title: Hypoxia inducible factor stabilization improves defective ischemia-induced angiogenesis in a rodent model of chronic kidney disease.
abstract: Chronic kidney disease (CKD) is associated with increased risk and worse prognosis of cardiovascular disease, including peripheral artery disease. An impaired angiogenic response to ischemia may contribute to poor outcomes of peripheral artery disease in patients with CKD. Hypoxia inducible factors (HIF) are master regulators of angiogenesis and therefore represent a promising target for therapeutic intervention. To test this we induced hind-limb ischemia in rats with CKD caused by 5/6 nephrectomy and administered two different treatments known to stabilize HIF protein in vivo: carbon monoxide and a pharmacological inhibitor of prolyl hydroxylation 2-(1-chloro-4- hydroxyisoquinoline-3-carboxamido) acetate (ICA). Expression levels of pro-angiogenic HIF target genes (Vegf, Vegf-r1, Vegf-r2, Ho-1) were measured by qRT-PCR. Capillary density was measured by CD31 immunofluorescence staining and HIF expression was evaluated by immunohistochemistry. Capillary density in ischemic skeletal muscle was significantly lower in CKD animals compared to sham controls. Rats with CKD showed significantly lower expression of HIF and all measured pro-angiogenic HIF target genes, including VEGF. Both HIF stabilizing treatments rescued HIF target gene expression in animals with CKD and led to significantly higher ischemia-induced capillary sprouting compared to untreated controls. ICA was effective regardless of whether it was administered before or after induction of ischemia and led to a HIF expression in skeletal muscle. Thus, impaired ischemia-induced angiogenesis in rats with CKD can be improved by HIF stabilization, even if started after onset of ischemia.
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany; Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, California, USA; VA Palo Alto Health Care System, Palo Alto, California, USA; Heart Center, Georg-August-University Göttingen, Göttingen, Germany; German Center for Cardiovascular Research (DZHK), Göttingen, Germany.
authors.forename: Isabel N
authors.initials: IN
authors.lastname: Schellinger
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Nada
authors.initials: N
authors.lastname: Cordasic
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Julian
authors.initials: J
authors.lastname: Panesar
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Björn
authors.initials: B
authors.lastname: Buchholz
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Jacobi
authors.affiliation: Department of Pediatrics and Adolescent Medicine, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Hartner
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Klanke
authors.affiliation: Heart Center, Georg-August-University Göttingen, Göttingen, Germany.
authors.forename: Joanna
authors.initials: J
authors.lastname: Jakubiczka-Smorag
authors.affiliation: Department of Chemistry and Pharmacy, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Nicolai
authors.initials: N
authors.lastname: Burzlaff
authors.affiliation: Department of Chemistry and Pharmacy, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Heinze
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Warnecke
authors.affiliation: Heart Center, Georg-August-University Göttingen, Göttingen, Germany; German Center for Cardiovascular Research (DZHK), Göttingen, Germany.
authors.forename: Uwe
authors.initials: U
authors.lastname: Raaz
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Willam
authors.affiliation: Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, California, USA; VA Palo Alto Health Care System, Palo Alto, California, USA; Cardiovascular Institute, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Philip S
authors.initials: PS
authors.lastname: Tsao
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Kai-Uwe
authors.initials: KU
authors.lastname: Eckardt
authors.affiliation: Division of Nephropathology, University of Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Amann
authors.affiliation: Department of Nephrology and Hypertension, University of Erlangen-Nürnberg, Erlangen, Germany. Electronic address: karl.hilgers@uk-erlangen.de.
authors.forename: Karl F
authors.initials: KF
authors.lastname: Hilgers
mesh_terms: D000818:Animals; D002196:Capillaries / Q000187:drug effects* / Q000378:metabolism / Q000503:physiopathology; D002248:Carbon Monoxide / Q000494:pharmacology*; D002460:Cell Line; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006419:Heme Oxygenase (Decyclizing) / Q000235:genetics / Q000378:metabolism; D006614:Hindlimb; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D007511:Ischemia / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D007546:Isoquinolines / Q000494:pharmacology*; D008297:Male; D018482:Muscle, Skeletal / Q000098:blood supply*; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D055550:Protein Stability; D017207:Rats, Sprague-Dawley; D051436:Renal Insufficiency, Chronic / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D015398:Signal Transduction / Q000187:drug effects*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
chemical_list: C000608512:2-(1-chloro-4-hydroxyisoquinoline-3-carboxamido)acetic acid; C497442:HIF1A protein, human; C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007546:Isoquinolines; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat; D002248:Carbon Monoxide; D006419:Heme Oxygenase (Decyclizing); C491523:Hmox1 protein, rat; C501164:Flt1 protein, rat; C000602029:Kdr protein, rat; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2; D005998:Glycine
keywords: cardiovascular disease; chronic kidney disease; hypoxia