本文選題：系統(tǒng)性紅斑狼瘡 + 基因易感性�。� 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：[目的]系統(tǒng)性紅斑狼瘡(systemic lupus erythematosus,SLE)的發(fā)生發(fā)展是遺傳、表觀遺傳、環(huán)境等因素共同作用的結(jié)果。本研究旨在采用人外顯子組高通量測序技術(shù)探索云南省漢族人群SLE的遺傳因素,篩選云南漢族人群SLE的候選易感基因/變異位點。[方法]隨機(jī)選取了 2015年10月至2016年6月至我科住院治療的云南漢族SLE患者作為病例組,以同期云南漢族健康志愿者作為對照組。收集兩組人群的基本資料,并采集他們的外周血提取基因組DNA;嚴(yán)格控制DNA質(zhì)量,按照illumina高通量測序平臺操作規(guī)范對樣本DNA進(jìn)行人外顯子組測序。對獲得的測序數(shù)據(jù)進(jìn)行比對、質(zhì)控、過濾、注釋、生物信息學(xué)分析、易感位點搜尋等分析,以獲得SLE易感基因和風(fēng)險變異信息。[結(jié)果]最終納入漢族SLE患者24人,漢族健康志愿者23人,兩組人群年齡、性別均無統(tǒng)計學(xué)差異(P0.05)。樣本DNA測序平均深度91X,測序質(zhì)量較高;結(jié)果共發(fā)現(xiàn)了錯義突變(missense)268個(43.79%),框移突變(frameshift deletion)2個(0.33%),剪切子(splicing)區(qū)變異1個;發(fā)現(xiàn)了 18個外顯子區(qū)的少見/罕見變異;同時,識別出了 16個SLE變異率明顯升高的InDel位點。GO富集分析和kegg pathway分析顯示,這些變異位點與淋巴細(xì)胞的調(diào)控與遷移、機(jī)體氧化應(yīng)激、色氨酸代謝、葉酸代謝、雌激素代謝、唾液分泌等顯著相關(guān)(P0.01)。綜合考慮多方面信息(①最顯著差異變異位點(P0.001,OR1),②少見、罕見變異中的致病高風(fēng)險變異,③發(fā)生了框移突變的InDel變異位點),篩選出了 8個變異基因作為候選易感基因。[結(jié)論]本研究發(fā)現(xiàn):1)篩選出了8個新的SLE基因多態(tài)性位點: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變異rs138204694(CCDC54,c.647__651del,p.Q216fs)和rs3217319(ZNF480,c.5_6del,p.L2fs),其中5個為新發(fā)基因變異(分別是ERC1、PIGG、WDR92、CCDC54和ZNF480),3個基因已被報道過。該8個基因可能在SLE的發(fā)生發(fā)展中充當(dāng)著重要的角色,值得進(jìn)一步擴(kuò)大樣本量驗證和進(jìn)行機(jī)制研究。2)雖然GWAS研究目前比較盛行,但基于全外顯子組高通量測序(Whole Exome Sequencing,WES)的關(guān)聯(lián)研究具有測序深度深,準(zhǔn)確性高,價格相對便宜的特點及優(yōu)勢。也為今后開展更多免疫病遺傳學(xué)研究打下基礎(chǔ);同時本研究也是第一項基于中國云南省漢族SLE患者的高通量測序研究。
[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. [緇撹]鏈爺絀跺彂鐜，

本文編號：1957869