title: Investigating the effect of 28 BRCA1 and BRCA2 mutations on their related transcribed mRNA.
abstract: Germline inactivating mutations in the BRCA1 and BRCA2 genes are responsible for hereditary breast and ovarian cancer syndrome (HBOCS). Genetic testing of these genes identifies a significant proportion of variants of uncertain significance (VUS). Elucidation of the clinical impact of these variants is an important challenge in genetic diagnostics and counseling. In this study, we assess the RNA effect of 28 BRCA1 and BRCA2 VUS identified in our set of HBOCS families with the aim of gaining insight into their clinical relevance. mRNA was extracted from VUS carriers and controls lymphocytes cultured for 5-6 days and treated with puromycin. RNA was reverse transcribed to perform transcriptional analysis for the study of splicing aberrations. In silico prediction tools were used to select those variants most likely to affect the RNA splicing process. Six out of the 28 variants analyzed showed an aberrant splicing pattern and could therefore be classified as probably pathogenic mutations. Reclassification of VUS improves the genetic counseling and clinical surveillance of carriers of these mutations and highlights the importance of RNA studies in routine diagnostic laboratories.
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Quiles
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Mireia
authors.initials: M
authors.lastname: Menéndez
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Tornero
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Jesús
authors.initials: J
authors.lastname: del Valle
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Àlex
authors.initials: À
authors.lastname: Teulé
authors.affiliation: Molecular Biology Laboratory, Service of Clinical Analysis, University and Polytechnic Hospital La Fe, Valencia, Spain.
authors.forename: Sarai
authors.initials: S
authors.lastname: Palanca
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Angel
authors.initials: A
authors.lastname: Izquierdo
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Carolina
authors.initials: C
authors.lastname: Gómez
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Olga
authors.initials: O
authors.lastname: Campos
authors.affiliation: Genètica Molecular, Laboratorio Echevarne, Barcelona, Spain.
authors.forename: Raül
authors.initials: R
authors.lastname: Santamaria
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Joan
authors.initials: J
authors.lastname: Brunet
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Capellá
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain.
authors.forename: Lídia
authors.initials: L
authors.lastname: Feliubadaló
authors.affiliation: Hereditary Cancer Program, Molecular Diagnostics Unit, Catalan Institute of Oncology (ICO-IDIBELL), Hospital Duran i Reynals, Gran Via 199, L'Hospitalet de Llobregat, 08908, Barcelona, Spain. conxi.lazaro@gmail.com.
authors.forename: Conxi
authors.initials: C
authors.lastname: Lázaro
mesh_terms: D019313:BRCA1 Protein / Q000235:genetics*; D024682:BRCA2 Protein / Q000235:genetics*; D005260:Female; D018095:Germ-Line Mutation / Q000235:genetics*; D061325:Hereditary Breast and Ovarian Cancer Syndrome / Q000235:genetics*; D006801:Humans; D012326:RNA Splicing / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics
chemical_list: D019313:BRCA1 Protein; D024682:BRCA2 Protein; C551750:BRCA2 protein, human; D012333:RNA, Messenger
keywords: BRCA1; BRCA2; RNA; Splicing; VUS