Angelo Minucci1, Maurizio Lalle2, Rossella De Leo3, Giorgia Mazzuccato4, Giovanni Scambia5, Andrea Urbani6, Anna Fagotti5, Paola Concolino4, Ettore Capoluongo6. 1. Dipartimento di Diagnostica di Laboratorio, Fondazione Policlinico Universitario A. Gemelli IRCCS, Roma, Italy. Electronic address: angelo.minucci@virgilio.it. 2. UOC Oncologia, Azienda Ospedaliera S. Giovanni - Addolorata, Roma, Italy. 3. Dipartimento di Ostetricia e Ginecologia, Divisione di Ginecologia Oncologica, Fondazione Policlinico Universitario A. Gemelli IRCCS, Roma, Italy. 4. Dipartimento di Diagnostica di Laboratorio, Fondazione Policlinico Universitario A. Gemelli IRCCS, Roma, Italy. 5. Dipartimento di Ostetricia e Ginecologia, Divisione di Ginecologia Oncologica, Fondazione Policlinico Universitario A. Gemelli IRCCS, Roma, Italy; Università Cattolica del Sacro Cuore, Roma, Italy. 6. Dipartimento di Diagnostica di Laboratorio, Fondazione Policlinico Universitario A. Gemelli IRCCS, Roma, Italy; Università Cattolica del Sacro Cuore, Roma, Italy.
Abstract
OBJECTIVES: In silico splicing analysis, a mini-gene assay and splicing data, obtained using RNA from blood samples, have shown that the BRCA1 c.5332G > A variant induces exon 21 skipping. However, despite these evidences, up to date, this variant is unclassified. The aim of this study is to provide further molecular and clinical evidence for the BRCA1 c.5332G > A variant in a patient with high grade serous ovarian carcinoma (HGSOC) to allow a definitive classification of this variant. DESIGN AND METHOD: The effect of the BRCA1 c.5332G > A variant on RNA splicing was evaluated by amplifying regions of BRCA1 from the cDNA of the patient. Loss of heterozygosity (LOH) in tumor tissue was also investigated. RESULTS: The c.5332G > A allele causes significantly aberrant splicing of the BRCA1 exon 21. Evaluation of the c.5332A allele in tumor tissue highlights a possible loss of heterozygosity, supporting her pathogenic effect. CONCLUSIONS: Our results regarding the c.5332G > A variant confirm that it contributed to predisposition and onset of ovarian carcinoma in the patient. We propose to classify this variant as 'likely-pathogenic' (class IV).
OBJECTIVES: In silico splicing analysis, a mini-gene assay and splicing data, obtained using RNA from blood samples, have shown that the BRCA1 c.5332G > A variant induces exon 21 skipping. However, despite these evidences, up to date, this variant is unclassified. The aim of this study is to provide further molecular and clinical evidence for the BRCA1 c.5332G > A variant in a patient with high grade serous ovarian carcinoma (HGSOC) to allow a definitive classification of this variant. DESIGN AND METHOD: The effect of the BRCA1 c.5332G > A variant on RNA splicing was evaluated by amplifying regions of BRCA1 from the cDNA of the patient. Loss of heterozygosity (LOH) in tumor tissue was also investigated. RESULTS: The c.5332G > A allele causes significantly aberrant splicing of the BRCA1 exon 21. Evaluation of the c.5332A allele in tumor tissue highlights a possible loss of heterozygosity, supporting her pathogenic effect. CONCLUSIONS: Our results regarding the c.5332G > A variant confirm that it contributed to predisposition and onset of ovarian carcinoma in the patient. We propose to classify this variant as 'likely-pathogenic' (class IV).
Authors: Angelo Minucci; Giorgia Mazzuccato; Marco D'Indinosante; Lucia Di Nardo; Paola Concolino; Maria De Bonis; Andrea Urbani; Giovanni Scambia; Anna Fagotti; Ettore Capoluongo Journal: Mol Biol Rep Date: 2019-12-12 Impact factor: 2.316