Novel BRCA1 and BRCA2 Tumour Test as Basis for Treatment Decisions and Referral for Genetic Counselling of Patients with Ovarian Carcinomas.

Weren RD1, Mensenkamp AR1, Simons M2, Eijkelenboom A2, Sie AS1, Ouchene H1, van Asseldonk M2, Gomez-Garcia EB3, Blok MJ3, de Hullu JA4, Nelen MR1, Hoischen A1, Bulten J2, Tops BB2, Hoogerbrugge N1, Ligtenberg EM5,6.

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Abstract

With the recent introduction of Poly(ADP-ribose) polymerase (PARP) inhibitors, a promising novel therapy has become available for ovarian carcinoma patients with inactivating BRCA1 or BRCA2 mutations in their tumour. To select patients who may benefit from these treatments, assessment of the mutation status of BRCA1 and BRCA2 in the tumour 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. MLPA and 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 ovarian carcinomas, enabling improved hereditary cancer risk assessment and clinical treatment of ovarian cancer patients. This article is protected by copyright. All rights reserved.