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Performant Mutation Identification Using Targeted Next‐Generation Sequencing of 14 Thoracic Aortic Aneurysm Genes
Author(s) -
Proost Dorien,
Vandeweyer Geert,
Meester Josephina A.N.,
Salemink Simone,
Kempers Marlies,
Ingram Christie,
Peeters Nils,
Saenen Johan,
Vrints Christiaan,
Lacro Ronald V.,
Roden Dan,
Wuyts Wim,
Dietz Harry C.,
Mortier Geert,
Loeys Bart L.,
Laer Lut
Publication year - 2015
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.22802
Subject(s) - sanger sequencing , biology , thoracic aortic aneurysm , gene , genetics , candidate gene , multiplex ligation dependent probe amplification , dna sequencing , mutation , genetic diagnosis , genetic testing , aneurysm , aortic aneurysm , bioinformatics , computational biology , exon , medicine , surgery
At least 14 causative genes have been identified for both syndromic and nonsyndromic forms of thoracic aortic aneurysm/dissection (TAA), an important cause of death in the industrialized world. Molecular confirmation of the diagnosis is increasingly important for gene‐tailored patient management but consecutive, conventional molecular TAA gene screening is expensive and labor‐intensive. To circumvent these problems, we developed a TAA gene panel for next‐generation sequencing of 14 TAA genes. After validation, we applied the assay to 100 Marfan patients. We identified 90 FBN1 mutations, 44 of which were novel. In addition, Multiplex ligation‐dependent probe amplification identified large deletions in six of the remaining samples, whereas false‐negative results were excluded by Sanger sequencing of FBN1 , TGFBR1 , and TGFBR2 in the last four samples. Subsequently, we screened 55 syndromic and nonsyndromic TAA patients. We identified causal mutations in 15 patients (27%), one in each of the six following genes: ACTA2, COL3A1, TGFBR1, MYLK, SMAD3, SLC2A10 (homozygous), two in NOTCH1 , and seven in FBN1 . We conclude that our approach for TAA genetic testing overcomes the intrinsic hurdles of consecutive Sanger sequencing of all candidate genes and provides a powerful tool for the elaboration of clinical phenotypes assigned to different genes.