Identification of deep intronic variants in junctional epidermolysis bullosa using genome sequencing and splicing assays.

IF 4.7 2区医学 Q1 GENETICS & HEREDITY

NPJ Genomic Medicine Pub Date : 2025-02-06 DOI:10.1038/s41525-025-00466-8

Fuying Chen, Ruoqu Wei, Yumeng Wang, Qiaoyu Cao, Jianbo Wang, Chenfei Wang, Dingjin Yao, Zhirong Yao, Cheng Ni, Ming Li

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Abstract

Junctional epidermolysis bullosa (JEB) is characterized by mucocutaneous fragility. We enrolled 69 cases of recessive JEB, with 13.0% of these cases remained genetically undiagnosed following an initial exome sequencing. Among cases carried COL17A1 variants, this proportion can reach 31.6%. We employed genome sequencing to genetically diagnosis these cases. Four deep intronic variants (c.4156+117 G > A, c.2039-104 G > A and c.1267+237dupC in the COL17A1 gene and c.-38 + 2 T > C in the LAMB3 gene) were identified in six cases. The c.4156+117 G > A variant was found in three of the five cases, suggesting it may be a common deep intronic variant in Chinese JEB. Splicing analysis revealed that these variants caused splicing defect by inducing exon skipping, or pseudoexon insertion into the transcript in HaCaT cells, not in HEK293 cells. Our results emphasize the importance of selecting the right cell line for mRNA analysis.

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利用基因组测序和剪接技术鉴定大疱性结性表皮松解症的深层内含子变异。

大疱性结缔组织表皮松解症（JEB）以皮肤粘膜脆性为特征。我们招募了69例隐性JEB病例，其中13.0%的病例在初始外显子组测序后仍未得到遗传诊断。在携带COL17A1变异的病例中，这一比例可达31.6%。我们采用基因组测序对这些病例进行基因诊断。在6例患者中发现了COL17A1基因中的C .4156+117 G > A、C .2039-104 G > A、C .1267+237dupC和LAMB3基因中的C .-38 +2 T > C四个深层内含子变异。5例病例中有3例发现c.4156+117 G > A变异，提示它可能是中国JEB常见的深内含子变异。剪接分析表明，这些变异通过诱导外显子跳变或假外显子插入到HaCaT细胞的转录本中而不是在HEK293细胞中引起剪接缺陷。我们的结果强调了选择正确细胞系进行mRNA分析的重要性。

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来源期刊

NPJ Genomic Medicine Biochemistry, Genetics and Molecular Biology-Molecular Biology

CiteScore

9.40

自引率

1.90%

发文量

审稿时长

17 weeks

期刊介绍： npj Genomic Medicine is an international, peer-reviewed journal dedicated to publishing the most important scientific advances in all aspects of genomics and its application in the practice of medicine. The journal defines genomic medicine as "diagnosis, prognosis, prevention and/or treatment of disease and disorders of the mind and body, using approaches informed or enabled by knowledge of the genome and the molecules it encodes." Relevant and high-impact papers that encompass studies of individuals, families, or populations are considered for publication. An emphasis will include coupling detailed phenotype and genome sequencing information, both enabled by new technologies and informatics, to delineate the underlying aetiology of disease. Clinical recommendations and/or guidelines of how that data should be used in the clinical management of those patients in the study, and others, are also encouraged.