PT - JOURNAL ARTICLE AU - Massimo Bogliolo AU - Roser Pujol AU - Miriam Aza-Carmona AU - Núria Muñoz-Subirana AU - Benjamin Rodriguez-Santiago AU - José Antonio Casado AU - Paula Rio AU - Christopher Bauser AU - Judith Reina-Castillón AU - Marcos Lopez-Sanchez AU - Lidia Gonzalez-Quereda AU - Pia Gallano AU - Albert Catalá AU - Ana Ruiz-Llobet AU - Isabel Badell AU - Cristina Diaz-Heredia AU - Raquel Hladun AU - Leonort Senent AU - Bienvenida Argiles AU - Juan Miguel Bergua Burgues AU - Fatima Bañez AU - Beatriz Arrizabalaga AU - Ricardo López Almaraz AU - Monica Lopez AU - Ángela Figuera AU - Antonio Molinés AU - Inmaculada Pérez de Soto AU - Inés Hernando AU - Juan Antonio Muñoz AU - Maria del Rosario Marin AU - Judith Balmaña AU - Neda Stjepanovic AU - Estela Carrasco AU - Isabel Cuesta AU - José Miguel Cosuelo AU - Alexandra Regueiro AU - José Moraleda Jimenez AU - Ana Maria Galera-Miñarro AU - Laura Rosiñol AU - Anna Carrió AU - Cristina Beléndez-Bieler AU - Antonio Escudero Soto AU - Elena Cela AU - Gregorio de la Mata AU - Rafael Fernández-Delgado AU - Maria Carmen Garcia-Pardos AU - Raquel Sáez-Villaverde AU - Marta Barragaño AU - Raquel Portugal AU - Francisco Lendinez AU - Ines Hernadez AU - José Manue Vagace AU - Maria Tapia AU - José Nieto AU - Marta Garcia AU - Macarena Gonzalez AU - Cristina Vicho AU - Eva Galvez AU - Alberto Valiente AU - Maria Luisa Antelo AU - Phil Ancliff AU - Francisco Garcia AU - Joaquin Dopazo AU - Julian Sevilla AU - Tobias Paprotka AU - Luis Alberto Pérez-Jurado AU - Juan Bueren AU - Jordi Surralles TI - Optimised molecular genetic diagnostics of Fanconi anaemia by whole exome sequencing and functional studies AID - 10.1136/jmedgenet-2019-106249 DP - 2019 Oct 05 TA - Journal of Medical Genetics PG - jmedgenet-2019-106249 4099 - http://jmg.bmj.com/content/early/2019/10/05/jmedgenet-2019-106249.short 4100 - http://jmg.bmj.com/content/early/2019/10/05/jmedgenet-2019-106249.full AB - Purpose Patients with Fanconi anaemia (FA), a rare DNA repair genetic disease, exhibit chromosome fragility, bone marrow failure, malformations and cancer susceptibility. FA molecular diagnosis is challenging since FA is caused by point mutations and large deletions in 22 genes following three heritability patterns. To optimise FA patients’ characterisation, we developed a simplified but effective methodology based on whole exome sequencing (WES) and functional studies.Methods 68 patients with FA were analysed by commercial WES services. Copy number variations were evaluated by sequencing data analysis with RStudio. To test FANCA missense variants, wt FANCA cDNA was cloned and variants were introduced by site-directed mutagenesis. Vectors were then tested for their ability to complement DNA repair defects of a FANCA-KO human cell line generated by TALEN technologies.Results We identified 93.3% of mutated alleles including large deletions. We determined the pathogenicity of three FANCA missense variants and demonstrated that two FANCA variants reported in mutations databases as ‘affecting functions’ are SNPs. Deep analysis of sequencing data revealed patients’ true mutations, highlighting the importance of functional analysis. In one patient, no pathogenic variant could be identified in any of the 22 known FA genes, and in seven patients, only one deleterious variant could be identified (three patients each with FANCA and FANCD2 and one patient with FANCE mutations)Conclusion WES and proper bioinformatics analysis are sufficient to effectively characterise patients with FA regardless of the rarity of their complementation group, type of mutations, mosaic condition and DNA source.