ASAH1基因的一个新的内含子变异增强了异常剪接,导致脊髓性肌萎缩伴进行性肌阵挛性癫痫。

IF 3.2 3区 医学 Q1 PEDIATRICS
Jinli Bai, Ping Li, Hui Jiao, Yuwei Jin, Hong Wang, Qinglin Jiang, Fang Song, Xiaoyin Peng, Yujin Qu
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引用次数: 0

摘要

背景:脊髓性肌萎缩伴进行性肌阵挛性癫痫(SMA-PME)是一种罕见的常染色体隐性遗传病,由ASAH1基因变异引起。虽然ASAH1编码变异体会导致SMA-PME,但非编码变异体,特别是非规范剪接位点变异体的影响尚不清楚。方法:先证者进行全外显子组测序(WES),并采用Sanger测序法确认核心家族成员中变异的携带者状态。通过互补DNA (cDNA)和迷你基因剪接实验来验证剪接效果。结果:通过WES鉴定出两个杂合ASAH1变异:c.304dupA (p.Thr102Asnfs*14)和c.264 + 11A > G。Sanger测序证实,这些变异在反式中是双亲分离的:c.304dupA遗传自父亲,c.264 + 11A > G遗传自母亲。根据ACMG指南,c.304dupA变异被归类为致病性。然而,第3内含子中的c.264 + 11A > G变异是首次报道,其功能影响尚未完全阐明。互补DNA (cDNA)和迷你基因剪接实验表明,c.264 + 11A >g变异产生了两个转录本。携带c.264 + 11A > G等位基因的ASAH1转录本中约有10%是全长的,而其余转录本缺乏外显子3。外显子跳变是由剪接异常引起的,这可能导致过早终止密码子(PTC, p.Tyr59Ter)。结论:据我们所知,c.264 + 11A >g是该基因中第一个可能的致病非典型剪接位点变异。这通过外显子3跳变驱动SMA-PME的发病机制。我们的研究结果为非典型剪接位点变异的复杂剪接机制提供了新的见解,强调了cDNA分析和迷你基因剪接测定在SMA-PME病例的精确诊断和遗传咨询中的独特作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A novel intronic variant in the ASAH1 gene enhances aberrant splicing, causing spinal muscular atrophy with progressive myoclonic epilepsy.

Background: Spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME) is a rare autosomal recessive disorder caused by ASAH1 gene variants. Although ASAH1 coding variants cause SMA-PME, the impact of noncoding variants, particularly noncanonical splice-site variants, is less clear.

Methods: Whole-exome sequencing (WES) was performed on the proband, and Sanger sequencing was used to confirm the carrier status of the variants in the core family members. Complementary DNA (cDNA) and minigene splicing assays were performed to validate the splicing effects.

Results: Two heterozygous ASAH1 variants were identified through WES: c.304dupA (p.Thr102Asnfs*14) and c.264 + 11A > G. Sanger sequencing confirmed that the variants were bi-parentally segregated in trans: c.304dupA was inherited from the father, and c.264 + 11A > G was inherited from the mother. The c.304dupA variant was classified as pathogenic according to the ACMG guidelines. However, the c.264 + 11A > G variant in intron 3 was reported for the first time, and its functional impact has not yet been fully elucidated. Complementary DNA (cDNA) and minigene splicing assays indicated that the c.264 + 11A > G variant generated two transcripts. Approximately 10% of the ASAH1 transcripts from the allele carrying c.264 + 11A > G were full length, whereas the remaining transcripts lacked exon 3. Exon skipping results from aberrant splicing, which potentially leads to a premature termination codon (PTC, p.Tyr59Ter).

Conclusion: To the best of our knowledge, the c.264 + 11A > G is the first likely pathogenic noncanonical splice-site variant identified in this gene. This drives the pathogenesis of SMA-PME through exon 3 skipping. Our findings provide new insights into the intricate splicing mechanisms of noncanonical splice-site variants, emphasizing the unique role of cDNA analysis and minigene splicing assays in the precise diagnosis and genetic counseling of SMA-PME cases.

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来源期刊
CiteScore
6.10
自引率
13.90%
发文量
192
审稿时长
6-12 weeks
期刊介绍: Italian Journal of Pediatrics is an open access peer-reviewed journal that includes all aspects of pediatric medicine. The journal also covers health service and public health research that addresses primary care issues. The journal provides a high-quality forum for pediatricians and other healthcare professionals to report and discuss up-to-the-minute research and expert reviews in the field of pediatric medicine. The journal will continue to develop the range of articles published to enable this invaluable resource to stay at the forefront of the field. Italian Journal of Pediatrics, which commenced in 1975 as Rivista Italiana di Pediatria, provides a high-quality forum for pediatricians and other healthcare professionals to report and discuss up-to-the-minute research and expert reviews in the field of pediatric medicine. The journal will continue to develop the range of articles published to enable this invaluable resource to stay at the forefront of the field.
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