Thermal proteome profiling of breast cancer cells reveals proteasomal activation by CDK4/6 inhibitor palbociclib

Teemu P. Miettinen, Julien Peltier, Anetta Härtlova, Marek Gierliński, Valerie M. Jansen, Matthias Trost (Lead / Corresponding author), Mikael Björklund (Lead / Corresponding author)

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Abstract

Palbociclib is a CDK4/6 inhibitor approved for metastatic estrogen receptor-positive breast cancer. In addition to G1 cell cycle arrest, palbociclib treatment results in cell senescence, a phenotype that is not readily explained by CDK4/6 inhibition. In order to identify a molecular mechanism responsible for palbociclib-induced senescence, we performed thermal proteome profiling of MCF7 breast cancer cells. In addition to affecting known CDK4/6 targets, palbociclib induces a thermal stabilization of the 20S proteasome, despite not directly binding to it. We further show that palbociclib treatment increases proteasome activity independently of the ubiquitin pathway. This leads to cellular senescence, which can be counteracted by proteasome inhibitors. Palbociclib-induced proteasome activation and senescence is mediated by reduced proteasomal association of ECM29. Loss of ECM29 activates the proteasome, blocks cell proliferation, and induces a senescence-like phenotype. Finally, we find that ECM29 mRNA levels are predictive of relapse-free survival in breast cancer patients treated with endocrine therapy. In conclusion, thermal proteome profiling identifies the proteasome and ECM29 protein as mediators of palbociclib activity in breast cancer cells.

Original languageEnglish
Article numbere98359
Pages (from-to)1-19
Number of pages19
JournalEMBO Journal
Volume37
Issue number10
Early online date18 Apr 2018
DOIs
Publication statusPublished - 15 May 2018

Fingerprint

Proteome
Hot Temperature
Proteasome Endopeptidase Complex
Chemical activation
Cells
Breast Neoplasms
Cell Aging
G1 Phase Cell Cycle Checkpoints
Phenotype
Proteasome Inhibitors
Cell proliferation
Ubiquitin
palbociclib
Estrogen Receptors
Therapeutics
Stabilization
Cell Proliferation
Association reactions
Recurrence
Messenger RNA

Keywords

  • breast cancer
  • CDK4
  • palbociclib
  • proteasome
  • thermal proteome profiling

Cite this

Miettinen, T. P., Peltier, J., Härtlova, A., Gierliński, M., Jansen, V. M., Trost, M., & Björklund, M. (2018). Thermal proteome profiling of breast cancer cells reveals proteasomal activation by CDK4/6 inhibitor palbociclib. EMBO Journal, 37(10), 1-19. [e98359]. https://doi.org/10.15252/embj.201798359
Miettinen, Teemu P. ; Peltier, Julien ; Härtlova, Anetta ; Gierliński, Marek ; Jansen, Valerie M. ; Trost, Matthias ; Björklund, Mikael. / Thermal proteome profiling of breast cancer cells reveals proteasomal activation by CDK4/6 inhibitor palbociclib. In: EMBO Journal. 2018 ; Vol. 37, No. 10. pp. 1-19.
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abstract = "Palbociclib is a CDK4/6 inhibitor approved for metastatic estrogen receptor-positive breast cancer. In addition to G1 cell cycle arrest, palbociclib treatment results in cell senescence, a phenotype that is not readily explained by CDK4/6 inhibition. In order to identify a molecular mechanism responsible for palbociclib-induced senescence, we performed thermal proteome profiling of MCF7 breast cancer cells. In addition to affecting known CDK4/6 targets, palbociclib induces a thermal stabilization of the 20S proteasome, despite not directly binding to it. We further show that palbociclib treatment increases proteasome activity independently of the ubiquitin pathway. This leads to cellular senescence, which can be counteracted by proteasome inhibitors. Palbociclib-induced proteasome activation and senescence is mediated by reduced proteasomal association of ECM29. Loss of ECM29 activates the proteasome, blocks cell proliferation, and induces a senescence-like phenotype. Finally, we find that ECM29 mRNA levels are predictive of relapse-free survival in breast cancer patients treated with endocrine therapy. In conclusion, thermal proteome profiling identifies the proteasome and ECM29 protein as mediators of palbociclib activity in breast cancer cells.",
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Miettinen, TP, Peltier, J, Härtlova, A, Gierliński, M, Jansen, VM, Trost, M & Björklund, M 2018, 'Thermal proteome profiling of breast cancer cells reveals proteasomal activation by CDK4/6 inhibitor palbociclib', EMBO Journal, vol. 37, no. 10, e98359, pp. 1-19. https://doi.org/10.15252/embj.201798359

Thermal proteome profiling of breast cancer cells reveals proteasomal activation by CDK4/6 inhibitor palbociclib. / Miettinen, Teemu P.; Peltier, Julien; Härtlova, Anetta; Gierliński, Marek; Jansen, Valerie M.; Trost, Matthias (Lead / Corresponding author); Björklund, Mikael (Lead / Corresponding author).

In: EMBO Journal, Vol. 37, No. 10, e98359, 15.05.2018, p. 1-19.

Research output: Contribution to journalArticle

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AU - Miettinen, Teemu P.

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AU - Härtlova, Anetta

AU - Gierliński, Marek

AU - Jansen, Valerie M.

AU - Trost, Matthias

AU - Björklund, Mikael

N1 - This work was funded by Medical Research Council UK (MC_UU_12016/5 to M.T.), the BBSRC (BB/L008874/1), and the pharmaceutical companies supporting the Division of Signal Transduction Therapy (DSTT) (AstraZeneca, Boehringer-Ingelheim, GlaxoSmithKline, Janssen Pharmaceuticals, Merck KGaA, and Pfizer). M.T. thanks Thermo-Fisher Scientific for support with the TMT Research Award. The School of Life Sciences Data Analysis Group is funded by the Wellcome Trust grant 097945/Z/11/Z. T.P.M. is supported by the Wellcome Trust Sir Henry Postdoctoral Fellowship (grant number 110275/Z/15/Z). V.M.J is supported by the Conquer Cancer Foundation of ASCO Young Investigator Award (8364), Komen Post-Doctoral Award (15329319), and the Vanderbilt Clinical Oncology Research Career Development Program (2K12CA090625-17).

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N2 - Palbociclib is a CDK4/6 inhibitor approved for metastatic estrogen receptor-positive breast cancer. In addition to G1 cell cycle arrest, palbociclib treatment results in cell senescence, a phenotype that is not readily explained by CDK4/6 inhibition. In order to identify a molecular mechanism responsible for palbociclib-induced senescence, we performed thermal proteome profiling of MCF7 breast cancer cells. In addition to affecting known CDK4/6 targets, palbociclib induces a thermal stabilization of the 20S proteasome, despite not directly binding to it. We further show that palbociclib treatment increases proteasome activity independently of the ubiquitin pathway. This leads to cellular senescence, which can be counteracted by proteasome inhibitors. Palbociclib-induced proteasome activation and senescence is mediated by reduced proteasomal association of ECM29. Loss of ECM29 activates the proteasome, blocks cell proliferation, and induces a senescence-like phenotype. Finally, we find that ECM29 mRNA levels are predictive of relapse-free survival in breast cancer patients treated with endocrine therapy. In conclusion, thermal proteome profiling identifies the proteasome and ECM29 protein as mediators of palbociclib activity in breast cancer cells.

AB - Palbociclib is a CDK4/6 inhibitor approved for metastatic estrogen receptor-positive breast cancer. In addition to G1 cell cycle arrest, palbociclib treatment results in cell senescence, a phenotype that is not readily explained by CDK4/6 inhibition. In order to identify a molecular mechanism responsible for palbociclib-induced senescence, we performed thermal proteome profiling of MCF7 breast cancer cells. In addition to affecting known CDK4/6 targets, palbociclib induces a thermal stabilization of the 20S proteasome, despite not directly binding to it. We further show that palbociclib treatment increases proteasome activity independently of the ubiquitin pathway. This leads to cellular senescence, which can be counteracted by proteasome inhibitors. Palbociclib-induced proteasome activation and senescence is mediated by reduced proteasomal association of ECM29. Loss of ECM29 activates the proteasome, blocks cell proliferation, and induces a senescence-like phenotype. Finally, we find that ECM29 mRNA levels are predictive of relapse-free survival in breast cancer patients treated with endocrine therapy. In conclusion, thermal proteome profiling identifies the proteasome and ECM29 protein as mediators of palbociclib activity in breast cancer cells.

KW - breast cancer

KW - CDK4

KW - palbociclib

KW - proteasome

KW - thermal proteome profiling

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DO - 10.15252/embj.201798359

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JO - EMBO Journal

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