Re-expression of SMARCA4/BRG1 in Small Cell Carcinoma of Ovary, Hypercalcemic Type (SCCOHT) promotes an epithelial-like gene signature through an AP-1-dependent mechanism

  1. Krystal Ann Orlando
  2. Amber K Douglas
  3. Aierken Abudu
  4. Yemin Wang
  5. Basile Tessier-Cloutier
  6. Weiping Su
  7. Alec Peters
  8. Larry S Sherman
  9. Rayvon Moore
  10. Vinh Nguyen
  11. Gian Luca Negri
  12. Shane Colborne
  13. Gregg B Morin
  14. Friedrich Kommoss
  15. Jessica D Lang
  16. William PD Hendricks
  17. Elizabeth A Raupach
  18. Patrick Pirrotte
  19. David G Huntsman
  20. Jeffrey M Trent
  21. Joel S Parker
  22. Jesse R Raab  Is a corresponding author
  23. Bernard E Weissman  Is a corresponding author
  1. University of North Carolina at Chapel Hill, United States
  2. Michigan State University, United States
  3. University of British Columnia, Canada
  4. Oregon Health & Science University, United States
  5. British Columbia Cancer Agency, Canada
  6. Synlab MVZ Pathologie, Germany
  7. Translational Genomics Research Institute, United States
  8. British Columbia Cancer Research Institute, Canada

Abstract

Small cell carcinoma of the ovary, hypercalcemic type (SCCOHT) is a rare and aggressive form of ovarian cancer. SCCOHT tumors have inactivating mutations in SMARCA4 (BRG1), one of the two mutually exclusive ATPases of the SWI/SNF chromatin remodeling complex. To address the role that BRG1 loss plays in SCCOHT tumorigenesis, we performed integrative multi-omic analyses in SCCOHT cell lines +/- BRG1 re-expression. BRG1 re-expression induced a gene and protein signature similar to an epithelial cell and gained chromatin accessibility sites correlated with other epithelial originating TCGA tumors. Gained chromatin accessibility and BRG1 recruited sites were strongly enriched for transcription factor binding motifs of AP-1 family members. Furthermore, AP-1 motifs were enriched at the promoters of highly upregulated epithelial genes. Using a dominant negative AP-1 cell line, we found that both AP-1 DNA binding activity and BRG1 re-expression are necessary for the gene and protein expression of epithelial genes. Our study demonstrates that BRG1 re-expression drives an epithelial-like gene and protein signature in SCCOHT cells that depends upon by AP-1 activity.

Data availability

Raw fastq files and processed data have been deposited in Gene Expression Omnibus (GEO) database with the accession number: GSE151026. Proteomics data was deposited in PRIDE database (accession #PXD014134).

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

Article and author information

Author details

  1. Krystal Ann Orlando

    Department of Pathology and Laboratory Medicine; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Amber K Douglas

    Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Aierken Abudu

    Department of Mirobiology and Molecular Genetics, Michigan State University, East Lansing, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Yemin Wang

    Department of Pathology and Laboratory Medicine, University of British Columnia, Vancouver, Canada
    Competing interests
    The authors declare that no competing interests exist.
  5. Basile Tessier-Cloutier

    Department of Pathology and Laboratory Medicine, University of British Columnia, Vancouver, Canada
    Competing interests
    The authors declare that no competing interests exist.
  6. Weiping Su

    Cell, Developmental, and Cancer Biology, Oregon Health & Science University, Beaverton, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Alec Peters

    Cell, Developmental, and Cancer Biology, Oregon Health & Science University, Beaverton, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Larry S Sherman

    Division of Neuroscience, Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Rayvon Moore

    Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Vinh Nguyen

    Curriculum in Toxicology and Environmental Medicine; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Gian Luca Negri

    Michael Smith Genome Science Centre, British Columbia Cancer Agency, Vancouver, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7722-8888
  12. Shane Colborne

    Michael Smith Genome Science Centre, British Columbia Cancer Agency, Vancouver, Canada
    Competing interests
    The authors declare that no competing interests exist.
  13. Gregg B Morin

    Genome Sciences, British Columbia Cancer Agency, Vancouver, Canada
    Competing interests
    The authors declare that no competing interests exist.
  14. Friedrich Kommoss

    Institute of Pathology, Synlab MVZ Pathologie, Mannheim, Germany
    Competing interests
    The authors declare that no competing interests exist.
  15. Jessica D Lang

    Division of Integrated Cancer Genomics, Translational Genomics Research Institute, Phoenix, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9700-4785
  16. William PD Hendricks

    Division of Integrated Cancer Genomics, Translational Genomics Research Institute, Phoenix, United States
    Competing interests
    The authors declare that no competing interests exist.
  17. Elizabeth A Raupach

    Division of Integrated Cancer Genomics, Translational Genomics Research Institute, Phoenix, United States
    Competing interests
    The authors declare that no competing interests exist.
  18. Patrick Pirrotte

    Collaborative Center for Translational Mass Spectrometry, Translational Genomics Research Institute, Phoenix, United States
    Competing interests
    The authors declare that no competing interests exist.
  19. David G Huntsman

    British Columbia Cancer Research Institute, Vancouver, Canada
    Competing interests
    The authors declare that no competing interests exist.
  20. Jeffrey M Trent

    Division of Integrated Cancer Genomics, Translational Genomics Research Institute, Phoenix, United States
    Competing interests
    The authors declare that no competing interests exist.
  21. Joel S Parker

    Department of Genetics; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  22. Jesse R Raab

    Department of Genetics, University of North Carolina at Chapel Hill, Chapel Hill, United States
    For correspondence
    jesse_raab@med.unc.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6387-8994
  23. Bernard E Weissman

    Department of Pathology and Laboratory Medicine; Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, United States
    For correspondence
    weissman@med.unc.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1827-2309

Funding

National Institutes of Health (R01CA195670)

  • David G Huntsman
  • Jeffrey M Trent
  • Bernard E Weissman

National Institutes of Health (P30CA016086)

  • Joel S Parker

National Institutes of Health (T32ES007126)

  • Vinh Nguyen

Department of Defense (W81XWH-19-1-0423)

  • Jesse R Raab

National Institutes of Health (P51 OD 011092)

  • Larry S Sherman

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

Reviewing Editor

  1. Maureen E Murphy, The Wistar Institute, United States

Version history

  1. Received: May 19, 2020
  2. Accepted: December 22, 2020
  3. Accepted Manuscript published: December 23, 2020 (version 1)
  4. Version of Record published: January 18, 2021 (version 2)

Copyright

© 2020, Orlando 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

  • 3,597
    Page views
  • 294
    Downloads
  • 13
    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. Krystal Ann Orlando
  2. Amber K Douglas
  3. Aierken Abudu
  4. Yemin Wang
  5. Basile Tessier-Cloutier
  6. Weiping Su
  7. Alec Peters
  8. Larry S Sherman
  9. Rayvon Moore
  10. Vinh Nguyen
  11. Gian Luca Negri
  12. Shane Colborne
  13. Gregg B Morin
  14. Friedrich Kommoss
  15. Jessica D Lang
  16. William PD Hendricks
  17. Elizabeth A Raupach
  18. Patrick Pirrotte
  19. David G Huntsman
  20. Jeffrey M Trent
  21. Joel S Parker
  22. Jesse R Raab
  23. Bernard E Weissman
(2020)
Re-expression of SMARCA4/BRG1 in Small Cell Carcinoma of Ovary, Hypercalcemic Type (SCCOHT) promotes an epithelial-like gene signature through an AP-1-dependent mechanism
eLife 9:e59073.
https://doi.org/10.7554/eLife.59073

Further reading

    1. Cancer Biology
    Gehad Youssef, Luke Gammon ... Adrian Biddle
    Research Article

    Cancer stem cells (CSCs) undergo epithelial-mesenchymal transition (EMT) to drive metastatic dissemination in experimental cancer models. However, tumour cells undergoing EMT have not been observed disseminating into the tissue surrounding human tumour specimens, leaving the relevance to human cancer uncertain. We have previously identified both EpCAM and CD24 as CSC markers that, alongside the mesenchymal marker Vimentin, identify EMT CSCs in human oral cancer cell lines. This afforded the opportunity to investigate whether the combination of these three markers can identify disseminating EMT CSCs in actual human tumours. Examining disseminating tumour cells in over 12,000 imaging fields from 74 human oral tumours, we see a significant enrichment of EpCAM, CD24 and Vimentin co-stained cells disseminating beyond the tumour body in metastatic specimens. Through training an artificial neural network, these predict metastasis with high accuracy (cross-validated accuracy of 87-89%). In this study, we have observed single disseminating EMT CSCs in human oral cancer specimens, and these are highly predictive of metastatic disease.

    1. Cancer Biology
    2. Medicine
    Dingyu Rao, Hua Lu ... Defa Huang
    Research Article

    Esophageal cancer (EC) is a fatal digestive disease with a poor prognosis and frequent lymphatic metastases. Nevertheless, reliable biomarkers for EC diagnosis are currently unavailable. Accordingly, we have performed a comparative proteomics analysis on cancer and paracancer tissue-derived exosomes from eight pairs of EC patients using label-free quantification proteomics profiling and have analyzed the differentially expressed proteins through bioinformatics. Furthermore, nano-flow cytometry (NanoFCM) was used to validate the candidate proteins from plasma-derived exosomes in 122 EC patients. Of the 803 differentially expressed proteins discovered in cancer and paracancer tissue-derived exosomes, 686 were up-regulated and 117 were down-regulated. Intercellular adhesion molecule-1 (CD54) was identified as an up-regulated candidate for further investigation, and its high expression in cancer tissues of EC patients was validated using immunohistochemistry, real-time quantitative PCR (RT-qPCR), and western blot analyses. In addition, plasma-derived exosome NanoFCM data from 122 EC patients concurred with our proteomic analysis. The receiver operating characteristic (ROC) analysis demonstrated that the AUC, sensitivity, and specificity values for CD54 were 0.702, 66.13%, and 71.31%, respectively, for EC diagnosis. Small interference (si)RNA was employed to silence the CD54 gene in EC cells. A series of assays, including cell counting kit-8, adhesion, wound healing, and Matrigel invasion, were performed to investigate EC viability, adhesive, migratory, and invasive abilities, respectively. The results showed that CD54 promoted EC proliferation, migration, and invasion. Collectively, tissue-derived exosomal proteomics strongly demonstrates that CD54 is a promising biomarker for EC diagnosis and a key molecule for EC development.