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FRMD6 has tumor suppressor functions in prostate cancer

Oncogene volume 40, pages 763–776 (2021)Cite this article

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Abstract

Available tools for prostate cancer (PC) prognosis are suboptimal but may be improved by better knowledge about genes driving tumor aggressiveness. Here, we identified FRMD6 (FERM domain-containing protein 6) as an aberrantly hypermethylated and significantly downregulated gene in PC. Low FRMD6 expression was associated with postoperative biochemical recurrence in two large PC patient cohorts. In overexpression and CRISPR/Cas9 knockout experiments in PC cell lines, FRMD6 inhibited viability, proliferation, cell cycle progression, colony formation, 3D spheroid growth, and tumor xenograft growth in mice. Transcriptomic, proteomic, and phospho-proteomic profiling revealed enrichment of Hippo/YAP and c-MYC signaling upon FRMD6 knockout. Connectivity Map analysis and drug repurposing experiments identified pyroxamide as a new potential therapy for FRMD6 deficient PC cells. Finally, we established orthotropic Frmd6 and Pten, or Pten only (control) knockout in the ROSA26 mouse prostate. After 12 weeks, Frmd6/Pten double knockouts presented high-grade prostatic intraepithelial neoplasia (HG-PIN) and hyperproliferation, while Pten single-knockouts developed only regular PIN lesions and displayed lower proliferation. In conclusion, FRMD6 was identified as a novel tumor suppressor gene and prognostic biomarker candidate in PC.

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Fig. 1: FRMD6 in clinical samples.
Fig. 2: Functional studies in isogenic PC3, PC-3M, DU145, and DU145-MN1 cell lines.
Fig. 3: FRMD6 inhibits 3D spheroid and xenograft tumor growth.
Fig. 4: Pathway analysis of PC3_KO_FRMD6 vs. parental PC3 based on RNA-Seq data.
Fig. 5: Drug repurposing experiments and phospho-proteomics.
Fig. 6: Orthotropic gene editing in transgenic Rosa26-LSL-Cas9-eGFP mice.

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Acknowledgements

Prof. Raymond C. Bergan (Department of Medicine, Northwestern University, Chicago, USA) generously provided the PC-3M cell line. The DU145-MN1 cell line was kindly provided by Dr Volker Jung (Institute of Urology and Pediatric Urology, University of Saarland, Hamburg/Saar, Germany). We thank Dr Søren Vang, Dr Morten Muhlig, and Dr Michael Knudsen for bioinformatics support. The Danish Cancer Biobank (DCB) is acknowledged for biological material. Ros Eeles and Zsofia Kote-Karai are supported by NIHR support to the Biomedical Research Centre at The Institute of Cancer Research and Royal Marsden NHS Foundation Trust. Clara Cieza-Borella was supported by a donation from the Burstons and by research donations from the de Laszlo Foundation. This work was supported by The Danish Cancer Society (KDS), The Velux Foundation (KDS), The Novo Nordisk Foundation [NNF16OC0023048 (KDS); NNF16OC0022946 (MEJ); NNF14CC0001 (JVO)], The Independent Research Fund Denmark, The Harboe Foundation (JHJ), The Lundbeck Foundation [R231-2016-2682 (MEJ)], National Institute of Health Research (RE), The Biomedical Research Centre at the Institute of Cancer Research and Royal Marsden NHS Foundation Trust (RE), Richard and Debbi Burston, Nick Phillips and Bradshaw Foundation (CCB), and Aarhus University (JHJ/SHS).

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  1. These authors contributed equally: Jakob Haldrup, Siri. H. Strand

Authors and Affiliations

  1. Department of Molecular Medicine, Aarhus University Hospital, Aarhus, Denmark

    Jakob Haldrup, Siri H. Strand, Maibritt Norgaard, Stine Hedensted & Karina D. Sorensen

  2. Department of Clinical Medicine, Aarhus University, Aarhus, Denmark

    Jakob Haldrup, Siri H. Strand, Maria Riedel, Maibritt Norgaard, Stine Hedensted & Karina D. Sorensen

  3. Oncogenetics, Division of Genetics and Epidemiology, The Institute of Cancer Research, London, UK

    Clara Cieza-Borrella, Rosalind Eeles & Zsofia Kote-Jarai

  4. Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark

    Magnus E. Jakobsson & Jesper V. Olsen

  5. Department of Immunotechnology, Lund University, Medicon Village, 22100, Lund, Sweden

    Magnus E. Jakobsson

  6. Department of Biomedicine, Aarhus University, Aarhus, Denmark

    Frederik Dagnaes-Hansen & Martin Thomsen

  7. Institute of Pathology, Aarhus University Hospital, Aarhus, Denmark

    Benedicte Parm Ulhoi

  8. The Institute of Cancer Research and Royal Marsden NHS Foundation Trust, London, UK

    Rosalind Eeles

  9. Dept. of Urology, Aarhus University Hospital, Aarhus, Denmark

    Michael Borre

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Contributions

Conception and design: JHJ, SHS, and KDS. Development of methodology: JHJ, SHS, CCB, MT, and JVO. Acquisition of data JHJ, SHS, MR, SH, MN, MT, FDH, MB, and MEJ. Analysis and interpretation of data: JHJ, SHS, MEJ, MT, CCB, BU, and KDS. Writing of the manuscript: JHJ, SHS, and KDS. Revision and approval of the final manuscript: All authors. Study supervision: KDS, ZKJ, RE, and JVO.

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Correspondence to Karina D. Sorensen.

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Haldrup, J., Strand, S.H., Cieza-Borrella, C. et al. FRMD6 has tumor suppressor functions in prostate cancer. Oncogene 40, 763–776 (2021). https://doi.org/10.1038/s41388-020-01548-w

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