Article Text

Download PDFPDF
Original research
Escape from cell-cell and cell-matrix adhesion dependence underscores disease progression in gastric cancer organoid models
  1. Yin Tong1,2,
  2. Priscilla S W Cheng1,
  3. Chung Sze Or1,
  4. Sarah S K Yue1,
  5. Hoi Cheong Siu1,
  6. Siu Lun Ho1,
  7. Simon Y K Law3,
  8. Wai Yin Tsui1,
  9. Dessy Chan1,
  10. Stephanie Ma4,
  11. Siu Po Lee1,
  12. Annie S Y Chan1,
  13. April S Chan1,
  14. Shui Wa Yun1,
  15. Ho Sang Hui1,
  16. Siu Tsan Yuen1,5,
  17. Suet Yi Leung1,2,6,
  18. Helen H N Yan1,2
  1. 1Department of Pathology, School of Clinical Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam, Hong Kong SAR, China
  2. 2Centre for Oncology and Immunology, Hong Kong Science Park, Hong Kong SAR, China
  3. 3Department of Surgery, School of Clinical Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam, Hong Kong SAR, China
  4. 4School of Biomedical Sciences, The University of Hong Kong, Pokfulam, Hong Kong SAR, China
  5. 5Department of Pathology, St. Paul's Hospital, No. 2, Eastern Hosptial Road, Causeway Bay, Hong Kong SAR, China
  6. 6The Jockey Club Centre for Clinical Innovation and Discovery, LKS Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China
  1. Correspondence to Dr Suet Yi Leung, Department of Pathology, School of Clinical Medicine, The University of Hong Kong, Queen Mary Hospital, Pokfulam, Hong Kong SAR, China; suetyi{at}hku.hk; Dr Helen H N Yan; yanhelen{at}hku.hk

Abstract

Objective Cell-cell (CC) and cell-matrix (CM) adhesions are essential for epithelial cell survival, yet dissociation-induced apoptosis is frequently circumvented in malignant cells.

Design We explored CC and CM dependence in 58 gastric cancer (GC) organoids by withdrawing either ROCK inhibitor, matrix or both to evaluate their tumorigenic potential in terms of apoptosis resistance, correlation with oncogenic driver mutations and clinical behaviour. We performed mechanistic studies to determine the role of diffuse-type GC drivers: ARHGAP fusions, RHOA and CDH1, in modulating CC (CCi) or CM (CMi) adhesion independence.

Results 97% of the tumour organoids were CMi, 66% were CCi and 52% were resistant to double withdrawal (CCi/CMi), while normal organoids were neither CMi nor CCi. Clinically, the CCi/CMi phenotype was associated with an infiltrative tumour edge and advanced tumour stage. Moreover, the CCi/CMi transcriptome signature was associated with poor patient survival when applied to three public GC datasets. CCi/CMi and CCi phenotypes were enriched in diffuse-type GC organoids, especially in those with oncogenic driver perturbation of RHO signalling via RHOA mutation or ARHGAP fusions. Inducible knockout of ARHGAP fusions in CCi/CMi tumour organoids led to resensitisation to CC/CM dissociation-induced apoptosis, upregulation of focal adhesion and tight junction genes, partial reversion to a more normal cystic phenotype and inhibited xenograft formation. Normal gastric organoids engineered with CDH1 or RHOA mutations became CMi or CCi, respectively.

Conclusions The CCi/CMi phenotype has a critical role in malignant transformation and tumour progression, offering new mechanistic information on RHO-ROCK pathway inhibition that contributes to GC pathogenicity.

  • GASTRIC CANCER
  • CELL ADHESION
  • CELL MATRIX INTERACTION
  • APOPTOSIS

Data availability statement

Data are available in a public, open access repository. The RNA sequencing data have been deposited into the European Genome-Phenome Archive with accession number EGAS00001006252 (https://www.ebi.ac.uk/ega/studies/EGAS00001006252). Application to a data access committee as a standard procedure in the ega for data access containing human genetic data is needed, for the use of data for medical research by bona fide researchers.

Statistics from Altmetric.com

Data availability statement

Data are available in a public, open access repository. The RNA sequencing data have been deposited into the European Genome-Phenome Archive with accession number EGAS00001006252 (https://www.ebi.ac.uk/ega/studies/EGAS00001006252). Application to a data access committee as a standard procedure in the ega for data access containing human genetic data is needed, for the use of data for medical research by bona fide researchers.

View Full Text

Footnotes

  • YT, PSWC and CSO contributed equally.

  • Correction notice This article has been corrected since it published Online First. The supplementary methods file has been replaced.

  • Contributors HHNY and SYL conceived, designed and supervised the study. YT, PSWC, CSO, SSKY, HHNY, SLH, WYT, DC, SPL, ASYC, SWY, HSH, STY and SYL collected data, performed experiments and/or analysed data. PSWC performed the ARHGAP fusion KO studies. CSO performed the CC and CM dependence assay in tumour organoids. YT and HCS performed the bioinformatics data analysis. SL contributed clinical data and samples. SM assisted with animal experiments. HHNY, SYL and YT wrote the manuscript with assistance from PSWC, CSO and ASC. All authors contributed comments and approval for the manuscript. HHNY and SYL are the guarantors.

  • Funding This work was substantially supported by a theme-based research grant (Project No. T12-710/16-R) from the Research Grants Council of the Hong Kong SAR, China, and partly supported by the Centre for Oncology and Immunology under the Health@InnoHK Initiative funded by the Innovation and Technology Commission, The Government of Hong Kong SAR, China. Functional characterisation of gastric organoids was partially supported by a General Research Fund (Project No. 17108616) from the Research Grants Council of the Hong Kong SAR, China.

  • Competing interests SYL and STY have received research sponsorships from Pfizer, Merck, Servier. The other authors declare no competing interests.

  • Patient and public involvement Patients and/or the public were not involved in the design, or conduct, or reporting, or dissemination plans of this research.

  • Provenance and peer review Not commissioned; externally peer reviewed.

  • Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.

Request Permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.