Activation of the hypoxia-inducible factor-pathway and stimulation of angiogenesis by application of prolyl hydroxylase inhibitors.
Hypoxia-inducible transcription factors (HIF) mediate complex adaptations to reduced oxygen supply, including neoangiogenesis. Regulation of HIF occurs mainly through oxygen-dependent destruction of its alpha subunit. In the presence of oxygen, two HIFalpha prolyl residues undergo enzymatic hydroxylation, which is required for its proteasomal degradation. We therefore tested whether pharmacological activation of HIFalpha by hydroxylase inhibitors may provide a novel therapeutic strategy for the treatment of ischemic diseases. Three distinct prolyl 4-hydroxylase inhibitors-l-mimosine (L-Mim), ethyl 3,4-dihydroxybenzoate (3,4-DHB), and 6-chlor-3-hydroxychinolin-2-carbonic acid-N-carboxymethylamid (S956711)-demonstrated similar effects to hypoxia (0.5% O2) by inducing HIFalpha protein in human and rodent cells. L-Mim, S956711, and, less effectively, 3,4-DHB also induced HIF target genes in cultured cells, including glucose transporter 1 and vascular endothelial growth factor, as well as HIF-dependent reporter gene expression. Systemic administration of L-Mim and S956711 in rats led to HIFalpha induction in the kidney. In a sponge model for angiogenesis, repeated local injection of the inhibitors strongly increased invasion of highly vascularized tissue into the sponge centers. In conclusion, structurally distinct inhibitors of prolyl hydroxylation are capable of inducing HIFalpha and HIF target genes in vitro and in vivo and induce adaptive responses to hypoxia, including angiogenesis.
Warnecke
Christina
C
Department of Nephrology and Medical Intensive Care, Charité, Campus Virchow Clinic, Augustenburger Platz 1, 13353 Berlin, Germany.
Griethe
Wanja
W
Weidemann
Alexander
A
Jürgensen
Jan Steffen
JS
Willam
Carsten
C
Bachmann
Sebastian
S
Ivashchenko
Yuri
Y
Wagner
Ingrid
I
Frei
Ulrich
U
Wiesener
Michael
M
Eckardt
Kai-Uwe
KU
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity; D006801:Humans; D062385:Hydroxybenzoates / Q000494:pharmacology / Q000633:toxicity; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008025:Ligases / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008898:Mimosine / Q000494:pharmacology / Q000633:toxicity; D008954:Models, Biological; D018919:Neovascularization, Physiologic; D011393:Procollagen-Proline Dioxygenase / Q000037:antagonists & inhibitors*; D011804:Quinolines / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation; D025521:Tumor Suppressor Proteins; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein
C474763:6-chlor-3-hydroxychinolin-2-carbonic acid N-carboxymethylamid; D004791:Enzyme Inhibitors; C497442:HIF1A protein, human; D062385:Hydroxybenzoates; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D011804:Quinolines; D012333:RNA, Messenger; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C018063:ethyl protocatechuate; D008898:Mimosine; D008156:Luciferases; D011393:Procollagen-Proline Dioxygenase; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D008025:Ligases; C497446:VHL protein, human