title: Synergistic effect of HIF-1alpha gene therapy and HIF-1-activated bone marrow-derived angiogenic cells in a mouse model of limb ischemia.
abstract: Ischemia induces the production of angiogenic cytokines and the homing of bone-marrow-derived angiogenic cells (BMDACs), but these adaptive responses become impaired with aging because of reduced expression of hypoxia-inducible factor (HIF)-1alpha. In this study, we analyzed the effect of augmenting HIF-1alpha levels in ischemic limb by intramuscular injection of AdCA5, an adenovirus encoding a constitutively active form of HIF-1alpha, and intravenous administration of BMDACs that were cultured in the presence of the prolyl-4-hydroxylase inhibitor dimethyloxalylglycine (DMOG) to induce HIF-1 expression. The combined therapy increased perfusion, motor function, and limb salvage in old mice subjected to femoral artery ligation. Homing of BMDACs to the ischemic limb was dramatically enhanced by intramuscular AdCA5 administration. DMOG treatment of BMDACs increased cell surface expression of beta(2) integrins, which mediated increased adherence of BMDACs to endothelial cells. The effect of DMOG was abolished by coadministration of the HIF-1 inhibitor digoxin or by preincubation with a beta(2) integrin-blocking antibody. Transduction of BMDACs with lentivirus LvCA5 induced effects similar to DMOG treatment. Thus, HIF-1alpha gene therapy increases homing of BMDACs to ischemic muscle, whereas HIF-1 induction in BMDACs enhances their adhesion to vascular endothelium, leading to synergistic effects of combined therapy on tissue perfusion.
authors: Vascular Program, Institute for Cell Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors: Sergio
authors: S
authors: Rey
authors: KangAe
authors: K
authors: Lee
authors: C Joanne
authors: CJ
authors: Wang
authors: Kshitiz
authors: K
authors: Gupta
authors: Shaoping
authors: S
authors: Chen
authors: Alexandra
authors: A
authors: McMillan
authors: Nupura
authors: N
authors: Bhise
authors: Andre
authors: A
authors: Levchenko
authors: Gregg L
authors: GL
authors: Semenza
mesh_terms: D000256:Adenoviridae; D000367:Age Factors; D000600:Amino Acids, Dicarboxylic / Q000494:pharmacology; D043925:Angiogenesis Inducing Agents / Q000378:metabolism; D000818:Animals; D016026:Bone Marrow Transplantation; D002448:Cell Adhesion / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D005263:Femoral Artery / Q000601:surgery; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D006614:Hindlimb / Q000473:pathology*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D007273:Injections, Intramuscular; D007511:Ischemia / Q000628:therapy*; D008026:Ligation; D051379:Mice; D011393:Procollagen-Proline Dioxygenase / Q000037:antagonists & inhibitors; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000600:Amino Acids, Dicarboxylic; D043925:Angiogenesis Inducing Agents; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D011393:Procollagen-Proline Dioxygenase; C040947:oxalylglycine