title: Up-regulation of miR-210 by vascular endothelial growth factor in ex vivo expanded CD34+ cells enhances cell-mediated angiogenesis.
abstract: Ex vivo culture has been proposed as a means to augment and repair autologous cells in patients with chronic diseases, but the mechanisms governing improvement in cell function are not well understood. Although microRNAs (miRs) are increasingly appreciated as key regulators of cellular function, a role for these factors in CD34+ cell-mediated angiogenesis has not been elucidated. Vascular endothelial growth factor (VEGF) was previously shown to induce expression of certain miRs associated with angiogenesis in endothelial cells and promote survival and number of vascular colony forming units of haematopoietic stem cells (HSCs). We sought to evaluate the role of VEGF in expansion and angiogenic function of CD34+ cells and to identify specific miRs associated with angiogenic properties of expanded cells. Umbilical cord blood CD34+ cells were effectively expanded (18- to 22-fold) in culture medium containing stem cell factor (SCF), Flt-3 ligand (Flt-3), thrombopoietin (TPO) and interleukin-6 (IL-6) with (postEX/+VEGF) and without VEGF (postEX/noVEGF). Tube formation in matrigel assay and tissue perfusion/capillary density in mice ischaemic hindlimb were significantly improved by postEX/+VEGF cells compared with fresh CD34+ and postEX/noVEGF cells. MiR-210 expression was significantly up-regulated in postEX/+VEGF cells. MiR-210 inhibitor abrogated and 210 mimic recapitulated the pro-angiogenic effects by treatment of postEX/+VEGF and postEX/noVEGF cells respectively. Collectively, these observations highlight a critical role for VEGF in enhancing the angiogenic property of expanded cells, and identify miR-210 as a potential therapeutic target to enhance CD34+ stem cell function for the treatment of ischaemic vascular disease.
authors.affiliation: Department of Medicine, Case Cardiovascular Research Institute, University Hospital Case Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH 44106-5038, USA.
authors.forename: Mohamad Amer
authors.initials: MA
authors.lastname: Alaiti
authors.forename: Masakazu
authors.initials: M
authors.lastname: Ishikawa
authors.forename: Haruchika
authors.initials: H
authors.lastname: Masuda
authors.forename: Daniel I
authors.initials: DI
authors.lastname: Simon
authors.forename: Mukesh K
authors.initials: MK
authors.lastname: Jain
authors.forename: Takayuki
authors.initials: T
authors.lastname: Asahara
authors.forename: Marco A
authors.initials: MA
authors.lastname: Costa
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000378:metabolism*; D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology; D005786:Gene Expression Regulation; D020868:Gene Silencing; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D018919:Neovascularization, Physiologic; D019089:Stem Cell Factor; D013926:Thrombopoietin / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
chemical_list: D043925:Angiogenesis Inducing Agents; D018952:Antigens, CD34; D015850:Interleukin-6; C543800:MIRN210 microRNA, human; D008565:Membrane Proteins; D035683:MicroRNAs; D019089:Stem Cell Factor; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C084881:flt3 ligand protein; D013926:Thrombopoietin