title: Identification of Small-Molecule PHD2 Zinc Finger Inhibitors that Activate Hypoxia Inducible Factor.
abstract: The prolyl hydroxylase domain (PHD) protein:hypoxia inducible factor (HIF) pathway is the main pathway by which changes in oxygen concentration are transduced to changes in gene expression. In mammals, there are three PHD paralogues, and PHD2 has emerged as a particularly critical one for regulating HIF target genes such as erythropoietin (EPO), which controls red cell mass and hematocrit. PHD2 is distinctive among the three PHDs in that it contains an N-terminal MYND-type zinc finger. We have proposed that this zinc finger binds a Pro-Xaa-Leu-Glu (PXLE) motif found in proteins of the HSP90 pathway to facilitate HIF-α hydroxylation. Targeting this motif could provide a means of specifically inhibiting this PHD isoform. Here, we screened a library of chemical compounds for their capacity to inhibit the zinc finger of PHD2. We identified compounds that, in vitro, can inhibit PHD2 binding to a PXLE-containing peptide and induce activation of HIF. Injection of one of these compounds into mice induces an increase in hematocrit. This study offers proof of principle that inhibition of the zinc finger of PHD2 can provide a means of selectively targeting PHD2 to activate the HIF pathway.
authors: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, 605 Stellar Chance Labs, 422 Curie Blvd, Philadelphia, PA, 19104, USA.
authors: Patrick R
authors: PR
authors: Arsenault
authors: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, 605 Stellar Chance Labs, 422 Curie Blvd, Philadelphia, PA, 19104, USA.
authors: Daisheng
authors: D
authors: Song
authors: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, 605 Stellar Chance Labs, 422 Curie Blvd, Philadelphia, PA, 19104, USA.
authors: Marian
authors: M
authors: Bergkamp
authors: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, 605 Stellar Chance Labs, 422 Curie Blvd, Philadelphia, PA, 19104, USA.
authors: Andrew M
authors: AM
authors: Ravaschiere
authors: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, 605 Stellar Chance Labs, 422 Curie Blvd, Philadelphia, PA, 19104, USA.
authors: Bradleigh E
authors: BE
authors: Navalsky
authors: The Wistar Institute, 3601 Spruce St., Philadelphia, PA, 19104, USA.
authors: Paul M
authors: PM
authors: Lieberman
authors: Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, 605 Stellar Chance Labs, 422 Curie Blvd, Philadelphia, PA, 19104, USA.
authors: Frank S
authors: FS
authors: Lee
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics; D020134:Catalytic Domain; D004921:Erythropoietin / Q000097:blood / Q000235:genetics / Q000378:metabolism; D055879:Gene Knock-In Techniques; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism; D006400:Hematocrit; D006801:Humans; D006900:Hydroxylation; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D054852:Small Molecule Libraries / Q000737:chemistry / Q000378:metabolism; D016335:Zinc Fingers
chemical_list: D018841:HSP90 Heat-Shock Proteins; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D004921:Erythropoietin; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases
keywords: EGLN1; PHD2; drug discovery; high-throughput screening; hypoxia-inducible factor