TY - JOUR
T1 - 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
T2 - a pilot study
AU - Nunziato, Marcella
AU - Esposito, Maria Valeria
AU - Starnone, Flavio
AU - Diroma, Maria Angela
AU - Calabrese, Alessandra
AU - Del Monaco, Valentina
AU - Buono, Pasqualina
AU - Frasci, Giuseppe
AU - Botti, Gerardo
AU - D'Aiuto, Massimiliano
AU - Salvatore, Francesco
AU - D'Argenio, Valeria
N1 - Copyright © 2018. Published by Elsevier B.V.
PY - 2019/1/10
Y1 - 2019/1/10
N2 - 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.
AB - 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.
KW - Adult
KW - Aged
KW - Aged, 80 and over
KW - BRCA1 Protein/genetics
KW - BRCA2 Protein/genetics
KW - Breast Neoplasms/genetics
KW - DNA Mutational Analysis/methods
KW - Female
KW - Humans
KW - Male
KW - Middle Aged
KW - Mutation
KW - Particle Size
KW - Pedigree
KW - Pilot Projects
KW - Polymerase Chain Reaction
U2 - 10.1016/j.aca.2018.09.032
DO - 10.1016/j.aca.2018.09.032
M3 - Article
C2 - 30482293
VL - 1046
SP - 154
EP - 162
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
ER -