USP28 deletion and small molecule inhibition destabilises c-MYC and elicits regression of squamous cell lung carcinoma

  1. E Josue Ruiz
  2. Adan Pinto-Fernandez
  3. Andrew P Turnbull
  4. Linxiang Lan
  5. Thomas M Charlton
  6. Hannah C Scott
  7. Andreas Damianou
  8. George Vere
  9. Eva M Riising
  10. Clive Da Costa
  11. Wojciech W Krajewski
  12. David Guerin
  13. Jeffrey D Kearns
  14. Stephanos Ioannidis
  15. Marie Katz
  16. Crystal McKinnon
  17. Jonathan O'Connell
  18. Natalia Moncaut
  19. Ian Rosewell
  20. Emma Nye
  21. Neil Jones
  22. Claire Heride
  23. Malte Gersch
  24. Min Wu
  25. Christopher J Dinsmore
  26. Tim R Hammonds
  27. Sunkyu Kim
  28. David Komander
  29. Sylvie Urbe
  30. Michael J Clague
  31. Benedikt M Kessler  Is a corresponding author
  32. Axel Behrens  Is a corresponding author
  1. The Francis Crick Institute, United Kingdom
  2. University of Oxford, United Kingdom
  3. CRUK Therapeutic Discovery Laboratories, United Kingdom
  4. Forma Therapeutics, United States
  5. Novartis Institutes of BioMedical Research, United States
  6. FORMA Therapeutics, United States
  7. University College London, United Kingdom
  8. Francis Crick Institute, United Kingdom
  9. Max Planck Institut, Dortmund, Germany
  10. Walter and Eliza Hall Institute of Medical Research, Australia
  11. Liverpool University, United Kingdom

Abstract

Lung squamous cell carcinoma (LSCC) is a considerable global health burden, with an incidence of over 600,000 cases per year. Treatment options are limited, and patient 5-year survival rate is less than 5%. The ubiquitin specific protease 28 (USP28) has been implicated in tumorigenesis through its stabilization of the oncoproteins c-MYC, c-JUN and Dp63. Here, we show that genetic inactivation of Usp28 induced regression of established murine LSCC lung tumours. We developed a small molecule that inhibits USP28 activity in the low nanomole range. While displaying cross-reactivity against the closest homologue USP25, this inhibitor showed a high degree of selectivity over other deubiquitinases. USP28 inhibitor treatment resulted in a dramatic decrease in c-MYC, c-JUN and Dp63 proteins levels and consequently induced substantial regression of autochthonous murine LSCC tumors and human LSCC xenografts, thereby phenocopying the effect observed by genetic deletion. Thus, USP28 may represent a promising therapeutic target for the treatment of squamous cell lung carcinoma.

Data availability

For all figures with graphs we provide source data files in the Supplemental Information section. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with an internal ID of PXD469830.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. E Josue Ruiz

    King's College London, The Francis Crick Institute, London, United Kingdom
    Competing interests
    No competing interests declared.
  2. Adan Pinto-Fernandez

    Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  3. Andrew P Turnbull

    London Bioscience Innovation Centre, CRUK Therapeutic Discovery Laboratories, London, United Kingdom
    Competing interests
    Andrew P Turnbull, Andrew P Turnbull is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared.APT declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  4. Linxiang Lan

    King's College London, The Francis Crick Institute, London, United Kingdom
    Competing interests
    No competing interests declared.
  5. Thomas M Charlton

    Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  6. Hannah C Scott

    Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  7. Andreas Damianou

    Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
  8. George Vere

    Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1154-5212
  9. Eva M Riising

    King's College London, The Francis Crick Institute, London, United Kingdom
    Competing interests
    No competing interests declared.
  10. Clive Da Costa

    King's College London, The Francis Crick Institute, London, United Kingdom
    Competing interests
    No competing interests declared.
  11. Wojciech W Krajewski

    London Bioscience Innovation Centre, CRUK Therapeutic Discovery Laboratories, London, United Kingdom
    Competing interests
    Wojciech W Krajewski, Wojciech W Krajewski is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared. WWK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  12. David Guerin

    Watertown, Massachusetts 02472, USA, Forma Therapeutics, Boston, United States
    Competing interests
    David Guerin, Dave Guerin is affiliated with Constellation Pharmaceuticals (Boston, USA), for which no financial interests have been declared.DG declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  13. Jeffrey D Kearns

    Translational Medicine, PK Sciences Modeling & Simulation, Novartis Institutes of BioMedical Research, Cambridge, United States
    Competing interests
    Jeffrey D Kearns, Jeffrey Kearns is affiliated with the Novartis Institutes for BioMedical Research (Boston, USA), for which no financial interests have been declared.JK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7261-5096
  14. Stephanos Ioannidis

    Forma Therapeutics, Watertown, MA, United States
    Competing interests
    Stephanos Ioannidis, Stephanos Ioannidis is affiliated with H3 Biomedicine (Cambridge, MA, USA), for which no financial interests have been declared.SI declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  15. Marie Katz

    FORMA Therapeutics, Boston, United States
    Competing interests
    Marie Katz, Marie Katz is affiliated with Valo Health (Boston, USA), for which no financial interests have been declared.MK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  16. Crystal McKinnon

    FORMA Therapeutics, Boston, United States
    Competing interests
    Crystal McKinnon, Crystal McKinnon is affiliated with Valo Health (Boston, USA), for which no financial interests have been declared.CM declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  17. Jonathan O'Connell

    Forma Therapeutics, Watertown, MA, United States
    Competing interests
    Jonathan O'Connell, Johnathan O'Connell is affiliated with Valo Health (Boston, USA), for which no financial interests have been declared.JOC declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  18. Natalia Moncaut

    University College London, London, United Kingdom
    Competing interests
    No competing interests declared.
  19. Ian Rosewell

    Genetic Modification Service, The Francis Crick Institute, Londonj, United Kingdom
    Competing interests
    No competing interests declared.
  20. Emma Nye

    Experimental Histopathology Laboratory, Francis Crick Institute, London, United Kingdom
    Competing interests
    No competing interests declared.
  21. Neil Jones

    London Bioscience Innovation Centre, CRUK Therapeutic Discovery Laboratories, London, United Kingdom
    Competing interests
    Neil Jones, Neil Jones is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared.NJ declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  22. Claire Heride

    Target Validation and Disease Positioning, CRUK Therapeutic Discovery Laboratories, London, United Kingdom
    Competing interests
    Claire Heride, Claire Heride is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared.CH declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1927-9577
  23. Malte Gersch

    Max Planck Institut, Dortmund, Dortmund, Germany
    Competing interests
    No competing interests declared.
  24. Min Wu

    FORMA Therapeutics, Boston, United States
    Competing interests
    Min Wu, Min Wu is affiliated with Disc Medicine (Cambridge, MA, USA), for which no financial interests have been declared.MW declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  25. Christopher J Dinsmore

    FORMA Therapeutics, Boston, United States
    Competing interests
    Christopher J Dinsmore, Christopher J Dinsmore is affiliated with Disc Medicine (Cambridge, MA, USA), for which no financial interests have been declared.CJD declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  26. Tim R Hammonds

    Target Validation and Disease Positioning, CRUK Therapeutic Discovery Laboratories, London, United Kingdom
    Competing interests
    Tim R Hammonds, Tim R Hammonds is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared.TRH declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  27. Sunkyu Kim

    FORMA Therapeutics, Boston, United States
    Competing interests
    Sunkyu Kim, Sunkyu Kim is affiliated with Incyte (Wilmington, DE, USA), for which no financial interests have been declared.SK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  28. David Komander

    Walter and Eliza Hall Institute of Medical Research, Parkville, Australia
    Competing interests
    David Komander, DK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  29. Sylvie Urbe

    Liverpool University, Liverpool, United Kingdom
    Competing interests
    Sylvie Urbe, SU declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
  30. Michael J Clague

    Liverpool University, Liverpool, United Kingdom
    Competing interests
    Michael J Clague, MJC declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3355-9479
  31. Benedikt M Kessler

    Target Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
    For correspondence
    benedikt.kessler@ndm.ox.ac.uk
    Competing interests
    Benedikt M Kessler, BMK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8160-2446
  32. Axel Behrens

    King's College London, The Francis Crick Institute, London, United Kingdom
    For correspondence
    axel.behrens@icr.ac.uk
    Competing interests
    Axel Behrens, AB declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1557-1143

Funding

Forma Therapeutics (2013-2019)

  • E Josue Ruiz
  • Adan Pinto-Fernandez
  • Andrew P Turnbull
  • Linxiang Lan
  • Thomas M Charlton
  • Hannah C Scott
  • Andreas Damianou
  • George Vere
  • Eva M Riising
  • Clive Da Costa
  • Wojciech W Krajewski
  • David Guerin
  • Jeffrey D Kearns
  • Stephanos Ioannidis
  • Marie Katz
  • Crystal McKinnon
  • Jonathan O'Connell
  • Natalia Moncaut
  • Ian Rosewell
  • Emma Nye
  • Neil Jones
  • Claire Heride
  • Malte Gersch
  • Min Wu
  • Christopher J Dinsmore
  • Tim R Hammonds
  • Sunkyu Kim
  • David Komander
  • Sylvie Urbe
  • Michael J Clague
  • Benedikt M Kessler
  • Axel Behrens

Cancer Research UK Manchester Centre (FC001039)

  • Axel Behrens

Medical Research Council (FC001039)

  • Axel Behrens

Wellcome Trust (FC001039)

  • Axel Behrens

Wellcome Trust (097813/Z/11/Z)

  • Benedikt M Kessler

Engineering and Physical Sciences Research Council (EP/N034295/1)

  • Benedikt M Kessler

John Fell Fund, University of Oxford (133/075)

  • Benedikt M Kessler

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal experiments were approved by the Francis Crick Institute Animal Ethics Committee and conformed to UK Home Office regulations under the Animals (Scientific Procedures) Act 1986 including Amendment Regulations 2012.

Reviewing Editor

  1. Erica A Golemis, Fox Chase Cancer Center, United States

Publication history

  1. Received: June 24, 2021
  2. Accepted: October 10, 2021
  3. Accepted Manuscript published: October 12, 2021 (version 1)
  4. Version of Record published: October 28, 2021 (version 2)

Copyright

© 2021, Ruiz et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. E Josue Ruiz
  2. Adan Pinto-Fernandez
  3. Andrew P Turnbull
  4. Linxiang Lan
  5. Thomas M Charlton
  6. Hannah C Scott
  7. Andreas Damianou
  8. George Vere
  9. Eva M Riising
  10. Clive Da Costa
  11. Wojciech W Krajewski
  12. David Guerin
  13. Jeffrey D Kearns
  14. Stephanos Ioannidis
  15. Marie Katz
  16. Crystal McKinnon
  17. Jonathan O'Connell
  18. Natalia Moncaut
  19. Ian Rosewell
  20. Emma Nye
  21. Neil Jones
  22. Claire Heride
  23. Malte Gersch
  24. Min Wu
  25. Christopher J Dinsmore
  26. Tim R Hammonds
  27. Sunkyu Kim
  28. David Komander
  29. Sylvie Urbe
  30. Michael J Clague
  31. Benedikt M Kessler
  32. Axel Behrens
(2021)
USP28 deletion and small molecule inhibition destabilises c-MYC and elicits regression of squamous cell lung carcinoma
eLife 10:e71596.
https://doi.org/10.7554/eLife.71596

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