Ras/MAPK signalling intensity defines subclonal fitness in a mouse model of hepatocellular carcinoma

Abstract

Quantitative differences in signal transduction are to date an understudied feature of tumour heterogeneity. The MAPK Erk pathway, which is activated in a large proportion of human tumours, is a prototypic example of distinct cell fates being driven by signal intensity. We have used primary hepatocyte precursors transformed with different dosages of an oncogenic form of Ras to model subclonal variations in MAPK signalling. Orthotopic allografts of Ras-transformed cells in immunocompromised mice gave rise to fast-growing aggressive tumours, both at the primary location and in the peritoneal cavity. Fluorescent labelling of cells expressing different oncogene levels, and consequently varying levels of MAPK Erk activation, highlighted the selection processes operating at the two sites of tumour growth. Indeed, significantly higher Ras expression was observed in primary as compared to secondary, metastatic sites, despite the apparent evolutionary trade-off of increased apoptotic death in the liver that correlated with high Ras dosage. Analysis of the immune tumour microenvironment at the two locations suggests that fast peritoneal tumour growth in the immunocompromised setting is abrogated in immunocompetent animals due to efficient antigen presentation by peritoneal dendritic cells. Furthermore, our data indicate that, in contrast to the metastatic-like outgrowth, strong MAPK signalling is required in the primary liver tumours to resist elimination by NK cells. Overall, this study describes a quantitative aspect of tumour heterogeneity and points to a potential vulnerability of a subtype of hepatocellular carcinoma as a function of MAPK Erk signalling intensity.

Data availability

The RNA-sequencing data have been deposited in the Gene Expression Omnibus (GEO, NCBI) repository, and are accessible through GEO Series accession number GSE180580. Raw data from figures 1 to 5 were deposited on Mendeley data at doi: 10.17632/73nbvs8925.1.

The following data sets were generated

Article and author information

Author details

  1. Anthony Lozano

    Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  2. Francois-Régis Souche

    Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  3. Carine Chavey

    Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  4. Valérie Dardalhon

    Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  5. Christel Ramirez

    Division of Tumor Biology and Immunology, Oncode Institute, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  6. Serena Vegna

    Division of Tumor Biology and Immunology, Oncode Institute, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  7. Guillaume Desandre

    Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  8. Anaïs Riviere

    Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  9. Amal Zine El Aabidine

    Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  10. Philippe Fort

    Centre de Recherche en Biologie Cellulaire de Montpellier, French National Centre for Scientific Research, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5997-8722
  11. Leila Akkari

    Division of Tumor Biology and Immunology, Oncode Institute, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  12. Urszula Hibner

    Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France
    For correspondence
    ula.hibner@igmm.cnrs.fr
    Competing interests
    The authors declare that no competing interests exist.
  13. Damien Grégoire

    Institut de Génétique Moléculaire de Montpellier, University of Montpellier, CNRS, Montpellier, France
    For correspondence
    damien.gregoire@igmm.cnrs.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1105-8115

Funding

SIRIC Montpellier Cancer (Grant INCa_Inserm_DGOS_12553)

  • Urszula Hibner

Grant HTE-ITMO Cancer (HTE201610)

  • Urszula Hibner

Association Francaise pour l'Etude du Foie

  • Damien Grégoire

Dutch Cancer Society (KWF 12049/2018-2)

  • Leila Akkari

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

Reviewing Editor

  1. Roger J Davis, University of Massachusetts Medical School, United States

Ethics

Animal experimentation: All reported animal procedures were carried out in accordance with the rules of the FrenchInstitutional Animal Care and Use Committee and European Community Council(2010/63/EU). Animal studies were approved by institutional ethical committee (Comitéd'éthique en expérimentation animale Languedoc-Roussillon (#36)) and by the Ministère del'Enseignement Supérieur, de la Recherche et de l'Innovation (APAFIS#11196-2018090515538313v2).

Version history

  1. Preprint posted: August 13, 2021 (view preprint)
  2. Received: December 13, 2021
  3. Accepted: January 18, 2023
  4. Accepted Manuscript published: January 19, 2023 (version 1)
  5. Version of Record published: February 1, 2023 (version 2)

Copyright

© 2023, Lozano 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.

Metrics

  • 988
    Page views
  • 118
    Downloads
  • 2
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Anthony Lozano
  2. Francois-Régis Souche
  3. Carine Chavey
  4. Valérie Dardalhon
  5. Christel Ramirez
  6. Serena Vegna
  7. Guillaume Desandre
  8. Anaïs Riviere
  9. Amal Zine El Aabidine
  10. Philippe Fort
  11. Leila Akkari
  12. Urszula Hibner
  13. Damien Grégoire
(2023)
Ras/MAPK signalling intensity defines subclonal fitness in a mouse model of hepatocellular carcinoma
eLife 12:e76294.
https://doi.org/10.7554/eLife.76294

Share this article

https://doi.org/10.7554/eLife.76294

Further reading

    1. Cancer Biology
    2. Structural Biology and Molecular Biophysics
    Johannes Paladini, Annalena Maier ... Stephan Grzesiek
    Research Article

    Abelson tyrosine kinase (Abl) is regulated by the arrangement of its regulatory core, consisting sequentially of the SH3, SH2, and kinase (KD) domains, where an assembled or disassembled core corresponds to low or high kinase activity, respectively. It was recently established that binding of type II ATP site inhibitors, such as imatinib, generates a force from the KD N-lobe onto the SH3 domain and in consequence disassembles the core. Here, we demonstrate that the C-terminal αI-helix exerts an additional force toward the SH2 domain, which correlates both with kinase activity and type II inhibitor-induced disassembly. The αI-helix mutation E528K, which is responsible for the ABL1 malformation syndrome, strongly activates Abl by breaking a salt bridge with the KD C-lobe and thereby increasing the force onto the SH2 domain. In contrast, the allosteric inhibitor asciminib strongly reduces Abl’s activity by fixating the αI-helix and reducing the force onto the SH2 domain. These observations are explained by a simple mechanical model of Abl activation involving forces from the KD N-lobe and the αI-helix onto the KD/SH2SH3 interface.

    1. Biochemistry and Chemical Biology
    2. Cancer Biology
    Litong Nie, Chao Wang ... Junjie Chen
    Research Article

    Poly(ADP-ribose)ylation or PARylation by PAR polymerase 1 (PARP1) and dePARylation by poly(ADP-ribose) glycohydrolase (PARG) are equally important for the dynamic regulation of DNA damage response. PARG, the most active dePARylation enzyme, is recruited to sites of DNA damage via pADPr-dependent and PCNA-dependent mechanisms. Targeting dePARylation is considered an alternative strategy to overcome PARP inhibitor resistance. However, precisely how dePARylation functions in normal unperturbed cells remains elusive. To address this challenge, we conducted multiple CRISPR screens and revealed that dePARylation of S phase pADPr by PARG is essential for cell viability. Loss of dePARylation activity initially induced S-phase-specific pADPr signaling, which resulted from unligated Okazaki fragments and eventually led to uncontrolled pADPr accumulation and PARP1/2-dependent cytotoxicity. Moreover, we demonstrated that proteins involved in Okazaki fragment ligation and/or base excision repair regulate pADPr signaling and cell death induced by PARG inhibition. In addition, we determined that PARG expression is critical for cellular sensitivity to PARG inhibition. Additionally, we revealed that PARG is essential for cell survival by suppressing pADPr. Collectively, our data not only identify an essential role for PARG in normal proliferating cells but also provide a potential biomarker for the further development of PARG inhibitors in cancer therapy.