Boeri Laura, Radi Orietta, Canzonieri Cecilia, Buscarini Elisabetta, Scatigno Agnese, Minelli Antonella, Ornati Federica, Pagella Fabio, Danesino Cesare, Olivieri Carla
Department of Molecular Medicine, University of Pavia, Crema, Italy.
Mol Syndromol. 2013 Mar;4(3):119-24. doi: 10.1159/000347029. Epub 2013 Feb 28.
Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant vascular dysplasia. Mutations in either ENG or ACVRL1 account for around 85% of cases, and 10% are large deletions and duplications. Here we present a large novel deletion in ACVRL1 gene and its molecular characterization in a 3 generation Italian family. We employed short tandem repeats (STRs) analysis, direct sequencing, multiplex ligation-dependant probe amplification (MLPA) analysis, and 'deletion-specific' PCR methods. STRs Analysis at ENG and ACVRL1 loci suggested a positive linkage for ACVRL1. Direct sequencing of this gene did not identify any mutations, while MLPA identified a large deletion. These results were confirmed and exactly characterized with a 'deletion-specific' PCR: the deletion size is 4,594 bp and breakpoints in exon 3 and intron 8 show the presence of short direct repeats of 7 bp [GCCCCAC]. We hypothesize, as causative molecular mechanism, the replication slippage model. Understanding the fine mechanisms associated with genomic rearrangements may indicate the nonrandomness of these events, highlighting hot spots regions. The complete concordance among MLPA, STRs analysis and 'deletion-specific PCR' supports the usefulness of MLPA in HHT molecular analysis.
遗传性出血性毛细血管扩张症(HHT)是一种常染色体显性血管发育异常疾病。ENG或ACVRL1基因的突变约占病例的85%,10%为大片段缺失和重复。本文报道了一个意大利三代家系中ACVRL1基因的一个新的大片段缺失及其分子特征。我们采用了短串联重复序列(STR)分析、直接测序、多重连接依赖探针扩增(MLPA)分析和“缺失特异性”PCR方法。ENG和ACVRL1位点的STR分析表明ACVRL1存在阳性连锁。该基因的直接测序未发现任何突变,而MLPA检测到一个大片段缺失。这些结果通过“缺失特异性”PCR得到证实并精确表征:缺失大小为4594 bp,外显子3和内含子8中的断点显示存在7 bp的短直接重复序列[GCCCCAC]。我们推测复制滑动模型是其致病分子机制。了解与基因组重排相关的精细机制可能表明这些事件的非随机性,突出热点区域。MLPA、STR分析和“缺失特异性PCR”之间的完全一致性支持了MLPA在HHT分子分析中的实用性。