Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition

Julianty Frost, Carles Galdeano, Pedro Soares, Morgan S. Gadd, Katarzyna M. Grzes, Lucy Ellis, Ola Epemolu, Satoko Shimamura, Marcus Bantscheff, Paola Grandi, Kevin D. Read, Doreen A. Cantrell, Sonia Rocha, Alessio Ciulli (Lead / Corresponding author)

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Abstract

Chemical strategies to using small molecules to stimulate hypoxia inducible factors (HIFs) activity and trigger a hypoxic response under normoxic conditions, such as iron chelators and inhibitors of prolyl hydroxylase domain (PHD) enzymes, have broad-spectrum activities and off-target effects. Here we disclose VH298, a potent VHL inhibitor that stabilizes HIF-α and elicits a hypoxic response via a different mechanism, that is the blockade of the VHL:HIF-α protein-protein interaction downstream of HIF-α hydroxylation by PHD enzymes. We show that VH298 engages with high affinity and specificity with VHL as its only major cellular target, leading to selective on-target accumulation of hydroxylated HIF-α in a concentration- and time-dependent fashion in different cell lines, with subsequent upregulation of HIF-target genes at both mRNA and protein levels. VH298 represents a high-quality chemical probe of the HIF signaling cascade and an attractive starting point to the development of potential new therapeutics targeting hypoxia signaling.
Original languageEnglish
Article number13312
Pages (from-to)1-12
Number of pages12
JournalNature Communications
Volume7
DOIs
Publication statusPublished - 4 Nov 2016

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Hydroxylation
hypoxia
probes
Prolyl-Hydroxylase Inhibitors
Prolyl Hydroxylases
Proteins
Enzymes
Chelating Agents
Iron
Genes
proteins
Cells
inhibitors
enzymes
Messenger RNA
Molecules
Hypoxia
cultured cells
genes
affinity

Cite this

Frost, Julianty ; Galdeano, Carles ; Soares, Pedro ; Gadd, Morgan S. ; Grzes, Katarzyna M. ; Ellis, Lucy ; Epemolu, Ola ; Shimamura, Satoko ; Bantscheff, Marcus ; Grandi, Paola ; Read, Kevin D. ; Cantrell, Doreen A. ; Rocha, Sonia ; Ciulli, Alessio. / Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition. In: Nature Communications. 2016 ; Vol. 7. pp. 1-12.
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title = "Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition",
abstract = "Chemical strategies to using small molecules to stimulate hypoxia inducible factors (HIFs) activity and trigger a hypoxic response under normoxic conditions, such as iron chelators and inhibitors of prolyl hydroxylase domain (PHD) enzymes, have broad-spectrum activities and off-target effects. Here we disclose VH298, a potent VHL inhibitor that stabilizes HIF-α and elicits a hypoxic response via a different mechanism, that is the blockade of the VHL:HIF-α protein-protein interaction downstream of HIF-α hydroxylation by PHD enzymes. We show that VH298 engages with high affinity and specificity with VHL as its only major cellular target, leading to selective on-target accumulation of hydroxylated HIF-α in a concentration- and time-dependent fashion in different cell lines, with subsequent upregulation of HIF-target genes at both mRNA and protein levels. VH298 represents a high-quality chemical probe of the HIF signaling cascade and an attractive starting point to the development of potential new therapeutics targeting hypoxia signaling.",
author = "Julianty Frost and Carles Galdeano and Pedro Soares and Gadd, {Morgan S.} and Grzes, {Katarzyna M.} and Lucy Ellis and Ola Epemolu and Satoko Shimamura and Marcus Bantscheff and Paola Grandi and Read, {Kevin D.} and Cantrell, {Doreen A.} and Sonia Rocha and Alessio Ciulli",
note = "This work was supported by the European Research Council ERC-2012-StG-311460 DrugE3CRLs (Starting Grant to A.C.), the UK Biotechnology and Biological Sciences Research Council BBSRC BB/G023123/2 (David Phillips Fellowship to A.C.), the European Commission PIEF-GA-2012-328030 (Marie-Curie Intra-European Fellowship to C.G.), the Wellcome Trust (PhD Studentship 102398/Z/13/Z to J.F., and strategic awards 100476/Z/12/Z for biophysics and drug discovery and 094090/Z/10/Z for structural biology and X-ray crystallography to BCDD) and the Fundac¸a˜o para a Cieˆncia e Tecnologia FCT SFRH/BD/101598/2014 (PhD Studentship to P.S.). S.R. is funded by a Cancer Research UK Senior fellowship C99667/A12918 with support of a Wellcome Trust strategic award 097945/B/11/Z.",
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Frost, J, Galdeano, C, Soares, P, Gadd, MS, Grzes, KM, Ellis, L, Epemolu, O, Shimamura, S, Bantscheff, M, Grandi, P, Read, KD, Cantrell, DA, Rocha, S & Ciulli, A 2016, 'Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition', Nature Communications, vol. 7, 13312, pp. 1-12. https://doi.org/10.1038/ncomms13312

Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition. / Frost, Julianty; Galdeano, Carles; Soares, Pedro; Gadd, Morgan S.; Grzes, Katarzyna M.; Ellis, Lucy; Epemolu, Ola; Shimamura, Satoko; Bantscheff, Marcus; Grandi, Paola; Read, Kevin D.; Cantrell, Doreen A.; Rocha, Sonia; Ciulli, Alessio (Lead / Corresponding author).

In: Nature Communications, Vol. 7, 13312, 04.11.2016, p. 1-12.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition

AU - Frost, Julianty

AU - Galdeano, Carles

AU - Soares, Pedro

AU - Gadd, Morgan S.

AU - Grzes, Katarzyna M.

AU - Ellis, Lucy

AU - Epemolu, Ola

AU - Shimamura, Satoko

AU - Bantscheff, Marcus

AU - Grandi, Paola

AU - Read, Kevin D.

AU - Cantrell, Doreen A.

AU - Rocha, Sonia

AU - Ciulli, Alessio

N1 - This work was supported by the European Research Council ERC-2012-StG-311460 DrugE3CRLs (Starting Grant to A.C.), the UK Biotechnology and Biological Sciences Research Council BBSRC BB/G023123/2 (David Phillips Fellowship to A.C.), the European Commission PIEF-GA-2012-328030 (Marie-Curie Intra-European Fellowship to C.G.), the Wellcome Trust (PhD Studentship 102398/Z/13/Z to J.F., and strategic awards 100476/Z/12/Z for biophysics and drug discovery and 094090/Z/10/Z for structural biology and X-ray crystallography to BCDD) and the Fundac¸a˜o para a Cieˆncia e Tecnologia FCT SFRH/BD/101598/2014 (PhD Studentship to P.S.). S.R. is funded by a Cancer Research UK Senior fellowship C99667/A12918 with support of a Wellcome Trust strategic award 097945/B/11/Z.

PY - 2016/11/4

Y1 - 2016/11/4

N2 - Chemical strategies to using small molecules to stimulate hypoxia inducible factors (HIFs) activity and trigger a hypoxic response under normoxic conditions, such as iron chelators and inhibitors of prolyl hydroxylase domain (PHD) enzymes, have broad-spectrum activities and off-target effects. Here we disclose VH298, a potent VHL inhibitor that stabilizes HIF-α and elicits a hypoxic response via a different mechanism, that is the blockade of the VHL:HIF-α protein-protein interaction downstream of HIF-α hydroxylation by PHD enzymes. We show that VH298 engages with high affinity and specificity with VHL as its only major cellular target, leading to selective on-target accumulation of hydroxylated HIF-α in a concentration- and time-dependent fashion in different cell lines, with subsequent upregulation of HIF-target genes at both mRNA and protein levels. VH298 represents a high-quality chemical probe of the HIF signaling cascade and an attractive starting point to the development of potential new therapeutics targeting hypoxia signaling.

AB - Chemical strategies to using small molecules to stimulate hypoxia inducible factors (HIFs) activity and trigger a hypoxic response under normoxic conditions, such as iron chelators and inhibitors of prolyl hydroxylase domain (PHD) enzymes, have broad-spectrum activities and off-target effects. Here we disclose VH298, a potent VHL inhibitor that stabilizes HIF-α and elicits a hypoxic response via a different mechanism, that is the blockade of the VHL:HIF-α protein-protein interaction downstream of HIF-α hydroxylation by PHD enzymes. We show that VH298 engages with high affinity and specificity with VHL as its only major cellular target, leading to selective on-target accumulation of hydroxylated HIF-α in a concentration- and time-dependent fashion in different cell lines, with subsequent upregulation of HIF-target genes at both mRNA and protein levels. VH298 represents a high-quality chemical probe of the HIF signaling cascade and an attractive starting point to the development of potential new therapeutics targeting hypoxia signaling.

U2 - 10.1038/ncomms13312

DO - 10.1038/ncomms13312

M3 - Article

C2 - 27811928

VL - 7

SP - 1

EP - 12

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

M1 - 13312

ER -