS one.

[32]  M. Goumans,et al.  Human mesenchymal stem cell-conditioned medium improves cardiac function following myocardial infarction. , 2011, Stem cell research.

[33]  P. Quesenberry,et al.  Stem cell plasticity revisited: the continuum marrow model and phenotypic changes mediated by microvesicles. , 2010, Experimental hematology.

[34]  G. Camussi,et al.  Microvesicles derived from human adult mesenchymal stem cells protect against ischaemia-reperfusion-induced acute and chronic kidney injury. , 2011, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[35]  J. Mesirov,et al.  Carcinoma-associated fibroblast-like differentiation of human mesenchymal stem cells. , 2008, Cancer research.

[36]  Tian Sheng Chen,et al.  Mesenchymal stem cell secretes microparticles enriched in pre-microRNAs , 2009, Nucleic acids research.

[37]  A. Hagège,et al.  Comparison of the effects of fetal cardiomyocyte and skeletal myoblast transplantation on postinfarction left ventricular function. , 2000, The Journal of thoracic and cardiovascular surgery.

[38]  Gerard Pasterkamp,et al.  Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury. , 2010, Stem cell research.

[39]  M. Taupitz,et al.  In vivo visualization of locally transplanted mesenchymal stem cells in the severely injured muscle in rats. , 2008, Tissue engineering. Part A.

[40]  J. Ingwall,et al.  Evidence supporting paracrine hypothesis for Akt‐modified mesenchymal stem cell‐mediated cardiac protection and functional improvement , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[41]  X. Qin,et al.  Paracrine action enhances the effects of autologous mesenchymal stem cell transplantation on vascular regeneration in rat model of myocardial infarction. , 2005, The Annals of thoracic surgery.

[42]  Hamid Cheshmi.  Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers , 2011 .

[43]  Arjun Deb,et al.  Mesenchymal stem cells overexpressing Akt dramatically repair infarcted myocardium and improve cardiac function despite infrequent cellular fusion or differentiation. , 2006, Molecular therapy : the journal of the American Society of Gene Therapy.

[44]  A. Brill,et al.  Platelet-derived microparticles induce angiogenesis and stimulate post-ischemic revascularization. , 2004, Cardiovascular research.

[45]  A. Lavrentieva,et al.  Effects of hypoxic culture conditions on umbilical cord-derived human mesenchymal stem cells , 2010, Cell Communication and Signaling.

[46]  J. Fehrenbacher,et al.  Signal transducer and activator of transcription 3-stimulated hypoxia inducible factor-1alpha mediates estrogen receptor-alpha-induced mesenchymal stem cell vascular endothelial growth factor production. , 2009, The Journal of thoracic and cardiovascular surgery.

[47]  S. Korsmeyer,et al.  Cell Death Critical Control Points , 2004, Cell.

[48]  Luigi Biancone,et al.  Exosomes/microvesicles as a mechanism of cell-to-cell communication. , 2010, Kidney international.

[49]  Luigi Biancone,et al.  Endothelial progenitor cell derived microvesicles activate an angiogenic program in endothelial cells by a horizontal transfer of mRNA. , 2007, Blood.

[50]  J. Rouleau,et al.  Long-Term Effects of Carvedilol on Left Ventricular Function, Remodeling, and Expression of Cardiac Cytokines After Large Myocardial Infarction in the Rat , 2002, Journal of cardiovascular pharmacology.