Mondéjar Rufino, Solano Francisca, Rubio Rocío, Delgado Mercedes, Pérez-Sempere Angel, González-Meneses Antonio, Vendrell Teresa, Izquierdo Guillermo, Martinez-Mir Amalia, Lucas Miguel
Servicio de Biología Molecular, Hospital Universitario Virgen Macarena, Facultad de Medicina, Sevilla, Spain.
Servicio de Neurología, Hospital Universitario de Alicante, Alicante, Spain.
PLoS One. 2014 Jan 23;9(1):e86286. doi: 10.1371/journal.pone.0086286. eCollection 2014.
To study the molecular genetic and clinical features of cerebral cavernous malformations (CCM) in a cohort of Spanish patients.
We analyzed the CCM1, CCM2, and CCM3 genes by MLPA and direct sequencing of exons and intronic boundaries in 94 familial forms and 41 sporadic cases of CCM patients of Spanish extraction. When available, RNA studies were performed seeking for alternative or cryptic splicing.
A total of 26 pathogenic mutations, 22 of which predict truncated proteins, were identified in 29 familial forms and in three sporadic cases. The repertoire includes six novel non-sense and frameshift mutations in CCM1 and CCM3. We also found four missense mutations, one of them located at the third NPXY motif of CCM1 and another one that leads to cryptic splicing of CCM1 exon 6. We found four genomic deletions with the loss of the whole CCM2 gene in one patient and a partial loss of CCM1and CCM2 genes in three other patients. Four families had mutations in CCM3. The results include a high frequency of intronic variants, although most of them localize out of consensus splicing sequences. The main symptoms associated to clinical debut consisted of cerebral haemorrhage, migraines and epileptic seizures. The rare co-occurrence of CCM with Noonan and Chiari syndromes and delayed menarche is reported.
Analysis of CCM genes by sequencing and MLPA has detected mutations in almost 35% of a Spanish cohort (36% of familial cases and 10% of sporadic patients). The results include 13 new mutations of CCM genes and the main clinical symptoms that deserves consideration in molecular diagnosis and genetic counselling of cerebral cavernous malformations.
研究一组西班牙患者脑海绵状血管畸形（CCM）的分子遗传学及临床特征。
我们采用多重连接探针扩增技术（MLPA）以及对94例家族性病例和41例西班牙裔散发CCM患者的外显子及内含子边界进行直接测序，分析CCM1、CCM2和CCM3基因。如有可能，进行RNA研究以寻找替代或隐匿性剪接。
在29例家族性病例和3例散发病例中，共鉴定出26个致病突变，其中22个预测会产生截短蛋白。该突变谱包括CCM1和CCM3中的6个新的无义突变和移码突变。我们还发现了4个错义突变，其中1个位于CCM1的第三个NPXY基序，另1个导致CCM1外显子6的隐匿性剪接。我们发现1例患者存在整个CCM2基因缺失的4个基因组缺失，另外3例患者存在CCM1和CCM2基因部分缺失。4个家族的CCM3存在突变。结果显示内含子变异频率较高，尽管其中大多数位于共有剪接序列之外。临床首发的主要症状包括脑出血、偏头痛和癫痫发作。报告了CCM与努南综合征和Chiari综合征罕见的共现情况以及初潮延迟。
通过测序和MLPA分析CCM基因，在一组西班牙患者中检测到近35%的突变（家族性病例中为36%，散发性患者中为10%）。研究结果包括CCM基因的13个新突变以及脑海绵状血管畸形分子诊断和遗传咨询中值得考虑的主要临床症状。
