云南汉族系统性红斑狼疮患者外显子组高通量测序研究

发布时间：2018-05-31 02:12

  本文选题：系统性红斑狼疮 + 基因易感性　； 参考：《昆明医科大学》2017年硕士论文

【摘要】：[目的]系统性红斑狼疮(systemic lupus erythematosus,SLE)的发生发展是遗传、表观遗传、环境等因素共同作用的结果。本研究旨在采用人外显子组高通量测序技术探索云南省汉族人群SLE的遗传因素,筛选云南汉族人群SLE的候选易感基因/变异位点。[方法]随机选取了 2015年10月至2016年6月至我科住院治疗的云南汉族SLE患者作为病例组,以同期云南汉族健康志愿者作为对照组。收集两组人群的基本资料,并采集他们的外周血提取基因组DNA;严格控制DNA质量,按照illumina高通量测序平台操作规范对样本DNA进行人外显子组测序。对获得的测序数据进行比对、质控、过滤、注释、生物信息学分析、易感位点搜寻等分析,以获得SLE易感基因和风险变异信息。[结果]最终纳入汉族SLE患者24人,汉族健康志愿者23人,两组人群年龄、性别均无统计学差异(P0.05)。样本DNA测序平均深度91X,测序质量较高;结果共发现了错义突变(missense)268个(43.79%),框移突变(frameshift deletion)2个(0.33%),剪切子(splicing)区变异1个;发现了 18个外显子区的少见/罕见变异;同时,识别出了 16个SLE变异率明显升高的InDel位点。GO富集分析和kegg pathway分析显示,这些变异位点与淋巴细胞的调控与迁移、机体氧化应激、色氨酸代谢、叶酸代谢、雌激素代谢、唾液分泌等显著相关(P0.01)。综合考虑多方面信息(①最显著差异变异位点(P0.001,OR1),②少见、罕见变异中的致病高风险变异,③发生了框移突变的InDel变异位点),筛选出了 8个变异基因作为候选易感基因。[结论]本研究发现:1)筛选出了8个新的SLE基因多态性位点:rs143467845(SLC4A5,c.A2306G,p.Y769C)、rs74740082(ERC1,c.G1004A,p.R335Q)、rs60797311(KIAA1549,c.G5186A:p.R1729K)、rs117059276(PIGG,c.C1751A:p.S584Y)、rs2228552(COL16A1,c.C185A,p.T62K)、rs1309282(WDR92,c.A721G,p.M241V)等及InDel变异rs138204694(CCDC54,c.647__651del,p.Q216fs)和rs3217319(ZNF480,c.5_6del,p.L2fs),其中5个为新发基因变异(分别是ERC1、PIGG、WDR92、CCDC54和ZNF480),3个基因已被报道过。该8个基因可能在SLE的发生发展中充当着重要的角色,值得进一步扩大样本量验证和进行机制研究。2)虽然GWAS研究目前比较盛行,但基于全外显子组高通量测序(Whole Exome Sequencing,WES)的关联研究具有测序深度深,准确性高,价格相对便宜的特点及优势。也为今后开展更多免疫病遗传学研究打下基础;同时本研究也是第一项基于中国云南省汉族SLE患者的高通量测序研究。
[Abstract]:[objective] the occurrence and development of systemic lupus erythematosus (lupus) is the result of genetic, epigenetic and environmental factors. The aim of this study was to explore the genetic factors of SLE in Yunnan Han population by high throughput sequencing of human exon group and to screen candidate susceptibility genes / mutation sites for SLE in Yunnan Han population. [methods] from October 2015 to June 2016, Yunnan Han nationality patients with SLE were randomly selected as the case group and the healthy volunteers of Yunnan Han nationality as the control group. The basic data of two groups of people were collected and their peripheral blood was collected to extract genomic DNA. The quality of DNA was strictly controlled and the human exon group was sequenced according to the operating specification of illumina high-throughput sequencing platform. The sequence data were compared, quality control, filtering, annotation, bioinformatics analysis, susceptibility site search and so on, to obtain SLE susceptibility gene and risk variation information. [results] there were 24 SLE patients in Han nationality and 23 healthy volunteers in Han nationality. There was no significant difference in age and sex between the two groups. The average depth of DNA sequencing was 91X, and the quality of sequencing was high. A total of 268 missense mutations were found, including 43.79, frameshift deletiontion2, splicing and splicing, respectively, and a rare / rare mutation of 18 exons was found. Sixteen InDel sites with significantly increased SLE mutation rate were identified. Go enrichment analysis and kegg pathway analysis showed that these sites were related to the regulation and migration of lymphocytes, oxidative stress, tryptophan metabolism, folic acid metabolism and estrogen metabolism. Saliva secretion was significantly related to P0.01. Taking into account the most significant variation of information in many aspects, P0.001 and OR1 were rare. The InDel mutation site with frame shift mutation occurred in the high risk mutation of rare mutation, and 8 mutation genes were screened as candidate susceptible genes. [缁撹�篯鏈�鐮旂┒鍙戠帲�

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