A High-Throughput Functional Complementation Assay for Classification of<i>BRCA1</i>Missense Variants
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- Peter Bouwman
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Hanneke van der Gulden
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Ingrid van der Heijden
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Rinske Drost
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Christiaan N. Klijn
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Pramudita Prasetyanti
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Mark Pieterse
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Ellen Wientjens
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Jost Seibler
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Frans B.L. Hogervorst
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
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- Jos Jonkers
- Authors' Affiliations: 1Division of Molecular Pathology and Cancer Genomics Centre and 2Department of Pathology, The Netherlands Cancer Institute Amsterdam, The Netherlands; and 3TaconicArtemis GmbH, Cologne, Germany
説明
<jats:title>Abstract</jats:title><jats:p>Mutations in BRCA1 and BRCA2 account for the majority of hereditary breast and ovarian cancers, and therefore sequence analysis of both genes is routinely conducted in patients with early-onset breast cancer. Besides mutations that clearly abolish protein function or are known to increase cancer risk, a large number of sequence variants of uncertain significance (VUS) have been identified. Although several functional assays for BRCA1 VUSs have been described, thus far it has not been possible to conduct a high-throughput analysis in the context of the full-length protein. We have developed a relatively fast and easy cDNA-based functional assay to classify BRCA1 VUSs based on their ability to functionally complement BRCA1-deficient mouse embryonic stem cells. Using this assay, we have analyzed 74 unclassified BRCA1 missense mutants for which all predicted pathogenic variants are confined to the BRCA1 RING and BRCT domains.</jats:p><jats:p>Significance: BRCA1 VUSs are frequently found in patients with hereditary breast or ovarian cancer and present a serious problem for clinical geneticists. This article describes the generation, validation, and application of a reliable high-throughput assay for the functional classification of BRCA1 sequence variants of uncertain significance. Cancer Discov; 3(10); 1142–55. ©2013 AACR.</jats:p><jats:p>This article is highlighted in the In This Issue feature, p. 1083</jats:p>
収録刊行物
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- Cancer Discovery
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Cancer Discovery 3 (10), 1142-1155, 2013-10-01
American Association for Cancer Research (AACR)