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, Van Laer Lut
Department of Medical Genetics, Faculty of Medicine and Health Sciences, University of Antwerp and Antwerp University Hospital, Antwerp, Belgium.
Department of Genetics, Radboud University Medical Center, Nijmegen, The Netherlands.
Hum Mutat. 2015 Aug;36(8):808-14. doi: 10.1002/humu.22802. Epub 2015 Jun 13.
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.
对于综合征型和非综合征型胸主动脉瘤/夹层（TAA），已鉴定出至少14个致病基因，TAA是工业化国家重要的死亡原因。分子诊断对于基因定制的患者管理越来越重要，但连续的传统分子TAA基因筛查既昂贵又费力。为了规避这些问题，我们开发了一个用于对14个TAA基因进行下一代测序的TAA基因检测板。验证后，我们将该检测方法应用于100例马凡综合征患者。我们鉴定出90个FBN1突变，其中44个是新突变。此外，多重连接依赖探针扩增在其余六个样本中鉴定出大片段缺失，而通过对最后四个样本中的FBN1、TGFBR1和TGFBR2进行桑格测序排除了假阴性结果。随后，我们对55例综合征型和非综合征型TAA患者进行了筛查。我们在15例患者（27%）中鉴定出致病突变，分别在以下六个基因中的一个：ACTA2、COL3A1、TGFBR1、MYLK、SMAD3、SLC2A10（纯合子），在NOTCH1中有两个，在FBN1中有七个。我们得出结论，我们的TAA基因检测方法克服了对所有候选基因进行连续桑格测序的固有障碍，并为阐述不同基因所对应的临床表型提供了一个强大的工具。
