基于RNA测序的SMN1基因缺失型脊髓性肌肉萎缩症的可变剪接差异性分析  

Analysis of the difference of alternative splicing in SMN1 gene deletion type spinal muscular atrophy based on RNA sequencing

作　　者：林炎鸿 张梦雅 曾健 LIN Yanhong;ZHANG Mengya;ZENG Jian(Laboratory of Basic Medicine,Dongfang Hospital(900TH Hospital of Joint Logistics Support Force),Xiamen University/Fujian Provincial Key Laboratory of Transplant Biology,Fuzhou,Fujian 350025,China;Laboratory of Basic Medicine,Fuzong Clinical Medical College of Fujian Medical University(900TH Hospital of Joint Logistics Support Force),Fuzhou,Fujian 350025,China)

机构地区：[1]厦门大学附属东方医院(联勤保障部队第九〇〇医院)基础医学实验室/福建省移植生物学重点实验室,福建福州350025 [2]福建医科大学福总临床医学院(联勤保障部队第九〇〇医院)基础医学实验室,福建福州350025

出　　处：《国际检验医学杂志》2021年第23期2861-2865,共5页International Journal of Laboratory Medicine

基　　金：福建省自然科学基金面上项目(2018J01230、2018J01341)。

摘　　要：目的通过分析运动神经元生存基因1(SMN1)基因纯合性缺失型儿童进行性脊髓性肌肉萎缩症(SMA)患者和SMN1基因拷贝数正常对照外周血淋巴细胞表达谱,研究SMN1基因缺失对可变剪接的影响。方法利用转录组测序对患者组与正常对照组进行表达谱测序,用rMATS软件对RNA-seq数据进行可变剪接事件差异分析,筛选出患者组与正常对照组中差异的外显子跳跃和内含子保留事件,通过在线工具NovoMagic v3.0对这些差异可变剪接体进行基因功能和代谢通路富集分析,实现个性化分析和作图,同时用NCBI数据库对这些基因进行注释。结果(1)SMN1基因纯合性缺失可导致外周血淋巴细胞mRNA可变剪接及其表达量发生变化;(2)外显子跳跃差异基因主要影响核糖核蛋白组装、胞内转运、翻译、乙酰化修饰、线粒体能量代谢及泛素化调控的降解通路;(3)内含子保留差异基因除参与泛素化调控、内吞、翻译、线粒体能量代谢外,同时还调节细胞骨架和代谢酶活性相关乙酰化修饰。结论SMN1基因纯合性缺失能够改变部分基因的可变剪接,进而影响相关蛋白的合成、组装和降解,最终导致蛋白质稳态失衡,为SMA的发病机制提供新的线索。Objective The effect of motor neuron survival gene 1(SMN1)deletion on alternative splicing was studied by analyzing the expression profile of peripheral blood lymphocytes in the homozygous deletion children with progressive spinal muscular atrophy(SMA)and normal SMN1 gene copy number.Methods Used transcriptome sequencing to sequence the expression profile of the patient group and the normal control group.Used rMATS software to analyze the difference of alternative splicing events on RNA-seq data,and screen out the different exon skipping and intron retention events between the patient group and the normal control group.Used the online tool NovoMagic v3.0 to perform gene function and metabolic pathway enrichment analysis of these differential alternative spliceosomes to achieve personalized analysis and mapping.At the same time,the NCBI database was used to annotate these genes.Results(1)Homozygous deletion of SMN1 gene could lead to peripheral blood lymphocyte mRNA alternative splicing and its expression changed;(2)Exon skipping differential genes mainly affected ribonucleoprotein assembly,intracellular transport,translation,acetylation modification,mitochondrial energy metabolism and ubiquitin-regulated degradation pathway.(3)Differential genes with intron retention not only participated in ubiquitin regulation,endocytosis,translation and mitochondrial metabolism,but also regulated acetylation modification related to cytoskeleton and metabolic enzyme activity.Conclusion SMN1 gene homozygous deletion could change the alternative splicing of some genes,and then affect synthesis,assembly and degradation of related proteins,resulting in protein homeostasis imbalance.This provides a new clue to the pathogenesis of SMA.

关 键 词：运动神经元生存基因1纯合性缺失 外周血 RNA测序 可变剪接 内含子保留 外显子跳跃 

分 类 号：R446.1[医药卫生—诊断学]