Optimizing the identification of risk-relevant mutations by multigene panel testing in selected hereditary breast/ovarian cancer families

Cancer Med. 2018 Jan;7(1):46-55. doi: 10.1002/cam4.1251. Epub 2017 Dec 22.

Abstract

The introduction of multigene panel testing for hereditary breast/ovarian cancer screening has greatly improved efficiency, speed, and costs. However, its clinical utility is still debated, mostly due to the lack of conclusive evidences on the impact of newly discovered genetic variants on cancer risk and lack of evidence-based guidelines for the clinical management of their carriers. In this pilot study, we aimed to test whether a systematic and multiparametric characterization of newly discovered mutations could enhance the clinical utility of multigene panel sequencing. Out of a pool of 367 breast/ovarian cancer families Sanger-sequenced for BRCA1 and BRCA2 gene mutations, we selected a cohort of 20 BRCA1/2-negative families to be subjected to the BROCA-Cancer Risk Panel massive parallel sequencing. As a strategy for the systematic characterization of newly discovered genetic variants, we collected blood and cancer tissue samples and established lymphoblastoid cell lines from all available individuals in these families, to perform segregation analysis, loss-of-heterozygosity and further molecular studies. We identified loss-of-function mutations in 6 out 20 high-risk families, 5 of which occurred on BRCA1, CHEK2 and ATM and are esteemed to be risk-relevant. In contrast, a novel RAD50 truncating mutation is most likely unrelated to breast cancer. Our data suggest that integrating multigene panel testing with a pre-organized, multiparametric characterization of newly discovered genetic variants improves the identification of risk-relevant alleles impacting on the clinical management of their carriers.

Keywords: ATM; CHEK2; NGS; BRCAPro5; hereditary breast cancer.

MeSH terms

  • Acid Anhydride Hydrolases
  • Adult
  • Ataxia Telangiectasia Mutated Proteins / genetics
  • BRCA1 Protein / genetics
  • BRCA2 Protein / genetics
  • Checkpoint Kinase 2 / genetics
  • Cohort Studies
  • DNA Mutational Analysis / methods
  • DNA Repair Enzymes / genetics
  • DNA-Binding Proteins / genetics
  • Female
  • Genetic Predisposition to Disease*
  • Genetic Testing / methods*
  • Hereditary Breast and Ovarian Cancer Syndrome / genetics*
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Loss of Function Mutation
  • Middle Aged
  • Pilot Projects

Substances

  • BRCA1 Protein
  • BRCA1 protein, human
  • BRCA2 Protein
  • BRCA2 protein, human
  • DNA-Binding Proteins
  • Checkpoint Kinase 2
  • ATM protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • CHEK2 protein, human
  • Acid Anhydride Hydrolases
  • RAD50 protein, human
  • DNA Repair Enzymes