Advertisement

Germline investigation in male breast cancer of DNA repair genes by next-generation sequencing

  • R. Scarpitta
  • I. Zanna
  • P. Aretini
  • G. Gambino
  • C. Scatena
  • B. Mei
  • M. Ghilli
  • E. Rossetti
  • M. Roncella
  • C. Congregati
  • F. Bonci
  • A. G. Naccarato
  • D. Palli
  • M. A. CaligoEmail author
Preclinical Study

Abstract

Purpose

In order to better define the breast cancer (BC) genetic risk factors in men, a germline investigation was carried out on 81 Male BC cases by screening the 24 genes involved in BC predisposition, genome stability maintenance and DNA repair mechanisms by next-generation sequencing.

Methods

Germline DNAs were tested in a custom multi-gene panel focused on all coding exons and exon–intron boundaries of 24 selected genes using two amplicon-based assays on PGM-Ion Torrent (ThermoFisher Scientific) and MiSeq (Illumina) platforms. All variants were recorded and classified by using a custom pipeline.

Results

Clinical pathological data and the family history of 81 Male BC cases were gathered and analysed, revealing the average age of onset to be 61.3 years old and that in 35 cases there was a family history of BC. Our genetic screening allowed us to identify a germline mutation in 22 patients (23%) in 4 genes: BRCA2, BRIP1, MUTYH and PMS2. Moreover, 12 variants of unknown clinical significance (VUS) in 9 genes (BARD1, BRCA1, BRIP1, CHEK2, ERCC1, NBN, PALB2, PMS1, RAD50) were predicted as potentially pathogenic by in silico analysis bringing the mutation detection rate up to 40%.

Conclusion

As expected, a positive family history is a strong predictor of germline BRCA2 mutations in male BC. Understanding the potential pathogenicity of VUS represents an extremely urgent need for the management of BC risk in Male BC cases and their own families.

Keywords

Male breast cancer Next-generation sequencing DNA repair genes Familial breast cancer Breast cancer risk in men 

Notes

Acknowledgements

The authors would like to acknowledge all the patients that were involved in this study.

Funding

This study was supported by Grants from the Istituto Toscano Tumori (ITT) Grant 2010 and from the Fondazione Pisa Grant 2016 (prog.127/16 and prog.148/16), and from research funding Susan G. Komen Italia onlus 2017.

Compliance with ethical standards

Conflict of interest

M.A. Caligo was supported by Grant 2016 (prog.127/16) from the Fondazione Pisa and by research funding 2017 from the Susan G. Komen Italia onlus. A. G. Naccarato was supported by Grant 2016 (prog.148/16) from the Fondazione Pisa. D. Palli was supported by Grant 2010 from the Istituto Toscano Tumori (ITT). All authors declare that there are no conflicts of interest.

Ethical approval

All the procedures performed in studies involving human participants were in accordance with the Ethical Standards of the Institutional and/or National Research Committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

10549_2019_5429_MOESM1_ESM.docx (45 kb)
Supplementary material 1 (DOCX 45 kb)

References

  1. 1.
    Siegel R, Naishadham D, Jemal A (2012) Cancer statistics, 2012. CA Cancer J Clin 62:10–29.  https://doi.org/10.3322/caac.20138 CrossRefGoogle Scholar
  2. 2.
    Ottini L, Palli D, Rizzo S et al (2010) Male breast cancer. Crit Rev Oncol Hematol 73:141–155.  https://doi.org/10.1016/j.critrevonc.2009.04.003 CrossRefGoogle Scholar
  3. 3.
    Giordano SH, Cohen DS, Buzdar AU et al (2004) Breast carcinoma in men: a population-based study. Cancer 101:51–57.  https://doi.org/10.1002/cncr.20312 CrossRefGoogle Scholar
  4. 4.
    Severson TM, Zwart W (2017) A review of estrogen receptor/androgen receptor genomics in male breast cancer. Endocr Relat Cancer 24:R27–R34.  https://doi.org/10.1530/ERC-16-0225 CrossRefGoogle Scholar
  5. 5.
    Howlader N, Noone AM, Krapcho M (2013) Lifetime risk of developing or dying from cancer. SEER Cancer Statistics Review 1975–2011Google Scholar
  6. 6.
    Mazzanti CM, Lessi F, Armogida I et al (2015) Human saliva as route of inter-human infection for mouse mammary tumor virus. Oncotarget.  https://doi.org/10.18632/oncotarget.4567 Google Scholar
  7. 7.
    Miki Y, Swensen J, Shattuck-Eidens D et al (1994) A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1. Science 266:66–71.  https://doi.org/10.1126/science.7545954 CrossRefGoogle Scholar
  8. 8.
    Stratton MR, Ford D, Neuhausen S et al (1994) Familial male breast cancer is not linked to the BRCA1 locus on chromosome 17q. Nat Genet 7:103–107.  https://doi.org/10.1038/ng0594-103 CrossRefGoogle Scholar
  9. 9.
    Tai YC, Domchek S, Parmigiani G, Chen S (2007) Breast cancer risk among male BRCA1 and BRCA2 mutation carriers. J Natl Cancer Inst 99:1811–1814.  https://doi.org/10.1093/jnci/djm203 CrossRefGoogle Scholar
  10. 10.
    Rizzolo P, Silvestri V, Tommasi S et al (2013) Male breast cancer: genetics, epigenetics, and ethical aspects. Ann Oncol 24:viii75–viii82.  https://doi.org/10.1093/annonc/mdt316 CrossRefGoogle Scholar
  11. 11.
    Frank TS, Deffenbaugh AM, Reid JE et al (2002) Clinical characteristics of individuals with germline mutations in BRCA1 and BRCA2: analysis of 10,000 individuals. J Clin Oncol 20:1480–1490CrossRefGoogle Scholar
  12. 12.
    Ottini L, Silvestri V, Rizzolo P et al (2012) Clinical and pathologic characteristics of BRCA-positive and BRCA-negative male breast cancer patients: results from a collaborative multicenter study in Italy. Breast Cancer Res Treat 134:411–418.  https://doi.org/10.1007/s10549-012-2062-0 CrossRefGoogle Scholar
  13. 13.
    Silvestri V, Rizzolo P, Zanna I et al (2010) PALB2 mutations in male breast cancer: a population-based study in central Italy. Breast Cancer Res Treat 122:299–301.  https://doi.org/10.1007/s10549-010-0797-z CrossRefGoogle Scholar
  14. 14.
    Pritzlaff M, Summerour P, McFarland R et al (2017) Male breast cancer in a multi-gene panel testing cohort: insights and unexpected results. Breast Cancer Res Treat 161:575–586.  https://doi.org/10.1007/s10549-016-4085-4 CrossRefGoogle Scholar
  15. 15.
    Fostira F, Saloustros E, Apostolou P et al (2018) Germline deleterious mutations in genes other than BRCA2 are infrequent in male breast cancer. Breast Cancer Res Treat 169:105–113.  https://doi.org/10.1007/s10549-018-4661-x CrossRefGoogle Scholar
  16. 16.
    Nielsen FC, van Overeem Hansen T, Sørensen CS (2016) Hereditary breast and ovarian cancer: new genes in confined pathways. Nat Rev Cancer 16:599–612.  https://doi.org/10.1038/nrc.2016.72 CrossRefGoogle Scholar
  17. 17.
    Tung N, Battelli C, Allen B et al (2015) Frequency of mutations in individuals with breast cancer referred for BRCA1 and BRCA2 testing using next-generation sequencing with a 25-gene panel: mutations in BRCA1/2-tested patients. Cancer 121:25–33.  https://doi.org/10.1002/cncr.29010 CrossRefGoogle Scholar
  18. 18.
    Easton DF, Pharoah PDP, Antoniou AC et al (2015) Gene-panel sequencing and the prediction of breast-cancer risk. N Engl J Med 372:2243–2257.  https://doi.org/10.1056/NEJMsr1501341 CrossRefGoogle Scholar
  19. 19.
    Rizzolo P, Zelli V, Silvestri V et al (2019) Insight into genetic susceptibility to male breast cancer by multigene panel testing: results from a multicenter study in Italy: multigene panel testing for male breast cancer predisposition. Int J Cancer.  https://doi.org/10.1002/ijc.32106 Google Scholar
  20. 20.
    Plon SE, Eccles DM, Easton D et al (2008) Sequence variant classification and reporting: recommendations for improving the interpretation of cancer susceptibility genetic test results. Hum Mutat 29:1282–1291.  https://doi.org/10.1002/humu.20880 CrossRefGoogle Scholar
  21. 21.
    Richards S, on behalf of the ACMG Laboratory Quality Assurance Committee, Aziz N et al (2015) Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med 17:405–423.  https://doi.org/10.1038/gim.2015.30
  22. 22.
    Korde LA, Zujewski JA, Kamin L et al (2010) Multidisciplinary Meeting on Male Breast Cancer: summary and research recommendations. J Clin Oncol 28:2114–2122.  https://doi.org/10.1200/JCO.2009.25.5729 CrossRefGoogle Scholar
  23. 23.
    van der Hout AH, van den Ouweland AMW, van der Luijt RB et al (2006) A DGGE system for comprehensive mutation screening of BRCA1 and BRCA2: application in a Dutch cancer clinic setting. Hum Mutat 27:654–666.  https://doi.org/10.1002/humu.20340 CrossRefGoogle Scholar
  24. 24.
    Marroni F, Cipollini G, Peissel B et al (2008) Reconstructing the genealogy of a BRCA1 founder mutation by phylogenetic analysis. Ann Hum Genet 72:310–318.  https://doi.org/10.1111/j.1469-1809.2007.00420.x CrossRefGoogle Scholar
  25. 25.
    Cipollini G (2004) Genetic alterations in hereditary breast cancer. Ann Oncol 15:i7–i13.  https://doi.org/10.1093/annonc/mdh651 CrossRefGoogle Scholar
  26. 26.
    Malacrida S, Agata S, Callegaro M et al (2008) BRCA1 p.Val1688del is a deleterious mutation that recurs in breast and ovarian cancer families from northeast Italy. J Clin Oncol 26:26–31.  https://doi.org/10.1200/JCO.2007.13.2118 CrossRefGoogle Scholar
  27. 27.
    Palmieri G (2002) BRCA1 and BRCA2 germline mutations in Sardinian breast cancer families and their implications for genetic counseling. Ann Oncol 13:1899–1907.  https://doi.org/10.1093/annonc/mdf326 CrossRefGoogle Scholar
  28. 28.
    Wilson BT, Douglas SF, Polvikoski T (2010) Astrocytoma in a breast cancer lineage: part of the BRCA2 phenotype? J Clin Oncol 28:e596–e598.  https://doi.org/10.1200/JCO.2010.28.9173 CrossRefGoogle Scholar
  29. 29.
    Offit K, Levran O, Mullaney B et al (2003) Shared genetic susceptibility to breast cancer, brain tumors, and Fanconi anemia. J Natl Cancer Inst 95:1548–1551.  https://doi.org/10.1093/jnci/djg072 CrossRefGoogle Scholar
  30. 30.
    Reid S (2005) Biallelic BRCA2 mutations are associated with multiple malignancies in childhood including familial Wilms tumour. J Med Genet 42:147–151.  https://doi.org/10.1136/jmg.2004.022673 CrossRefGoogle Scholar
  31. 31.
    Dodgshun AJ, Sexton-Oates A, Saffery R, Sullivan MJ (2016) Biallelic FANCD1/BRCA2 mutations predisposing to glioblastoma multiforme with multiple oncogenic amplifications. Cancer Genet 209:53–56.  https://doi.org/10.1016/j.cancergen.2015.11.005 CrossRefGoogle Scholar
  32. 32.
    Gaildrat P, Krieger S, Di Giacomo D et al (2012) Multiple sequence variants of BRCA2 exon 7 alter splicing regulation. J Med Genet 49:609–617.  https://doi.org/10.1136/jmedgenet-2012-100965 CrossRefGoogle Scholar
  33. 33.
    Colombo M, Ripamonti CB, Pensotti V et al (2009) An unusual BRCA2 allele carrying two splice site mutations. Ann Oncol 20:1143–1144.  https://doi.org/10.1093/annonc/mdp241 CrossRefGoogle Scholar
  34. 34.
    Pensabene M, Spagnoletti I, Capuano I et al (2009) Two mutations of BRCA2 gene at exon and splicing site in a woman who underwent oncogenetic counseling. Ann Oncol 20:874–878.  https://doi.org/10.1093/annonc/mdn724 CrossRefGoogle Scholar
  35. 35.
    Fackenthal JD, Yoshimatsu T, Zhang B et al (2016) Naturally occurring BRCA2 alternative mRNA splicing events in clinically relevant samples. J Med Genet 53:548–558.  https://doi.org/10.1136/jmedgenet-2015-103570 CrossRefGoogle Scholar
  36. 36.
    Gambino G, Tancredi M, Falaschi E et al (2015) Characterization of three alternative transcripts of the BRCA1 gene in patients with breast cancer and a family history of breast and/or ovarian cancer who tested negative for pathogenic mutations. Int J Mol Med 35:950–956.  https://doi.org/10.3892/ijmm.2015.2103 CrossRefGoogle Scholar
  37. 37.
    Caputo SM, Léone M, Damiola F et al (2018) Full in-frame exon 3 skipping of brca2 confers high risk of breast and/or ovarian cancer. Oncotarget.  https://doi.org/10.18632/oncotarget.24671 Google Scholar
  38. 38.
    Bonatti F, Pepe C, Tancredi M et al (2006) RNA-based analysis of BRCA1 and BRCA2 gene alterations. Cancer Genet Cytogenet 170:93–101.  https://doi.org/10.1016/j.cancergencyto.2006.05.005 CrossRefGoogle Scholar
  39. 39.
    Acedo A, Hernández-Moro C, Curiel-García Á et al (2015) Functional classification of BRCA2 DNA variants by splicing assays in a large minigene with 9 exons. Hum Mutat 36:210–221.  https://doi.org/10.1002/humu.22725 CrossRefGoogle Scholar
  40. 40.
    Cantor SB, Guillemette S (2011) Hereditary breast cancer and the BRCA1-associated FANCJ/BACH1/BRIP1. Future Oncol 7:253–261.  https://doi.org/10.2217/fon.10.191 CrossRefGoogle Scholar
  41. 41.
    Spugnesi L, Gabriele M, Scarpitta R et al (2016) Germline mutations in DNA repair genes may predict neoadjuvant therapy response in triple negative breast patients: DNA repair mutations in neoadjuvant TNBCS. Genes Chromosomes Cancer 55:915–924.  https://doi.org/10.1002/gcc.22389 CrossRefGoogle Scholar
  42. 42.
    De Nicolo A, Tancredi M, Lombardi G et al (2008) A novel breast cancer-associated BRIP1 (FANCJ/BACH1) germ-line mutation impairs protein stability and function. Clin Cancer Res 14:4672–4680.  https://doi.org/10.1158/1078-0432.CCR-08-0087 CrossRefGoogle Scholar
  43. 43.
    Wasielewski M, Out AA, Vermeulen J et al (2010) Increased MUTYH mutation frequency among Dutch families with breast cancer and colorectal cancer. Breast Cancer Res Treat 124:635–641.  https://doi.org/10.1007/s10549-010-0801-7 CrossRefGoogle Scholar
  44. 44.
    Rizzolo P, Silvestri V, Bucalo A et al (2018) Contribution of MUTYH variants to male breast cancer risk: results from a multicenter study in Italy. Front Oncol.  https://doi.org/10.3389/fonc.2018.00583 Google Scholar
  45. 45.
    ten Broeke SW, Brohet RM, Tops CM et al (2015) Lynch syndrome caused by germline PMS2 mutations: delineating the cancer risk. J Clin Oncol 33:319–325.  https://doi.org/10.1200/JCO.2014.57.8088 CrossRefGoogle Scholar
  46. 46.
    Maresca L, Spugnesi L, Lodovichi S et al (2015) MSH2 role in BRCA1-driven tumorigenesis: a preliminary study in yeast and in human tumors from BRCA1-VUS carriers. Eur J Med Genet 58:531–539.  https://doi.org/10.1016/j.ejmg.2015.09.005 CrossRefGoogle Scholar
  47. 47.
    Boyd J, Rhei E, Federici MG et al (1999) Male breast cancer in the hereditary nonpolyposis colorectal cancer syndrome. Breast Cancer Res Treat 53:87–91CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • R. Scarpitta
    • 1
  • I. Zanna
    • 2
  • P. Aretini
    • 3
  • G. Gambino
    • 1
  • C. Scatena
    • 4
  • B. Mei
    • 1
  • M. Ghilli
    • 5
  • E. Rossetti
    • 5
  • M. Roncella
    • 5
  • C. Congregati
    • 6
  • F. Bonci
    • 7
  • A. G. Naccarato
    • 4
  • D. Palli
    • 2
  • M. A. Caligo
    • 1
    Email author
  1. 1.Section of Genetic OncologyUniversity HospitalPisaItaly
  2. 2.Cancer Risk Factors and Lifestyle Epidemiology UnitInstitute for Cancer Research, Prevention and Clinical Network (ISPRO)FlorenceItaly
  3. 3.Section of Cancer GenomicsFondazione Pisana per la ScienzaPisaItaly
  4. 4.Division of Pathology, Department of Translational Research and New Technologies in Medicine and SurgeryUniversity of PisaPisaItaly
  5. 5.Breast Cancer CenterUniversity HospitalPisaItaly
  6. 6.Division of Internal MedicineUniversity HospitalPisaItaly
  7. 7.Unit of Medical Oncology 2University HospitalPisaItaly

Personalised recommendations