Neovascularization Potential of Blood Outgrowth Endothelial Cells From Patients With Stable Ischemic Heart Failure Is Preserved

Background Blood outgrowth endothelial cells (BOECs) mediate therapeutic neovascularization in experimental models, but outgrowth characteristics and functionality of BOECs from patients with ischemic cardiomyopathy (ICMP) are unknown. We compared outgrowth efficiency and in vitro and in vivo functionality of BOECs derived from ICMP with BOECs from age‐matched (ACON) and healthy young (CON) controls. Methods and Results We isolated 3.6±0.6 BOEC colonies/100×106 mononuclear cells (MNCs) from 60‐mL blood samples of ICMP patients (n=45; age: 66±1 years; LVEF: 31±2%) versus 3.5±0.9 colonies/100×106 MNCs in ACON (n=32; age: 60±1 years) and 2.6±0.4 colonies/100×106 MNCs in CON (n=55; age: 34±1 years), P=0.29. Endothelial lineage (VEGFR2+/CD31+/CD146+) and progenitor (CD34+/CD133−) marker expression was comparable in ICMP and CON. Growth kinetics were similar between groups (P=0.38) and not affected by left ventricular systolic dysfunction, maladaptive remodeling, or presence of cardiovascular risk factors in ICMP patients. In vitro neovascularization potential, assessed by network remodeling on Matrigel and three‐dimensional spheroid sprouting, did not differ in ICMP from (A)CON. Secretome analysis showed a marked proangiogenic profile, with highest release of angiopoietin‐2 (1.4±0.3×105 pg/106 ICMP‐BOECs) and placental growth factor (5.8±1.5×103 pg/106 ICMP BOECs), independent of age or ischemic disease. Senescence‐associated β‐galactosidase staining showed comparable senescence in BOECs from ICMP (5.8±2.1%; n=17), ACON (3.9±1.1%; n=7), and CON (9.0±2.8%; n=13), P=0.19. High‐resolution microcomputed tomography analysis in the ischemic hindlimb of nude mice confirmed increased arteriogenesis in the thigh region after intramuscular injections of BOECs from ICMP (P=0.025; n=8) and CON (P=0.048; n=5) over vehicle control (n=8), both to a similar extent (P=0.831). Conclusions BOECs can be successfully culture‐expanded from patients with ICMP. In contrast to impaired functionality of ICMP‐derived bone marrow MNCs, BOECs retain a robust proangiogenic profile, both in vitro and in vivo, with therapeutic potential for targeting ischemic disease.

[1]  L. Lerman,et al.  Preserved Function of Late-Outgrowth Endothelial Cells in Medically Treated Hypertensive Patients Under Well-Controlled Conditions , 2014, Hypertension.

[2]  E. Creemers,et al.  Individual with Subclinical Atherosclerosis Have Impaired Proliferation of Blood Outgrowth Endothelial Cells, Which Can Be Restored by Statin Therapy , 2014, PloS one.

[3]  G. Truskey,et al.  Late-outgrowth endothelial progenitors from patients with coronary artery disease: endothelialization of confluent stromal cell layers. , 2014, Acta biomaterialia.

[4]  T. Henry,et al.  Treatment of refractory angina in patients not suitable for revascularization , 2014, Nature Reviews Cardiology.

[5]  P. Barnes,et al.  Dysfunction of Endothelial Progenitor Cells from Smokers and Chronic Obstructive Pulmonary Disease Patients Due to Increased DNA Damage and Senescence , 2013, Stem cells.

[6]  M. Berger,et al.  Functional late outgrowth endothelial progenitors isolated from peripheral blood of burned patients. , 2013, Burns : journal of the International Society for Burn Injuries.

[7]  Seungmin Lee,et al.  Early Vessel Destabilization Mediated by Angiopoietin-2 and Subsequent Vessel Maturation via Angiopoietin-1 Induce Functional Neovasculature after Ischemia , 2013, PloS one.

[8]  J. Bogaert,et al.  Placental growth factor increases regional myocardial blood flow and contractile function in chronic myocardial ischemia. , 2013, American journal of physiology. Heart and circulatory physiology.

[9]  S. Stringer,et al.  Age‐related impairment of endothelial progenitor cell migration correlates with structural alterations of heparan sulfate proteoglycans , 2013, Aging cell.

[10]  D. Stewart,et al.  Tie2-Dependent Neovascularization of the Ischemic Hindlimb Is Mediated by Angiopoietin-2 , 2012, PloS one.

[11]  P. Carmeliet,et al.  PlGF: a multitasking cytokine with disease-restricted activity. , 2012, Cold Spring Harbor perspectives in medicine.

[12]  C. Siu,et al.  Reversal of endothelial progenitor cell dysfunction in patients with type 2 diabetes using a conditioned medium of human embryonic stem cell‐derived endothelial cells , 2012, Diabetes/metabolism research and reviews.

[13]  J. Montaner,et al.  Mobilization, endothelial differentiation and functional capacity of endothelial progenitor cells after ischemic stroke. , 2010, Microvascular research.

[14]  Benoit Hendrickx,et al.  Integration of Blood Outgrowth Endothelial Cells in Dermal Fibroblast Sheets Promotes Full Thickness Wound Healing , 2010, Stem cells.

[15]  F. Van de Werf,et al.  Differential effects of progenitor cell populations on left ventricular remodeling and myocardial neovascularization after myocardial infarction. , 2010, Journal of the American College of Cardiology.

[16]  C. Boileau,et al.  Enhanced late-outgrowth circulating endothelial progenitor cell levels in rheumatoid arthritis and correlation with disease activity , 2010, Arthritis research & therapy.

[17]  M. Wolzt,et al.  Reduction of both number and proliferative activity of human endothelial progenitor cells in obesity , 2010, International Journal of Obesity.

[18]  W. Seeger,et al.  Evidence of dysfunction of endothelial progenitors in pulmonary arterial hypertension. , 2009, American journal of respiratory and critical care medicine.

[19]  M. Yoder.  Defining human endothelial progenitor cells , 2009, Journal of thrombosis and haemostasis : JTH.

[20]  G. Truskey,et al.  Comparison of endothelial cell phenotypic markers of late-outgrowth endothelial progenitor cells isolated from patients with coronary artery disease and healthy volunteers. , 2009, Tissue engineering. Part A.

[21]  R. Schnabel,et al.  Activated thrombin activatable fibrinolysis inhibitor levels are associated with the risk of cardiovascular death in patients with coronary artery disease: the AtheroGene study , 2009, Journal of thrombosis and haemostasis : JTH.

[22]  V. Rosti,et al.  Rapid and large increase of the frequency of circulating endothelial colony-forming cells (ECFCs) generating late outgrowth endothelial cells in patients with acute myocardial infarction. , 2009, Experimental hematology.

[23]  W. R. Taylor,et al.  Angiopoietin-2 Stimulates Blood Flow Recovery After Femoral Artery Occlusion by Inducing Inflammation and Arteriogenesis , 2008, Arteriosclerosis, thrombosis, and vascular biology.

[24]  P. Carmeliet,et al.  Beneficial effects of prolonged systemic administration of PlGF on late outcome of post‐ischaemic myocardial performance , 2008, The Journal of pathology.

[25]  K. Hirschi,et al.  Assessing identity, phenotype, and fate of endothelial progenitor cells. , 2008, Arteriosclerosis, thrombosis, and vascular biology.

[26]  Darby J. S. Thompson,et al.  Late outgrowth endothelial progenitor cells in patients with age-related macular degeneration. , 2008, Investigative ophthalmology & visual science.

[27]  J. Roh,et al.  Circulating Endothelial Progenitor Cells as a New Marker of Endothelial Dysfunction or Repair in Acute Stroke , 2008, Stroke.

[28]  A. Avogaro,et al.  Technical notes on endothelial progenitor cells: ways to escape from the knowledge plateau. , 2008, Atherosclerosis.

[29]  C. Heeschen,et al.  Selective functional exhaustion of hematopoietic progenitor cells in the bone marrow of patients with postinfarction heart failure. , 2007, Journal of the American College of Cardiology.

[30]  J. Prchal,et al.  Redefining endothelial progenitor cells via clonal analysis and hematopoietic stem/progenitor cell principals. , 2007, Blood.

[31]  P. Poole‐Wilson,et al.  Age-Dependent Impairment of Endothelial Progenitor Cells Is Corrected by Growth Hormone Mediated Increase of Insulin-Like Growth Factor-1 , 2007, Circulation research.

[32]  A. Avogaro,et al.  Number and Function of Endothelial Progenitor Cells as a Marker of Severity for Diabetic Vasculopathy , 2006, Arteriosclerosis, thrombosis, and vascular biology.

[33]  A. Jakobsson,et al.  Prediction of sampling depth and photon pathlength in laser Doppler flowmetry , 1993, Medical and Biological Engineering and Computing.

[34]  W. Schaper,et al.  Enhanced arteriogenesis in mice overexpressing erythropoietin , 2006, Cell and Tissue Research.

[35]  Stefanie Dimmeler,et al.  Impaired CXCR4 Signaling Contributes to the Reduced Neovascularization Capacity of Endothelial Progenitor Cells From Patients With Coronary Artery Disease , 2005, Circulation research.

[36]  G. Nickenig,et al.  Circulating endothelial progenitor cells and cardiovascular outcomes. , 2005, The New England journal of medicine.

[37]  W. Schaper,et al.  Preconditioning of arteriogenesis. , 2005, Cardiovascular research.

[38]  Matthias Heil,et al.  Pathophysiology of collateral development. , 2004, Coronary artery disease.

[39]  K. Pollok,et al.  Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood. , 2004, Blood.

[40]  Stefanie Dimmeler,et al.  Profoundly Reduced Neovascularization Capacity of Bone Marrow Mononuclear Cells Derived From Patients With Chronic Ischemic Heart Disease , 2004, Circulation.

[41]  C. Dehio,et al.  Coordinated activation of VEGFR-1 and VEGFR-2 is a potent arteriogenic stimulus leading to enhancement of regional perfusion. , 2004, Cardiovascular research.

[42]  A. Quyyumi,et al.  Circulating endothelial progenitor cells, vascular function, and cardiovascular risk. , 2003, The New England journal of medicine.

[43]  W. Schaper,et al.  Factors Regulating Arteriogenesis , 2003, Arteriosclerosis, thrombosis, and vascular biology.

[44]  P. Carmeliet,et al.  VEGFR-1–Selective VEGF Homologue PlGF Is Arteriogenic: Evidence for a Monocyte-Mediated Mechanism , 2003, Circulation research.

[45]  A. Luttun,et al.  Revascularization of ischemic tissues by PlGF treatment, and inhibition of tumor angiogenesis, arthritis and atherosclerosis by anti-Flt1 , 2002, Nature Medicine.

[46]  M. Pasic,et al.  The problem of chronic refractory angina; report from the ESC Joint Study Group on the Treatment of Refractory Angina. , 2002, European heart journal.

[47]  S. Fichtlscherer,et al.  Number and Migratory Activity of Circulating Endothelial Progenitor Cells Inversely Correlate With Risk Factors for Coronary Artery Disease , 2001, Circulation research.

[48]  H. Augustin,et al.  Tensional forces in fibrillar extracellular matrices control directional capillary sprouting. , 1999, Journal of cell science.