Inhibitors of Rho kinases (ROCK) induce multiple mitotic defects and synthetic lethality in BRCA2-deficient cells

  1. Julieta Martino
  2. Sebastián Omar Siri
  3. Nicolás Luis Calzetta
  4. Natalia Soledad Paviolo
  5. Cintia Garro
  6. Maria F Pansa
  7. Sofía Carbajosa
  8. Aaron C Brown
  9. José Luis Bocco
  10. Israel Gloger
  11. Gerard Drewes
  12. Kevin P Madauss
  13. Gastón Soria  Is a corresponding author
  14. Vanesa Gottifredi  Is a corresponding author
  1. Fundación Instituto Leloir-CONICET, Argentina
  2. Universidad Nacional de Córdoba, Argentina
  3. Maine Medical Center Research Institute, United States
  4. GlaxoSmithKline, United States
  5. GlaxoSmithKline, United Kingdom

Abstract

The trapping of Poly-ADP-ribose polymerase (PARP) on DNA caused by PARP inhibitors (PARPi) triggers acute DNA replication stress and synthetic lethality (SL) in BRCA2-deficient cells. Hence, DNA damage is accepted as a prerequisite for SL in BRCA2-deficient cells. In contrast, here we show that inhibiting ROCK in BRCA2-deficient cells triggers SL independently from acute replication stress. Such SL is preceded by polyploidy and binucleation resulting from cytokinesis failure. Such initial mitosis abnormalities are followed by other M-phase defects, including anaphase bridges and abnormal mitotic figures associated with multipolar spindles, supernumerary centrosomes and multinucleation. SL was also triggered by inhibiting Citron Rho-interacting kinase, another enzyme that, similarly to ROCK, regulates cytokinesis. Together, these observations demonstrate that cytokinesis failure triggers mitotic abnormalities and SL in BRCA2-deficient cells. Furthermore, the prevention of mitotic entry by depletion of Early mitotic inhibitor 1 (EMI1) augmented the survival of BRCA2-deficient cells treated with ROCK inhibitors, thus reinforcing the association between M-phase and cell death in BRCA2-deficient cells. This novel SL differs from the one triggered by PARPi and uncovers mitosis as an Achilles heel of BRCA2-deficient cells.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file

The following data sets were generated

Article and author information

Author details

  1. Julieta Martino

    Fundación Instituto Leloir-CONICET, Buenos Aires, Argentina
    Competing interests
    No competing interests declared.
  2. Sebastián Omar Siri

    Fundación Instituto Leloir-CONICET, Buenos Aires, Argentina
    Competing interests
    No competing interests declared.
  3. Nicolás Luis Calzetta

    Fundación Instituto Leloir-CONICET, Buenos Aires, Argentina
    Competing interests
    No competing interests declared.
  4. Natalia Soledad Paviolo

    Fundación Instituto Leloir-CONICET, Buenos Aires, Argentina
    Competing interests
    No competing interests declared.
  5. Cintia Garro

    Departamento de Bioquímica Clínica, Universidad Nacional de Córdoba, Córdoba, Argentina
    Competing interests
    No competing interests declared.
  6. Maria F Pansa

    Departamento de Bioquímica Clínica, Universidad Nacional de Córdoba, Córdoba, Argentina
    Competing interests
    Maria F Pansa, is affiliated with GlaxoSmithKline and has no other competing interests to declare..
  7. Sofía Carbajosa

    Departamento de Bioquímica Clínica, Universidad Nacional de Córdoba, Córdoba, Argentina
    Competing interests
    No competing interests declared.
  8. Aaron C Brown

    Center for Molecular Medicine, Maine Medical Center Research Institute, Scarborough, United States
    Competing interests
    No competing interests declared.
  9. José Luis Bocco

    Departamento de Bioquímica Clínica, Universidad Nacional de Córdoba, Córdoba, Argentina
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9682-1270
  10. Israel Gloger

    GlaxoSmithKline, East Durham, United States
    Competing interests
    Israel Gloger, is affiliated with GlaxoSmithKline and has no other competing interests to declare..
  11. Gerard Drewes

    Global Health R and D, GlaxoSmithKline, Stevenage, United Kingdom
    Competing interests
    Gerard Drewes, is affiliated with GlaxoSmithKline and has no other competing interests to declare..
  12. Kevin P Madauss

    Global Health R and D, GlaxoSmithKline, Stevenage, United Kingdom
    Competing interests
    Kevin P Madauss, is affiliated with GlaxoSmithKline and has no other competing interests to declare..
  13. Gastón Soria

    Departamento de Bioquímica Clínica, Universidad Nacional de Córdoba, Córdoba, Argentina
    For correspondence
    gsoria29@gmail.com
    Competing interests
    No competing interests declared.
  14. Vanesa Gottifredi

    Fundación Instituto Leloir-CONICET, Buenos Aires, Argentina
    For correspondence
    vgottifredi@leloir.org.ar
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9656-5951

Funding

GlaxoSmithKline (PAE-GLAXO 2014-0005)

  • José Luis Bocco

GlaxoSmithKline (PCE-GSK 2017-0032)

  • Gastón Soria

Fondo para la Investigación Científica y Tecnológica

  • José Luis Bocco
  • Gastón Soria

Agencia Nacional de Promoción Científica y Tecnológica (PICT 2018-01857)

  • Vanesa Gottifredi

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

Reviewing Editor

  1. Robert W. Sobol, Brown University, United States

Version history

  1. Received: May 13, 2022
  2. Preprint posted: June 28, 2022 (view preprint)
  3. Accepted: April 18, 2023
  4. Accepted Manuscript published: April 19, 2023 (version 1)
  5. Version of Record published: May 15, 2023 (version 2)
  6. Version of Record updated: November 14, 2023 (version 3)

Copyright

© 2023, Martino 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. Julieta Martino
  2. Sebastián Omar Siri
  3. Nicolás Luis Calzetta
  4. Natalia Soledad Paviolo
  5. Cintia Garro
  6. Maria F Pansa
  7. Sofía Carbajosa
  8. Aaron C Brown
  9. José Luis Bocco
  10. Israel Gloger
  11. Gerard Drewes
  12. Kevin P Madauss
  13. Gastón Soria
  14. Vanesa Gottifredi
(2023)
Inhibitors of Rho kinases (ROCK) induce multiple mitotic defects and synthetic lethality in BRCA2-deficient cells
eLife 12:e80254.
https://doi.org/10.7554/eLife.80254

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https://doi.org/10.7554/eLife.80254

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