A multi-gene panel beyond BRCA1/BRCA2 to identify new breast cancer-predisposing mutations by a picodroplet PCR followed by a next-generation sequencing strategy: a pilot study

Marcella Nunziato, Maria Valeria Esposito, Flavio Starnone, Maria Angela Diroma, Alessandra Calabrese, Valentina Del Monaco, Pasqualina Buono, Giuseppe Frasci, Gerardo Botti, Massimiliano D'Aiuto, Francesco Salvatore, Valeria D'Argenio

Research output: Contribution to journalArticle

Abstract

By analyzing multiple gene panels, next-generation sequencing is more effective than conventional procedures in identifying disease-related mutations that are useful for clinical decision-making. Here, we aimed to test the efficacy of an 84 genes customized-panel in BRCA1 and BRCA2 mutation-negative patients. Twenty-four patients were enrolled in this study. DNA libraries were prepared using a picodroplet PCR-based approach and sequenced with the MiSeq System. Highly putative pathogenic mutations were identified in genes other than the commonly tested BRCA1/2: 2 pathogenic mutations one in TP53 and one in MUTYH; 2 missense variants in MSH6 and ATM, respectively; 2 frameshift variants in KLLN, and ATAD2, respectively; an intronic variant in ANPEP, and 3 not functionally known variants (a frameshift variant in ATM a nonsense variant in ATM and a missense variant in NFE2L2). Our results show that this molecular screening will increase diagnostic sensitivity leading to a better risk assessment in breast cancer patients and their families. This strategy could also reveal genes that have a higher penetrance for breast and ovarian cancers by matching gene mutation with familial and clinical data, thereby increasing information about hereditary breast and ovarian cancer genetics and improving cancer prevention measures or therapeutic approaches.

Original languageEnglish
Pages (from-to)154-162
Number of pages9
JournalAnalytica Chimica Acta
Volume1046
DOIs
Publication statusPublished - Jan 10 2019

Keywords

  • Adult
  • Aged
  • Aged, 80 and over
  • BRCA1 Protein/genetics
  • BRCA2 Protein/genetics
  • Breast Neoplasms/genetics
  • DNA Mutational Analysis/methods
  • Female
  • Humans
  • Male
  • Middle Aged
  • Mutation
  • Particle Size
  • Pedigree
  • Pilot Projects
  • Polymerase Chain Reaction

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