Imbard Apolline, Pasmant Eric, Sabbagh Audrey, Luscan Armelle, Soares Magali, Goussard Philippe, Blanché Hélène, Laurendeau Ingrid, Ferkal Salah, Vidaud Michel, Pinson Stéphane, Bellanne-Chantelot Christine, Vidaud Dominique, Wolkenstein Pierre, Parfait Béatrice
Service de Biochimie-Hormonologie, Hôpital Robert Debré, Assistance Publique-Hôpitaux de Paris, Paris, France.
1] EA7331, Université Paris Descartes, Sorbonne Paris Cité, Faculté des Sciences Pharmaceutiques et Biologiques, Paris, France [2] Service de Biochimie et de Génétique Moléculaire, Hôpital Cochin, Assistance Publique-Hôpitaux de Paris, Paris, France.
J Hum Genet. 2015 Apr;60(4):221-4. doi: 10.1038/jhg.2015.6. Epub 2015 Jan 29.
Neurofibromatosis type 1 (NF1) is caused by dominant loss-of-function mutations of the tumor suppressor NF1 containing 57 constitutive coding exons. A huge number of different pathogenic NF1 alterations has been reported. The aim of the present study was to evaluate the usefulness of a multiplex ligation-dependent probe amplification (MLPA) approach in NF1 patients to detect single and multi-exon NF1 gene copy number variations. A genotype-phenotype correlation was then performed in NF1 patients carrying these types of genetic alterations. Among 565 NF1 index cases from the French NF1 cohort, single and multi-exon deletions/duplications screening identified NF1 partial deletions/duplications in 22 patients (~4%) using MLPA analysis. Eight single exon deletions, 11 multiple exons deletions, 1 complex rearrangement and 2 duplications were identified. All results were confirmed using a custom array-CGH. MLPA and custom array-CGH allowed the identification of rearrangements that were missed by cDNA/DNA sequencing or microsatellite analysis. We then performed a targeted next-generation sequencing of NF1 that allowed confirmation of all 22 rearrangements. No clear genotype-phenotype correlations were found for the most clinically significant disease features of NF1 in patients with single and multi-exons NF1 gene copy number changes.
1型神经纤维瘤病（NF1）由肿瘤抑制基因NF1的显性功能丧失突变引起，NF1含有57个组成型编码外显子。已报道了大量不同的致病性NF1改变。本研究的目的是评估多重连接依赖探针扩增（MLPA）方法在NF1患者中检测单外显子和多外显子NF1基因拷贝数变异的实用性。然后对携带这些类型基因改变的NF1患者进行基因型-表型相关性分析。在法国NF1队列的565例NF1索引病例中，使用MLPA分析对单外显子和多外显子缺失/重复进行筛查，在22例患者（约4%）中鉴定出NF1部分缺失/重复。鉴定出8个单外显子缺失、11个多外显子缺失、1个复杂重排和2个重复。所有结果均使用定制的阵列比较基因组杂交（array-CGH）进行了确认。MLPA和定制阵列CGH能够鉴定出cDNA/DNA测序或微卫星分析遗漏的重排。然后我们对NF1进行了靶向二代测序，从而确认了所有22种重排。在单外显子和多外显子NF1基因拷贝数改变的患者中，未发现与NF1最具临床意义的疾病特征有明确的基因型-表型相关性。
