Beyond copy number: A new, rapid, and versatile method for sequencing the entire SMN2 gene in SMA patients.
Beyond copy number: A new, rapid, and versatile method for sequencing the entire SMN2 gene in SMA patients.
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超越拷贝数：一种新的，快速且多才多艺的方法，用于对SMA患者的整个SMN2基因进行测序。
DOI：
10.1002/humu.24200
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发表时间：
2021-06
期刊：
影响因子：
3.9
通讯作者：
Tizzano EF
中科院分区：
文献类型：
--
作者：
Blasco-Pérez L;Paramonov I;Leno J;Bernal S;Alias L;Fuentes-Prior P;Cuscó I;Tizzano EF
脊髓性肌萎缩症（SMA）是由SMN 1基因中的双等位基因缺失或致病性变体引起的。SMN 2是SMN 1的高度同源拷贝，被认为是该疾病的主要表型修饰物。确定SMN 2拷贝数对于建立稳健的基因型-表型相关性和预测疾病进展、对临床试验患者进行分层以及确定符合治疗条件的患者至关重要。不一致的基因型-表型相关性在SMA中并不少见，其中一些是由于基因内SMN 2变体可能影响完整SMN转录物的量，因此影响全长SMN蛋白的量。在目前治疗进展的情况下，检测这些变体对于预测SMA表型至关重要，并且从SMA新生儿筛查和早期诊断的角度开始治疗。在这里，我们提出了一种基于远程聚合酶链反应和下一代测序的SMN 2基因完全测序的新型、经济实惠且通用的方法。通过分析53例缺乏SMN 1的SMA患者的样本，对该方法进行了验证，以表征SMN 2以及SMN 2-SMN 1杂合基因的旁系同源、罕见变异和单核苷酸多态性。该方法鉴定部分缺失，并可适用于确定具有至少一个SMN 1拷贝的患者中的罕见致病性变体。我们开发了一种新的经济实惠的通用方法，通过长PCR和NGS，可以对脊髓性肌萎缩症（SMA）患者的整个SMN 2基因进行测序，检测点变异，拷贝数变异和混合SMN 1-SMN 2基因。对每例患者SMN 2结构的完整表征将提高我们对基因型-表型相关性、新生儿筛查和症状前诊断背景下的表型预测的认识，还可能允许识别新的表型修饰基因变体。
Spinal muscular atrophy (SMA) is caused by bi‐allelic loss or pathogenic variants in the SMN1 gene. SMN2, the highly homologous copy of SMN1, is considered the major phenotypic modifier of the disease. Determination of SMN2 copy number is essential to establish robust genotype–phenotype correlations and predict disease evolution, to stratify patients for clinical trials, as well as to define those eligible for treatment. Discordant genotype–phenotype correlations are not uncommon in SMA, some of which are due to intragenic SMN2 variants that may influence the amount of complete SMN transcripts and, therefore, of full‐length SMN protein. Detection of these variants is crucial to predict SMA phenotypes in the present scenario of therapeutic advances and with the perspective of SMA neonatal screening and early diagnosis to start treatments. Here, we present a novel, affordable, and versatile method for complete sequencing of the SMN2 gene based on long‐range polymerase chain reaction and next‐generation sequencing. The method was validated by analyzing samples from 53 SMA patients who lack SMN1, allowing to characterize paralogous, rare variants, and single‐nucleotide polymorphisms of SMN2 as well as SMN2–SMN1 hybrid genes. The method identifies partial deletions and can be adapted to determine rare pathogenic variants in patients with at least one SMN1 copy. We developed a new affordable and versatile method that, by means of long PCR and NGS, allows sequencing the entire SMN2 genes of spinal muscular atrophy (SMA) patients detecting point variants, copy number variants and hybrid SMN1‐SMN2 genes. This complete characterization of the SMN2 structure in each patient will improve our knowledge of the genotype‐phenotype correlation, the phenotype prediction in the context of newborn screening and pre‐symptomatic diagnosis and also will potentially allow the identification of new phenotype modifier variants.
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影响因子：
64.5
作者：
LEFEBVRE, S;BURGLEN, L;MELKI, J
通讯作者：
MELKI, J
DOI：
10.1038/ejhg.2011.134
发表时间：
2012-01
期刊：
European journal of human genetics : EJHG
影响因子：
--
作者：
通讯作者：
--
影响因子：
1.4
作者：
Alias, Laura;Bernal, Sara;Tizzano, Eduardo F.
通讯作者：
Tizzano, Eduardo F.
DOI：
10.1212/nxg.0000000000000530
发表时间：
2020-12
期刊：
Neurology. Genetics
影响因子：
--
作者：
Cuscó I;Bernal S;Blasco-Pérez L;Calucho M;Alias L;Fuentes-Prior P;Tizzano EF
通讯作者：
Tizzano EF
影响因子：
5.3
作者：
Alias, Laura;Bernal, Sara;Tizzano, Eduardo F.
通讯作者：
Tizzano, Eduardo F.