Neovascularization Potential of Blood Outgrowth Endothelial Cells From Patients With Stable Ischemic Heart Failure Is Preserved.
Dieter Dauwe, Beatriz Pelacho, Arief Wibowo, Ann-Sophie Walravens, Kristoff Verdonck, Hilde Gillijns, Ellen Caluwe, Peter Pokreisz, Nick van Gastel, Geert Carmeliet, Maarten Depypere, Frederik Maes, Nina Vanden Driessche, Walter Droogne, Johan Van Cleemput, Johan Vanhaecke, Felipe Prosper, Catherine Verfaillie, Aernout Luttun, Stefan Janssens
Author Information
Dieter Dauwe: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium dieter.dauwe@med.kuleuven.be stefan.janssens@med.kuleuven.be.
Beatriz Pelacho: Cell Therapy Department, Center for Applied Medicine Research, Clinica Universidad de Navarra, University of Navarra, Pamplona, Spain.
Arief Wibowo: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
Ann-Sophie Walravens: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
Kristoff Verdonck: Center for Molecular and Vascular Biology, KU Leuven, Leuven, Belgium.
Hilde Gillijns: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
Ellen Caluwe: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
Peter Pokreisz: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
Nick van Gastel: Department of Clinical and Experimental Endocrinology, KU Leuven, Leuven, Belgium.
Geert Carmeliet: Department of Clinical and Experimental Endocrinology, KU Leuven, Leuven, Belgium.
Maarten Depypere: Department of Electrical Engineering, Center for the Processing of Speech and Images, KU Leuven, Leuven, Belgium.
Frederik Maes: Department of Electrical Engineering, Center for the Processing of Speech and Images, KU Leuven, Leuven, Belgium.
Nina Vanden Driessche: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
Walter Droogne: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
Johan Van Cleemput: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
Johan Vanhaecke: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium.
Felipe Prosper: Cell Therapy Department, Center for Applied Medicine Research, Clinica Universidad de Navarra, University of Navarra, Pamplona, Spain Hematology Department, Clinica Universidad de Navarra, University of Navarra, Pamplona, Spain.
Catherine Verfaillie: Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven, Leuven, Belgium.
Aernout Luttun: Center for Molecular and Vascular Biology, KU Leuven, Leuven, Belgium.
Stefan Janssens: Department of Cardiovascular Sciences, Clinical Cardiology, KU Leuven, Leuven, Belgium dieter.dauwe@med.kuleuven.be stefan.janssens@med.kuleuven.be.
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/100×10(6) 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/100×10(6) MNCs in ACON (n=32; age: 60±1 years) and 2.6±0.4 colonies/100×10(6) 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±0.3×10(5) pg/10(6) ICMP-BOECs) and placental growth factor (5.8±1.5×10(3) pg/10(6) 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.
