title: Novel BRCA1 and BRCA2 Tumor Test as Basis for Treatment Decisions and Referral for Genetic Counselling of Patients with Ovarian Carcinomas.
abstract: With the recent introduction of Poly(ADP-ribose) polymerase inhibitors, a promising novel therapy has become available for ovarian carcinoma (OC) patients with inactivating BRCA1 or BRCA2 mutations in their tumor. To select patients who may benefit from these treatments, assessment of the mutation status of BRCA1 and BRCA2 in the tumor is required. For reliable evaluation of germline and somatic mutations in these genes in DNA derived from formalin-fixed, paraffin-embedded (FFPE) tissue, we have developed a single-molecule molecular inversion probe (smMIP)-based targeted next-generation sequencing (NGS) approach. Our smMIP-based NGS approach provides analysis of both strands of the open reading frame of BRCA1 and BRCA2, enabling the discrimination between real variants and formalin-induced artefacts. The single molecule tag enables compilation of unique reads leading to a high analytical sensitivity and enabling assessment of the reliability of mutation-negative results. Multiplex ligation-dependent probe amplification (MLPA) and Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) were used to detect exon deletions of BRCA1 and methylation of the BRCA1 promoter, respectively. Here, we show that this combined approach allows the rapid and reliable detection of both germline and somatic aberrations affecting BRCA1 and BRCA2 in DNA derived from FFPE OCs, enabling improved hereditary cancer risk assessment and clinical treatment of ovarian cancer patients.
authors: Department of Human Genetics, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Robbert D A
authors: RD
authors: Weren
authors: Department of Human Genetics, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Arjen R
authors: AR
authors: Mensenkamp
authors: Department of Pathology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Michiel
authors: M
authors: Simons
authors: Department of Pathology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Astrid
authors: A
authors: Eijkelenboom
authors: Department of Human Genetics, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Aisha S
authors: AS
authors: Sie
authors: Department of Human Genetics, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Hicham
authors: H
authors: Ouchene
authors: Department of Pathology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Monique
authors: M
authors: van Asseldonk
authors: Department of Clinical Genetics, GROW - School for Oncology & Developmental Biology, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors: Encarna B
authors: EB
authors: Gomez-Garcia
authors: Department of Clinical Genetics, GROW - School for Oncology & Developmental Biology, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors: Marinus J
authors: MJ
authors: Blok
authors: Department of Obstetrics & Gynaecology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Joanne A
authors: JA
authors: de Hullu
authors: Department of Human Genetics, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Marcel R
authors: MR
authors: Nelen
authors: Department of Human Genetics, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Alexander
authors: A
authors: Hoischen
authors: Department of Pathology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Johan
authors: J
authors: Bulten
authors: Department of Pathology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Bastiaan B J
authors: BB
authors: Tops
authors: Department of Human Genetics, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Nicoline
authors: N
authors: Hoogerbrugge
authors: Department of Human Genetics, Radboud University Medical Center, Nijmegen, The Netherlands.|Department of Pathology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: Marjolijn J L
authors: MJ
authors: Ligtenberg
mesh_terms: D000483:Alleles; D019943:Amino Acid Substitution; D000066491:Clinical Decision-Making; D056915:DNA Copy Number Variations; D004252:DNA Mutational Analysis; D019468:Disease Management; D005260:Female; D019398:Genes, BRCA1; D024522:Genes, BRCA2; D056726:Genetic Association Studies; D005817:Genetic Counseling; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000379:methods / Q000592:standards; D005838:Genotype; D018095:Germ-Line Mutation; D006801:Humans; D019656:Loss of Heterozygosity; D010051:Ovarian Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000628:therapy; D015203:Reproducibility of Results
keywords: BRCA testing; BRCA1; BRCA2; PARP-inhibitor; cancer predisposition; ovarian cancer; personalized medicine; single molecule molecular inversion probes