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| NF1基因新发剪接变异c.3113+5G>T致外显子跳跃的分子机制研究 |
| Research on the de novo heterozygous variant NF1 c.3113+5G>T causing aberrant splicing |
| 投稿时间：2025-08-17 修订日期：2025-11-30 |
| DOI： |
| 中文关键词: NF1 神经纤维瘤病I型 剪接位点变异 外显子跳跃 家系研究 |
| 英文关键词:NF1 Neurofibromatosis type I Splice-site mutation Exon skipping Family study |
| 基金项目:金华市科技计划项目公益类(2022-4-187) |
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| 中文摘要: |
| 目的:神经纤维瘤病I型(Neurofibromatosis type 1,NF1)是一种由NF1基因变异引起的常染色体显性遗传病。NF1基因编码神经纤维瘤蛋白,其功能失活导致Ras信号通路失调,进而引发多系统病变。本研究旨在鉴定一个NF1家系的新发致病变异并阐明其分子机制。方法:收集一个具有典型NF1临床表现(多发牛奶咖啡斑、皮肤神经纤维瘤、身材矮小)家系成员的外周血样本。对先证者进行全外显子测序(WES),利用Sanger测序对家系成员进行变异验证。通过反转录-聚合酶链式反应(RT-PCR)结合琼脂糖凝胶电泳分析变异对mRNA剪接的影响。结果:在先证者及患病家系成员中检出NF1基因内含子23的剪接变异NM_001042492.3:c.3113+5G>T,该变异未见于HGMD(专业版)、ClinVar和gnomAD数据库,属新发变异。RT-PCR及测序显示该变异可导致外显子23跳跃,造成41个氨基酸的缺失(p.Tyr998_Arg1038del),可能影响神经纤维瘤蛋白的功能。根据ACMG指南,评估该变异为“可能致病”。结论:本研究首次报道了NF1基因内含子区杂合剪接位点变异c.3113+5G>T,并通过功能实验证实其导致外显子23跳跃。该发现拓展了NF1基因的变异谱,为NF1的精准基因诊断、遗传咨询及发病机制研究提供了新的分子依据。 |
| 英文摘要: |
| Objective:Neurofibromatosis type 1(NF1)is an autosomal dominant disorder caused by pathogenic variants in the NF1 gene.The NF1 gene encodes neurofibromin,whose functional inactivation disrupts the Ras signaling pathway,leading to multisystem manifestations.This study aimed to identify a novel pathogenic variant in an NF1 family and elucidate its molecular mechanism.Methods:Peripheral blood samples were collected from members of an NF1 family presenting with classic clinical features, including multiple café-au-lait macules, cutaneous neurofibromas, and short stature.Whole-exome sequencing(WES)was performed on the proband,and Sanger sequencing was used to validate the variant in family members.RNA analysis was conducted using reverse transcription-polymerase chain reaction(RT-PCR)and agarose gel electrophoresis to evaluate the effect of the variant on mRNA splicing.Results:A novel heterozygous splice-site variant NF1(NM_001042492.3: c.3113+5G>T)was identified in the proband and affected family members.This variant was absent in public databases,including HGMD(Professional),ClinVar,and gnomAD,and represents a de novo event.RT-PCR and sequencing demonstrated that the variant causes skipping of exon 23,resulting in an in-frame deletion of 41 amino acids(p.Tyr998_Arg1038del),which likely disrupts neurofibromin function. Based on the ACMG/AMP guidelines, the variant was classified as“likely pathogenic”.Conclusion:his study is the first to report the splice-site variant c.3113+5G>T in the NF1 gene and demonstrates through functional assays that it leads to exon 23 skipping.These findings expand the mutational spectrum of the NF1 gene and provide novel molecular insights for precise genetic diagnosis,genetic counseling,and mechanism-based research into NF1. |
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