title: β Adrenergic Receptor Kinase C-Terminal Peptide Gene-Therapy Improves β2-Adrenergic Receptor-Dependent Neoangiogenesis after Hindlimb Ischemia.
abstract: After hindlimb ischemia (HI), increased catecholamine levels within the ischemic muscle can cause dysregulation of β2-adrenergic receptor (β2AR) signaling, leading to reduced revascularization. Indeed, in vivo β2AR overexpression via gene therapy enhances angiogenesis in a rat model of HI. G protein-coupled receptor kinase 2 (GRK2) is a key regulator of βAR signaling, and β adrenergic receptor kinase C-terminal peptide (βARKct), a peptide inhibitor of GRK2, has been shown to prevent βAR down-regulation and to protect cardiac myocytes and stem cells from ischemic injury through restoration of β2AR protective signaling (i.e., protein kinase B/endothelial nitric oxide synthase). Herein, we tested the potential therapeutic effects of adenoviral-mediated βARKct gene transfer in an experimental model of HI and its effects on βAR signaling and on endothelial cell (EC) function in vitro. Accordingly, in this study, we surgically induced HI in rats by femoral artery resection (FAR). Fifteen days of ischemia resulted in significant βAR down-regulation that was paralleled by an approximately 2-fold increase in GRK2 levels in the ischemic muscle. Importantly, in vivo gene transfer of the βARKct in the hindlimb of rats at the time of FAR resulted in a marked improvement of hindlimb perfusion, with increased capillary and βAR density in the ischemic muscle, compared with control groups. The effect of βARKct expression was also assessed in vitro in cultured ECs. Interestingly, ECs expressing the βARKct fenoterol, a β2AR-agonist, induced enhanced β2AR proangiogenic signaling and increased EC function. Our results suggest that βARKct gene therapy and subsequent GRK2 inhibition promotes angiogenesis in a model of HI by preventing ischemia-induced β2AR down-regulation.
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.).
authors.forename: Alessandro
authors.initials: A
authors.lastname: Cannavo
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.).
authors.forename: Daniela
authors.initials: D
authors.lastname: Liccardo
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.).
authors.forename: Anastasios
authors.initials: A
authors.lastname: Lymperopoulos
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.).
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Gambino
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.).
authors.forename: Maria Loreta
authors.initials: ML
authors.lastname: D'Amico
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.).
authors.forename: Franco
authors.initials: F
authors.lastname: Rengo
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.).
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Koch
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.) dleosco@unina.it.
authors.forename: Dario
authors.initials: D
authors.lastname: Leosco
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.).
authors.forename: Nicola
authors.initials: N
authors.lastname: Ferrara
authors.affiliation: Division of Geriatrics, Department of Translational Medical Sciences, Federico II University of Naples, Italy (A.C., D.Li., G.G., M.L.D.A., D.Le., N.F., G.R.); Center for Translational Medicine, Temple University, Philadelphia, Pennsylvania (A.C., D.Li., W.J.K.); Department of Pharmaceutical Sciences, Nova Southeastern University College of Pharmacy, Fort Lauderdale, Florida (A.L.); Salvatore Maugeri Foundation, IRCCS, Scientific Institute of Telese Terme BN, Italy (F.R., G.R.).
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Rengo
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D015316:Genetic Therapy / Q000639:trends*; D006614:Hindlimb / Q000098:blood supply*; D007511:Ischemia / Q000235:genetics* / Q000628:therapy; D008297:Male; D009389:Neovascularization, Pathologic / Q000235:genetics* / Q000628:therapy; D010446:Peptide Fragments / Q000008:administration & dosage / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics* / Q000378:metabolism; D051552:beta-Adrenergic Receptor Kinases / Q000008:administration & dosage / Q000235:genetics*
chemical_list: D010446:Peptide Fragments; D018343:Receptors, Adrenergic, beta-2; D051552:beta-Adrenergic Receptor Kinases