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

doi:10.1038/ncomms13312

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 language	English
Article number	13312
Pages (from-to)	1-12
Number of pages	12
Journal	Nature Communications
Volume	7
DOIs	https://doi.org/10.1038/ncomms13312
Publication status	Published - 4 Nov 2016

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Final published version, 1.67 MBLicence: CC BY

DrugE3CRL's: Probing Druggability of Multisubunit Complexes: E3 Cullin RING Ligases (ERC Starting Grant)
Ciulli, A. (Investigator)
COMMISSION OF THE EUROPEAN COMMUNITIES
1/05/13 → 30/04/18
Project: Research
Dissecting and Exploiting Molecular Recognition at Protein-Protein Interfaces (David Phillips Fellowship)
Ciulli, A. (Investigator)
Biotechnology and Biological Sciences Research Council
8/04/13 → 7/07/15
Project: Research
Design of Orthogonal Molecular Probes Targeting Engineered Von-Hippel Lindau (VHL) E3 Ubiquitin Ligase for the Control of Intercellular Protein Levels
Ciulli, A. (Investigator)
COMMISSION OF THE EUROPEAN COMMUNITIES
8/04/13 → 7/04/15
Project: Research

Small-molecule approaches to interrogate the druggability of the VHL E3 Cullin RING Ubiquitin Ligase
Soares, P. (Author), Ciulli, A. (Supervisor), 2018
Student thesis: Doctoral Thesis › Doctor of Philosophy

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Ciulli, Alessio
- Centre for Targeted Protein Degradation - Professor of Chemical and Structural Biology
Person: Academic

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Frost, J., Galdeano, C., Soares, P., Gadd, M. S., Grzes, K. M., Ellis, L., Epemolu, O., Shimamura, S., Bantscheff, M., Grandi, P., Read, K. D., Cantrell, D. A., Rocha, S., & Ciulli, A. (2016). Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition. Nature Communications, 7, 1-12. Article 13312. https://doi.org/10.1038/ncomms13312

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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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doi = "10.1038/ncomms13312",

language = "English",

volume = "7",

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journal = "Nature Communications",

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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

SN - 2041-1723

VL - 7

SP - 1

EP - 12

JO - Nature Communications

JF - Nature Communications

M1 - 13312

ER -

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

Abstract

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Projects

DrugE3CRL's: Probing Druggability of Multisubunit Complexes: E3 Cullin RING Ligases (ERC Starting Grant)

Dissecting and Exploiting Molecular Recognition at Protein-Protein Interfaces (David Phillips Fellowship)

Design of Orthogonal Molecular Probes Targeting Engineered Von-Hippel Lindau (VHL) E3 Ubiquitin Ligase for the Control of Intercellular Protein Levels

Student theses

Small-molecule approaches to interrogate the druggability of the VHL E3 Cullin RING Ubiquitin Ligase

Profiles

Ciulli, Alessio

Cite this