Advertisement

FANCM, RAD1, CHEK1 and TP53I3 act as BRCA-like tumor suppressors and are mutated in hereditary ovarian cancer

  • Jaime L. Lopes
    Affiliations
    Center for Molecular Medicine and Genetics and Department of Oncology, Wayne State University School of Medicine, 421 E. Canfield Street, Suite 3126, Detroit, MI 48201, USA

    Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA
    Search for articles by this author
  • Sophia Chaudhry
    Affiliations
    Center for Molecular Medicine and Genetics and Department of Oncology, Wayne State University School of Medicine, 421 E. Canfield Street, Suite 3126, Detroit, MI 48201, USA
    Search for articles by this author
  • Guilherme S. Lopes
    Affiliations
    Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA
    Search for articles by this author
  • Nancy K. Levin
    Affiliations
    Center for Molecular Medicine and Genetics and Department of Oncology, Wayne State University School of Medicine, 421 E. Canfield Street, Suite 3126, Detroit, MI 48201, USA
    Search for articles by this author
  • Michael A. Tainsky
    Correspondence
    Corresponding author.
    Affiliations
    Center for Molecular Medicine and Genetics and Department of Oncology, Wayne State University School of Medicine, 421 E. Canfield Street, Suite 3126, Detroit, MI 48201, USA
    Search for articles by this author

      Highlights

      • Deficiency of CHEK1, FANCM and TP53I3 led to reduced homologous recombination repair efficiency.
      • Deficiency of RAD1, CHEK1 or FANCM led to a decrease in cellular viability.
      • Deficiency of CHEK1, RAD1 or TP53I3 displayed increased sensitivity to cisplatin.
      • Deficiency of CHEK1, REC8 or RAD1 display increased sensitivity etoposide.
      • Results corroborate whole exome sequencing and bioinformatics analyses that identified FANCM, RAD1, CHEK1 and TP53I3, as putative germline risk loci in inherited breast and ovarian cancer risk.

      Abstract

      Although 25% of ovarian cancer cases are due to inherited factors, most of the genetic risk remains unexplained. We previously identified candidate genes through germline whole exome sequencing of BRCA1/BRCA2 negative ovarian cancer patients with familial risk. Here, we performed functional assessment to determine whether they act as BRCA-like tumor suppressors.
      Seven candidate risk genes were targeted by siRNA for mRNA depletion followed by functional assays for clonogenic survival, cytotoxicity to DNA damaging agents, and involvement in homologous recombination repair. BRCA1 and BRCA1 were targeted as standards for loss of function outcome.
      Knockdown of various candidate genes led to tumor suppressor phenotypes also observed in BRCA1/BRCA2 deficient cells. Deficiency of CHEK1, FANCM and TP53I3 led to reduced homologous recombination repair efficiency. Knockdown of RAD1, CHEK1 or FANCM led to a decrease in cellular viability and cells deficient in CHEK1, RAD1 or TP53I3 displayed increased sensitivity to cisplatin.
      Functional studies of candidate genes identified by whole exome sequencing complements bioinformatics techniques and aid the implication of novel risk loci. The results of this study suggest that genes found mutated in hereditary ovarian cancer, FANCM, RAD1, CHEK1 and TP53I3, act as BRCA-like tumor suppressors.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Cancer Genetics
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Pharoah P.D.P.
        • Tsai Y.Y.
        • Ramus S.J.
        • Phelan C.M.
        • Goode E.L.
        • Lawrenson K.
        • et al.
        GWAS meta-analysis and replication identifies three new susceptibility loci for ovarian cancer.
        Nat Genet. 2013; 45: 362-370
        • Manolio T.A.
        • Collins F.S.
        • Cox N.J.
        • Goldstein D.B.
        • Hindorff L.A.
        • Hunter D.J.
        • et al.
        Finding the missing heritability of complex diseases.
        Nature. 2009; 461: 747-753
        • Stafford J.L.
        • Chaudhry S.
        • Tainsky M.A.
        • Levin N.K.
        Whole exome sequencing: a necessary tool for the future of clinical cancer care.
        J Cancer Biol Res. 2017; 5
        • Stafford J.L.
        • Dyson G.
        • Levin N.K.
        • Chaudhry S.
        • Rosati R.
        • Kalpage H.
        • et al.
        Reanalysis of BRCA1/2 negative high risk ovarian cancer patients reveals novel germline risk loci and insights into missing heritability.
        PLoS One. 2017; 12e0178450
        • Bakker S.T.
        • van de Vrugt H.J.
        • Rooimans M.A.
        • Oostra A.B.
        • Steltenpool J.
        • Delzenne-Goette E.
        • et al.
        Fancm-deficient mice reveal unique features of Fanconi anemia complementation group M.
        Hum Mol Genet. 2009; 18: 3484-3495
        • Lam M.H.
        • Liu Q.
        • Elledge S.J.
        • Rosen J.M.
        Chk1 is haploinsufficient for multiple functions critical to tumor suppression.
        Cancer Cell. 2004; 6: 45-59
        • Shima N.
        • Alcaraz A.
        • Liachko I.
        • Buske T.R.
        • Andrews C.A.
        • Munroe R.J.
        • et al.
        A viable allele of Mcm4 causes chromosome instability and mammary adenocarcinomas in mice.
        Nat Genet. 2007; 39: 93-98
        • Han L.
        • Hu Z.
        • Liu Y.
        • Wang X.
        • Hopkins K.M.
        • Lieberman H.B.
        • et al.
        Mouse Rad1 deletion enhances susceptibility for skin tumor development.
        Mol Cancer. 2010; 9: 67
        • Xu H.
        • Beasley M.D.
        • Warren W.D.
        • van der Horst G.T.J.
        • McKay M.J.
        Absence of mouse REC8 cohesin promotes synapsis of sister chromatids in meiosis.
        Dev Cell. 2005; 8: 949-961
        • Maxwell C.A.
        • Benítez J.
        • Gómez-Baldó L.
        • Osorio A.
        • Bonifaci N.
        • Fernández-Ramires R.
        • et al.
        Interplay between BRCA1 and RHAMM regulates epithelial apicobasal polarization and may influence risk of breast cancer.
        PLoS Biol. 2011; 9: 1-18
        • Parvin J.
        • Chiba N.
        • Ransburgh D.
        Identifying the effects of BRCA1 mutations on homologous recombination using cells that express endogenous wild-type BRCA1.
        J Vis Exp. 2011; 48: 3-7
        • Towler W.I.
        • Zhang J.
        • Ransburgh D.J.R.
        • Toland A.E.
        • Ishioka C.
        • Chiba N.
        • et al.
        Analysis of BRCA1 variants in double-strand break repair by homologous recombination and single-strand annealing.
        Hum Mutat. 2013; 34: 439-445
        • Pierce A.J.
        • Johnson R.D.
        • Thompson L.H.
        • Jasin M.
        XRCC3 promotes homology-directed repair of DNA damage in mammalian cells.
        Genes Dev. 1999; 13: 2633-2638
        • Kim H.-Y.
        Statistical notes for clinical researchers: assessing normal distribution (2) using skewness and kurtosis.
        Restor Dent Endod. 2013; 38: 52
        • Ryland G.L.
        • Doyle M.A.
        • Goode D.
        • Boyle S.E.
        • Choong D.Y.H.
        • Rowley S.M.
        • et al.
        Loss of heterozygosity: what is it good for?.
        BMC Med Genom. 2015; 8: 1-12
        • Wang Y.
        • Huang B.
        • Edelmann L.
        • Kolodner R.D.
        • Edelmann W.
        Genes essential for cell viability that are linked to tumor suppressor genes play a role in cancer susceptibility.
        Cell Cycle. 2010; 9: 3144-3145
        • Pennington K.P.
        • Walsh T.
        • Harrell M.I.
        • Lee M.K.
        • Christopher C.
        • Rendi M.H.
        • et al.
        Germline and somatic mutations in homologous recombination genes predict platinum response and survival in ovarian, fallopian tube, and peritoneal carcinomas.
        Clini Cancer Res. 2014; 20: 764-775
        • Treszezamsky A.D.
        • Kachnic L.A.
        • Feng Z.
        • Zhang J.
        • Tokadjian C.
        • Powell S.N.
        BRCA1- and BRCA2-deficient cells are sensitive to etoposide-induced DNA double-strand breaks via topoisomerase II.
        Cancer Res. 2007; 67: 7078-7081
        • Farmer H.
        • McCabe N.
        • Lord C.J.
        • Tutt A.N.J.
        • Johnson D.A.
        • Richardson T.B.
        • et al.
        Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy.
        Nature. 2005; 434: 917-921
        • Yang J.
        • Bogni A.
        • Schuetz E.G.
        • Ratain M.
        • Eileen M.
        • Dolan H.
        • et al.
        Etoposide pathway.
        Pharmacogenet Genom. 2009; 19: 552-553
        • Bryant H.E.
        • Schultz N.
        • Thomas H.D.
        • Parker K.M.
        • Flower D.
        • Lopez E.
        • et al.
        Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase.
        Nature. 2005; 434: 913-917
        • Robson M.
        • Im S.-A.
        • Senkus E.
        • Xu B.
        • Domchek S.M.
        • Masuda N.
        • et al.
        Olaparib for metastatic breast cancer in patients with a germline BRCA mutation.
        N Engl J Med. 2017; 377: 523-533
        • Ledermann J.A.
        PARP inhibitors in ovarian cancer.
        Ann Oncol. 2016; 27 (no. December): i40-i44
        • Evans T.
        • Matulonis U.
        PARP inhibitors in ovarian cancer: evidence, experience and clinical potential.
        Ther Adv Med Oncol. 2017; 9: 253-267
        • Kim G.
        • Ison G.
        • McKee A.E.
        • Zhang H.
        • Tang S.
        • Gwise T.
        • et al.
        FDA approval summary: olaparib monotherapy in patients with deleterious germline BRCA-mutated advanced ovarian cancer treated with three or more lines of chemotherapy.
        Clin Cancer Res. 2015; 21: 4257-4261
        • Kaufman B.
        • Shapira-Frommer R.
        • Schmutzler R.K.
        • Audeh M.W.
        • Friedlander M.
        • Balmaña J.
        • et al.
        Olaparib monotherapy in patients with advanced cancer and a germline BRCA1/2 mutation.
        J Clin Oncol. 2015; 33: 244-250
        • Peterlongo P.
        • Catucci I.
        • Colombo M.
        • Caleca L.
        • Mucaki E.
        • Bogliolo M.
        • et al.
        FANCM c.5791C>T nonsense mutation (rs144567652) induces exon skipping, affects DNA repair activity and is a familial breast cancer risk factor.
        Hum Mol Genet. 2015; 24: 5345-5355
        • Dicks E.
        • Song H.
        • Ramus S.J.
        • Van Oudenhove E.
        • Tyrer P.
        • Intermaggio M.P.
        • et al.
        Germline whole exome sequencing and large-scale replication identifies FANCM as a likely high grade serous ovarian cancer susceptibility gene.
        Oncotarget. 2017; 8: 50930-50940
        • Liu Q.
        • Guntuku S.
        • Cui X.S.
        • Matsuoka S.
        • Cortez D.
        • Tamai K.
        • et al.
        Chk1 is an essential kinase that is regulated by Atr and required for the G2/M DNA damage checkpoint.
        Genes Dev. 2000; 14: 1448-1459
        • Al-Kaabi M.M.
        • Alshareeda A.T.
        • Jerjees D.A.
        • Muftah A.A.
        • Green A.R.
        • Alsubhi N.H.
        • et al.
        Checkpoint kinase1 (CHK1) is an important biomarker in breast cancer having a role in chemotherapy response.
        Br J Cancer. 2015; 112: 901-911
        • Mu K.
        • Li L.
        • Yang Q.
        • Zhang T.
        • Gao P.
        • Meng B.
        • et al.
        Detection of CHK1 and CCND1 gene copy number changes in breast cancer with dual-colour fluorescence in-situ hybridization.
        Histopathology. 2011; 58: 601-607
        • Ma J.
        • Kim E.M.
        • Haber J.E.
        • Lee S.E.
        Yeast Mre11 and Rad1 proteins define a Ku-independent mechanism to repair double-strand breaks lacking overlapping end sequences.
        Mol Cell Biol. 2003; 23: 8820-8828
        • Zhang W.
        • Luo J.
        • Chen F.
        • Yang F.
        • Song W.
        BRCA1 regulates PIG3-mediated apoptosis in a p53-dependent manner.
        Oncotarget. 2015; 6: 7606-7618
        • Yu J.
        • Liang Q.
        • Wang J.
        • Wang K.
        • Gao J.
        • Zhang J.
        • et al.
        REC8 functions as a tumor suppressor and is epigenetically downregulated in gastric cancer, especially in EBV-positive subtype.
        Oncogene. 2017; 36: 182-193
        • Liu D.
        • Shen X.
        • Zhu G.
        • Xing M.
        REC8 is a novel tumor suppressor gene epigenetically robustly targeted by the PI3K pathway in thyroid cancer.
        Oncotarget. 2015; 6: 39211-39224