Enhancement of neovascularization with cord blood CD133⁺ cell-derived endothelial progenitor cell transplantation

Summary

The endothelial progenitor cells (EPCs) are responsible for postnatal vasculogenesis in physiological and pathological neovascularization and have been used for attenuating ischemic diseases. However, EPCs from umbilical cord blood (CB) were not well understood and the homing mechanisms of EPCs remain unclear. To determine the potential application of CB-derived EPCs, we established a culture system to induce the differentiation of CB cells into EPCs. Purified CB CD133⁺ cells proliferated and, after further vascular endothelial growth factor receptor 2 (VEGFR-2) antibody purification, differentiated into EPCs expressing endothelial markers, such as VE-cadherin, VEGFR-2, CD31, von Willebrand factor (vWF) and WeibelPalade bodies. These cells could also take up acetylated lower density lipoprotein (Ac-LDL) and bind Ulex europaeus agglutinin-1 (UEA-1).When expanded EPCs were transplanted via tail vein into nude mice, they incorporated into capillary networks in ischemic hindlimb, augmented neovascularization, and improved ischemic limb salvage. In addition, in ischemic tissue, there were elevated expressions of VEGF and stromal derived factor 1α???????????(SDF-1α), both of which had chemotactic effect on EPCs. Moreover, P-/E-selectins was found on mouse ischemic endothelium and P-selectin glycoprotein ligand-1 (PSGL-1) on CB-derived EPCs. Neutralizing antibody against PSGL-1 blocked the homing of EPCs to ischemic area by 61%. These results demonstrate that CB CD133+ cell-derived EPCs can be applied for therapeutic neovascularization in ischemic diseases, and reveal important roles of chemoattractants and adhesive molecules in the homing of EPCs.

Keywords

Endothelial progenitor cells - cord blood - CD133 - transplantation - homing

• References

• 1 Buschmann I, Schaper W. The pathophysiology of the collateral circulation (arteriogenesis). J Pathol 2000; 190: 338-42.
• 2 Risau W. Mechanisms of angiogenesis. Nature 1997; 386: 671-4.
• 3 Carmeliet P. Mechanisms of angiogenesis and arteriogenesis. Nat Med 2000; 06: 389-95.
• 4 Asahara T, Murohara T, Sullivan A. et al. Isolation of putative progenitor endothelial cells for angiogenesis. Science 1997; 275: 964-7.
• 5 Murayama T, Tepper OM, Silver M. et al. Determination of bone marrow-derived endothelial progenitor cell significance in angiogenic growth factor-induced neovascularization in vivo . Exp Hematol 2002; 30: 967-72.
• 6 Asahara T, Masuda H, Takahashi T. et al. Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circ Res 1999; 85: 221-8.
• 7 Lyden D, Hattori K, Dias S. et al. Impaired recruitment of bone-marrow-derived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth. Nat Med 2001; 07: 1194-201.
• 8 Zhang ZG, Zhang L, Jiang Q. et al. Bone marrow-derived endothelial progenitor cells participate in cerebral neovascularization after focal cerebral ischemia in the adult mouse. Circ Res 2002; 90: 284-8.
• 9 Hill JM, Zalos G, Halcox JP. et al. Circulating endothelial progenitor cells, vascular function, and cardiovascular risk. N Engl J Med 2003; 348: 593-600.
• 10 Couffinhal T, Silver M, Kearney M. et al. Impaired collateral vessel development associated with reduced expression of vascular endothelial growth factor in ApoE−/−mice. Circulation 1999; 99: 3188-98.
• 11 Rivard A, Fabre JE, Silver M. et al. Agedependent impairment of angiogenesis. Circulation 1999; 99: 111-20.
• 12 Tepper OM, Galiano RD, Capla JM. et al. Human endothelial progenitor cells from type II diabetics exhibit impaired proliferation, adhesion, and incorporation into vascular structures. Circulation 2002; 106: 2781-6.
• 13 Freedman SB, Isner JM. Therapeutic angiogenesis for ischemic cardiovascular disease. J Mol Cell Cardiol 2001; 33: 379-93.
• 14 Hamawy AH, Lee LY, Crystal RG. et al. Cardiac angiogenesis and gene therapy: a strategy for myocardial revascularization. Curr Opin Cardiol 1999; 14: 515-22.
• 15 Henry TD, Abraham JA. Review of preclinical and clinical results with vascular endothelial growth factors for therapeutic angiogenesis. Curr Interv Cardiol Rep 2000; 02: 228-41.
• 16 Ito WD, Arras M, Winkler B. et al. Monocyte chemotactic protein-1 increases collateral and peripheral conductance after femoral artery occlusion. Circ Res 1997; 80: 829-37.
• 17 Asahara T, Takahashi T, Masuda H. et al. VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells. EMBO J 1999; 18: 3964-72.
• 18 Gill M, Dias S, Hattori K. et al. Vascular trauma induces rapid but transient mobilization of VEGFR2(+)AC133(+) endothelial precursor cells. Circ Res 2001; 88: 167-74.
• 19 Kalka C, Tehrani H, Laudenberg B. et al. VEGF gene transfer mobilizes endothelial progenitor cells in patients with inoperable coronary disease. Ann Thorac Surg 2000; 70: 829-34.
• 20 Murohara T, Ikeda H, Duan J. et al. Transplanted cord blood-derived endothelial precursor cells augment postnatal neovascularization. J Clin Invest 2000; 105: 1527-36.
• 21 Kalka C, Masuda H, Takahashi T. et al. Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci U S A 2000; 97: 3422-7.
• 22 Kawamoto A, Gwon HC, Iwaguro H. et al. Therapeutic potential of ex vivo expanded endothelial progenitor cells for myocardial ischemia. Circulation 2001; 103: 634-7.
• 23 Schatteman GC, Hanlon HD, Jiao C. et al. Blood-derived angioblasts accelerate bloodflow restoration in diabetic mice. J Clin Invest 2000; 106: 571-8.
• 24 Kocher AA, Schuster MD, Szabolcs MJ. et al. Neovascularization of ischemic myocardium by human bone-marrow-derived angioblasts prevents cardiomyocyte apoptosis, reduces remodeling and improves cardiac function. Nat Med 2001; 07: 430-6.
• 25 Shintani S, Murohara T, Ikeda H. et al. Augmentation of postnatal neovascularization with autologous bone marrow transplantation. Circulation 2001; 103: 897-903.
• 26 Kaushal S, Amiel GE, Guleserian KJ. et al. Functional small-diameter neovessels created using endothelial progenitor cells expanded ex vivo. Nat Med 2001; 07: 1035-40.
• 27 Shi Q, Rafii S, Wu MH. et al. Evidence for circulating bone marrow-derived endothelial cells. Blood 1998; 92: 362-7.
• 28 Takahashi T, Kalka C, Masuda H. et al. Ischemia-and cytokine-induced mobilization of bone marrow-derived endothelial progenitor cells for neovascularization. Nat Med 1999; 05: 434-8.
• 29 Quirici N, Soligo D, Caneva L. et al. Differentiation and expansion of endothelial cells from human bone marrow CD133(+) cells. Br J Haematol 2001; 115: 186-94.
• 30 Peichev M, Naiyer AJ, Pereira D. et al. Expression of VEGFR-2 and AC133 by circulating human CD34(+) cells identifies a population of functional endothelial precursors. Blood 2000; 95: 952-8.
• 31 Harraz M, Jiao C, Hanlon HD. et al. CD34blood-derived human endothelial cell progenitors. Stem Cells 2001; 19: 304-12.
• 32 Gehling UM, Ergun S, Schumacher U. et al. In vitro differentiation of endothelial cells from AC133-positive progenitor cells. Blood 2000; 95: 3106-12.
• 33 Rehman J, Li J, Orschell CM. et al. Peripheral blood “endothelial progenitor cells” are derived from monocyte/macrophages and secrete angiogenic growth factors. Circulation 2003; 107: 1164-9.
• 34 Pesce M, Orlandi A, Iachininoto MG. et al. Myoendothelial differentiation of human umbilical cord blood-derived stem cells in ischemic limb tissues. Circ Res 2003; 93: e51-62.
• 35 Eggermann J, Kliche S, Jarmy G. et al. Endothelial progenitor cell culture and differentiation in vitro: a methodological comparison using human umbilical cord blood. Cardiovasc Res 2003; 58: 478-86.
• 36 Hatzopoulos AK, Folkman J, Vasile E. et al. Isolation and characterization of endothelial progenitor cells from mouse embryos. Development 1998; 125: 1457-68.
• 37 Vajkoczy P, Blum S, Lamparter M. et al. Multistep nature of microvascular recruitment of ex vivo-expanded embryonic endothelial progenitor cells during tumor angiogenesis. J Exp Med 2003; 197: 1755-65.
• 38 Miraglia S, Godfrey W, Yin AH. et al. A novel five-transmembrane hematopoietic stem cell antigen: isolation, characterization, and molecular cloning. Blood 1997; 90: 5013-21.
• 39 Yin AH, Miraglia S, Zanjani ED. et al. AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 1997; 90: 5002-12.
• 40 Vaziri H, Dragowska W, Allsopp RC. et al. Evidence for a mitotic clock in human hematopoietic stem cells: loss of telomeric DNA with age. Proc Natl Acad Sci U S A 1994; 91: 9857-60.
• 41 Couffinhal T, Silver M, Zheng LP. et al. Mouse model of angiogenesis. Am J Pathol 1998; 152: 1667-79.
• 42 Weissman IL. Stem cells: units of development, units of regeneration, and units in evolution. Cell 2000; 100: 157-68.
• 43 Brizzi MF, Garbarino G, Rossi PR. et al. Interleukin 3 stimulates proliferation and triggers endothelial-leukocyte adhesion molecule 1 gene activation of human endothelial cells. J Clin Invest 1993; 91: 2887-92.
• 44 Mazo IB, Gutierrez-Ramos JC, Frenette PS. et al. Hematopoietic progenitor cell rolling in bone marrow microvessels: parallel contributions by endothelial selectins and vascular cell adhesion molecule 1. J Exp Med 1998; 188: 465-74.
• 45 Yamaguchi J, Kusano KF, Masuo O. et al. Stromal cell-derived factor-1 effects on ex vivo expanded endothelial progenitor cell recruitment for ischemic neovascularization. Circulation 2003; 107: 1322-8.
• 46 Semenza GL. Hypoxia-inducible factor 1: master regulator of O2 homeostasis. Curr Opin Genet Dev 1998; 08: 588-94.
• 47 Nagaya N, Kangawa K, Kanda M. et al. Hybrid Cell–Gene Therapy for Pulmonary Hypertension Based on Phagocytosing Action of Endothelial Progenitor Cells. Circulation 2003; 108: 889-95.