title: Endothelin-converting enzyme is a plausible target gene for hypoxia-inducible factor.
abstract: Renal endothelin-converting enzyme (ECE)-1 is induced in experimental diabetes and following radiocontrast administration, conditions characterized by renal hypoxia, hypoxia-inducible factor (HIF) stabilization, and enhanced endothelin synthesis. Here we tested whether ECE-1 might be a HIF-target gene in vitro and in vivo. ECE-1 transcription and expression increased in cultured vascular endothelial and proximal tubular cell lines, subject to hypoxia, to mimosine or cobalt chloride. These interventions are known to stabilize HIF signaling by inhibition of HIF-prolyl hydroxylases. In rats, HIF-prolyl-hydroxylase inhibition by mimosine or FG-4497 increased HIF-1α immunostaining in renal tubules, principally in distal nephron segments. This was associated with markedly enhanced ECE-1 protein expression, predominantly in the renal medulla. A progressive and dramatic increase in ECE-1 immunostaining over time, in parallel with enhanced HIF expression, was also noted in conditional von Hippel-Lindau knockout mice. Since HIF and STAT3 are cross-stimulated, we triggered HIF expression by STAT3 activation in mice, transfected by or injected with a chimeric IL-6/IL-6-receptor protein, and found a similar pattern of enhanced ECE-1 expression. Chromatin immunoprecipitation sequence (ChIP-seq) and PCR analysis in hypoxic endothelial cells identified HIF binding at the ECE-1 promoter and intron regions. Thus, our findings suggest that ECE-1 may be a novel HIF-target gene.
authors.affiliation: Institute of Endocrinology, Diabetes & Metabolism, Rambam Medical Center & RB Rappaport Faculty of Medicine-Technion, Haifa, Israel.
authors.forename: Mogher
authors.initials: M
authors.lastname: Khamaisi
authors.affiliation: Institute of Endocrinology, Diabetes & Metabolism, Rambam Medical Center & RB Rappaport Faculty of Medicine-Technion, Haifa, Israel.
authors.forename: Hala
authors.initials: H
authors.lastname: Toukan
authors.affiliation: Goldyne Savad Institute of Gene Therapy, Hebrew University, Hadassah Hospitals, Jerusalem, Israel.
authors.forename: Jonathan H
authors.initials: JH
authors.lastname: Axelrod
authors.affiliation: Department of Nephrology, Charité, Berlin, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Rosenberger
authors.affiliation: Department of Medicine, Hebrew University, Hadassah Hospitals, Jerusalem, Israel.
authors.forename: Galia
authors.initials: G
authors.lastname: Skarzinski
authors.affiliation: Department of Medicine, Hebrew University, Hadassah Hospitals, Jerusalem, Israel.
authors.forename: Ahuva
authors.initials: A
authors.lastname: Shina
authors.affiliation: Department of Animal Sciences, The Robert H Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel.
authors.forename: Rina
authors.initials: R
authors.lastname: Meidan
authors.affiliation: Department of Nephrology, Tenon Hospital, University Pierre et Marie Curie, Paris, France.
authors.forename: Robert
authors.initials: R
authors.lastname: Koesters
authors.affiliation: Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Seymour
authors.initials: S
authors.lastname: Rosen
authors.affiliation: FibroGen, San Francisco, California, USA.
authors.forename: Gail
authors.initials: G
authors.lastname: Walkinshaw
authors.affiliation: Divisions of Nephrology and Endocrinology, University of Tokyo School of Medicine, Tokyo, Japan.
authors.forename: Imari
authors.initials: I
authors.lastname: Mimura
authors.affiliation: Divisions of Nephrology and Endocrinology, University of Tokyo School of Medicine, Tokyo, Japan.
authors.forename: Masaomi
authors.initials: M
authors.lastname: Nangaku
authors.affiliation: Department of Medicine, Hebrew University, Hadassah Hospitals, Jerusalem, Israel.
authors.forename: Samuel N
authors.initials: SN
authors.lastname: Heyman
mesh_terms: D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D015687:Cell Hypoxia / Q000502:physiology; D002478:Cells, Cultured; D003035:Cobalt / Q000494:pharmacology; D049308:Dioxygenases / Q000037:antagonists & inhibitors; D000072396:Endothelin-Converting Enzymes; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D007438:Introns; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000378:metabolism; D008297:Male; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D008898:Mimosine / Q000494:pharmacology; D020411:Oligonucleotide Array Sequence Analysis; D064800:Prolyl-Hydroxylase Inhibitors / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D051381:Rats; D017207:Rats, Sprague-Dawley; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014158:Transcription, Genetic; D006623:von Hippel-Lindau Disease / Q000235:genetics / Q000378:metabolism
chemical_list: C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D064800:Prolyl-Hydroxylase Inhibitors; D050796:STAT3 Transcription Factor; C494087:Stat3 protein, mouse; D003035:Cobalt; D008898:Mimosine; D049308:Dioxygenases; C552717:HIF prolyl hydroxylase, rat; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; C000606923:ECE1 protein, human; C000606925:Ece1 protein, mouse; D000072396:Endothelin-Converting Enzymes; C018021:cobaltous chloride