Identification of Small‐Molecule PHD2 Zinc Finger Inhibitors that Activate Hypoxia Inducible Factor

Arsenault, Patrick R.; Song, Daisheng; Bergkamp, Marian; Ravaschiere, Andrew M.; Navalsky, Bradleigh E.; Lieberman, Paul M.; Lee, Frank S.

doi:10.1002/cbic.201600493

Cited by 6 publications

(6 citation statements)

References 36 publications

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“…The interaction with pVHL requires the hydroxylation of specific proline residues of AKT1 [21] (table 1). AKT1 hydroxylation is mediated by PHD2, which in turn is the main regulator of HIF-1α hydroxylation [23,[50][51][52]. The interaction promotes AKT1 functional inhibition, while, similarly to what was observed for PKCδ, it seems to have no effect on AKT1 degradation.…”

Section: Von Hippel-lindau Protein and The Regulation Of Kinases Activitymentioning

confidence: 70%

The pVHL neglected functions, a tale of hypoxia-dependent and -independent regulations in cancer

2020

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The von Hippel–Lindau protein (pVHL) is a tumour suppressor mainly known for its role as master regulator of hypoxia-inducible factor (HIF) activity. Functional inactivation of pVHL is causative of the von Hippel–Lindau disease, an inherited predisposition to develop different cancers. Due to its impact on human health, pVHL has been widely studied in the last few decades. However, investigations mostly focus on its role in degrading HIFs, whereas alternative pVHL protein–protein interactions and functions are insistently surfacing in the literature. In this review, we analyse these almost neglected functions by dissecting specific conditions in which pVHL is proposed to have differential roles in promoting cancer. We reviewed its role in regulating phosphorylation as a number of works suggest pVHL to act as an inhibitor by either degrading or promoting downregulation of specific kinases. Further, we summarize hypoxia-dependent and -independent pVHL interactions with multiple protein partners and discuss their implications in tumorigenesis.

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Section: Von Hippel-lindau Protein and The Regulation Of Kinases Activitymentioning

confidence: 70%

The pVHL neglected functions, a tale of hypoxia-dependent and -independent regulations in cancer

2020

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“…S6E). PHD2 contains a highly conserved myeloid nervy deformed epidermal autoregulatory factor 1 homolog (MYND)–type zinc finger ( 28 , 29 ) with two conserved CXXC (two cysteines separated by two other residues) domains C21/24 and C33/36; the two mutations that affect PHD2 activity fall within one or the other of these conserved domains. To determine whether the identified cysteine residues within PHD2 were important for association with HIF-1a, we assessed the interaction of mutant or WT PHD2 with full-length glutathione S -transferase (GST)–HIF-1α.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to its C-terminal PHD domain, PHD2 also has an N-terminal, evolutionary-conserved, MYND-type zinc finger, a feature that is absent in both PHD1 and PHD3 ( 28 , 29 ). The zinc finger of PHD2 is essential for efficient hydroxylation of HIF-1α ( 11 , 28 , 29 ). Supporting the essentiality of the zinc finger, we found that H 2 S persulfidates PHD2 at five different cysteine residues, two of which are within the zinc finger motif.…”

Section: Discussionmentioning

confidence: 99%

Cystathione β-synthase regulates HIF-1α stability through persulfidation of PHD2

et al. 2020

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The stringent expression of the hypoxia inducible factor-1α (HIF-1α) is critical to a variety of pathophysiological conditions. We reveal that, in normoxia, enzymatic action of cystathionine β-synthase (CBS) produces H2S, which persulfidates prolyl hydroxylase 2 (PHD2) at residues Cys21 and Cys33 (zinc finger motif), augmenting prolyl hydroxylase activity. Depleting endogenous H2S either by hypoxia or by inhibiting CBS via chemical or genetic means reduces persulfidation of PHD2 and inhibits activity, preventing hydroxylation of HIF-1α, resulting in stabilization. Our in vitro findings are further supported by the depletion of CBS in the zebrafish model that exhibits axis defects and abnormal intersegmental vessels. Exogenous H2S supplementation rescues both in vitro and in vivo phenotypes. We have identified the persulfidated residues and defined their functional significance in regulating the activity of PHD2 via point mutations. Thus, the CBS/H2S/PHD2 axis may provide therapeutic opportunities for pathologies associated with HIF-1α dysregulation in chronic diseases.

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“…2C). We have previously generated the corresponding control mouse line in which the coding sequence of murine Phd2 exon 1 is replaced by that of wild-type human PHD2 exon 1 (hereafter referred to as Phd2 WT ) (24). There was no difference in litter sizes between the two genotypes (5.19 ± 1.38 [SD] for Phd2 WT/WT × Phd2 WT/WT crosses vs. 5.00 ± 1.93 [SD] for Phd2 Tib/Tib × Phd2 Tib/Tib crosses; n = 16 litters for each; P = 0.75).…”

Section: Mutations In the Vicinity Of Asp-4 And Cys-127 Of Phd2 Impairmentioning

confidence: 99%

Tibetan PHD2 , an allele with loss-of-function properties

Song

Navalsky

Guan

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Tibetans have adapted to the chronic hypoxia of high altitude and display a distinctive suite of physiologic adaptations, including augmented hypoxic ventilatory response and resistance to pulmonary hypertension. Genome-wide studies have consistently identified compelling genetic signatures of natural selection in two genes of the Hypoxia Inducible Factor pathway,PHD2andHIF2A. The product of the former induces the degradation of the product of the latter. Key issues regarding TibetanPHD2are whether it is a gain-of-function or loss-of-function allele, and how it might contribute to high-altitude adaptation. Tibetan PHD2 possesses two amino acid changes, D4E and C127S. We previously showed that in vitro, Tibetan PHD2 is defective in its interaction with p23, a cochaperone of the HSP90 pathway, and we proposed that TibetanPHD2is a loss-of-function allele. Here, we report that additional PHD2 mutations at or near Asp-4 or Cys-127 impair interaction with p23 in vitro. We find that mice with the TibetanPhd2allele display augmented hypoxic ventilatory response, supporting this loss-of-function proposal. This is phenocopied by mice with a mutation inp23that abrogates the PHD2:p23 interaction.Hif2ahaploinsufficiency, but not the TibetanPhd2allele, ameliorates hypoxia-induced increases in right ventricular systolic pressure. The TibetanPhd2allele is not associated with hemoglobin levels in mice. We propose that Tibetans possess genetic alterations that both activate and inhibit selective outputs of the HIF pathway to facilitate successful adaptation to the chronic hypoxia of high altitude.

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Identification of Small‐Molecule PHD2 Zinc Finger Inhibitors that Activate Hypoxia Inducible Factor

Cited by 6 publications

References 36 publications

The pVHL neglected functions, a tale of hypoxia-dependent and -independent regulations in cancer

The pVHL neglected functions, a tale of hypoxia-dependent and -independent regulations in cancer

Cystathione β-synthase regulates HIF-1α stability through persulfidation of PHD2

Tibetan PHD2 , an allele with loss-of-function properties

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