原发性干燥综合征易感基因与子痫前期候选基因的研究

发布时间：2018-04-15 19:26

本文选题：原发性干燥综合征 + 全外显子测序　；参考：《北京协和医学院》2017年硕士论文

【摘要】：第一部分全外显子测序探究原发性干燥综合征易感基因的研究研究背景:干燥综合征(Sjogren's syndrome,SS)是一种常见的自身免疫性疾病,临床上典型表现为淋巴细胞浸润外分泌腺,尤其是唾液腺及泪腺,造成腺体破坏,分泌功能受损。患者除表现为口眼干燥外,随病情进展常累及全身各个系统器官。干燥综合征如果单独发生,即为原发性干燥综合征(Primary Sjogren's syndrome,pSS),若伴随其它免疫性疾病如系统性红斑狼疮、风湿性关节炎等发生时,则为继发性干燥综合征(Secondary Sjogren's syndrome,sSS)。原发性干燥综合征为常见病,在系统性自身免疫疾病中发病率仅次于风湿性关节炎。目前,干燥综合征的发病机制尚不明确,疾病的发生发展过程涉及环境因素与遗传因素的共同作用。pSS易感基因的研究是当前对该病的研究热点之一,以往的研究多采用候选基因法,近年有两篇分别在白种人和中国汉族人群中开展的GWAS,发现了新的与pSS相关的基因。研究目的:以往的研究发现的位点多为常见变异且多数位于基因的内含子或基因间区域,对其功能的研究较为欠缺,本研究拟采用全外显子测序技术(Whole exome sequence,WES)研究pSS,尝试寻找与中国汉族人群pSS相关的外显子区变异。研究方法:收集诊断明确的pSS患者共91例,健康对照152例,采集患者及对照的基本信息及外周血样本,并提取基因组DNA,保存备用;对11例pSS患者及6例健康对照采用Ion Proton测序平台进行测序;得到的序列信息经TS系统完成比对和变异读出等常规分析,IR系统内置的分析模块完成变异位点的生物学功能注释及报告生成等重要步骤。对变异位点的注释信息包括基因名称、染色体位置、氨基酸变化、GO注释、人群频率及SIFT、Polyphen预测结果等,综合考虑以上信息筛选变异位点并进行验证,以探索可能与pSS相关变异位点及基因。研究结果:1.通过NCBI-Gene数据库检索到可能与pSS相关的基因共171个,从WES结果中整理出位于这些基因区域内的变异,经筛选后对其中验证无误的5个变异位点,包括E7S1 c.*3297GA、DARCc.205CT、DARCc.131GA、KIR2DL1 g.55295367GA和KIR2DL3 c.686CT在80例散发病例及146例健康对照中直接测序,这五个位点的分型结果显示等位基因频率及基因型分布差异无统计学意义(P0.05).2、通过对WES数据的分析,筛选到25个罕见变异位点(MAF0.01),这些位点在6例对照中均未发现。对这25个位点所在基因进行功能富集分析发现IFIH1、CREBBP,OAS1 富集到 Influenza A(P0.05)、Herpes simplex infection(P0.05)及 Hepatitis B(P0.05)信号通路。结论:1、在中国汉族人群中位点ETS1 c.*3297GA、DARC c.205CT、DARC c.131GA、KIR2DL1g.55295367GA和KIR2D 3 c.686CT 可能与 pSS 不相关。2、通过对本研究中发现的25个罕见变异所在基因进行功能分析发现IFIH1、CREBBP、OAS1可能是pSS易感基因,但仍需验证进行确认。第二部分中国汉族妇女子痫前期的候选基因关联研究目的:子痫前期(Preeclampsia,PE)是发生于妊娠期20周以后,以新发高血压和蛋白尿为发病特征的妊娠期高血压疾病。PE被认为是一种胎盘源性疾病,患者在胎盘娩出后常很快缓解或可自愈。发病机制目前尚不清楚,其病理过程主要涉及遗传因素、免疫因素、胎盘缺血、内皮细胞损伤及氧化应激。本研究以181例PE患者(病例组)及203例健康孕妇(对照组)作为研究对象,对2q14.2区域的三个 SNP 位点 rs12711941、rs7576192、rs7579169,分别位于基因ANG-1、SOD2、SOD3 的 SNP 位点 rs2507800、rs4880、rs1799895及TIP3 基因的 9 个tagSNP位点与中国汉族妇女PE的相关性进行了分析,以期发现与中国汉族妇女PE相关的位点及基因。方法:抽取181例PE患者及203例健康孕妇的外周血,提取基因组DNA,采用飞行时间质谱法对以上15个位点进行基因分型。采用SPSS12.0对分型结果进行统计分析,计算每个位点的等位基因频率及基因型频率在PE患者及健康孕妇中是否存在差异。结果:rs7579169位点等位基因频率在两组间有差异(P=0.02,OR=1.44,95%CI 1.05-1.98),在显隐性及加性模型下分别对该位点进行分析,结果显示在显性模型下PE组与对照组存在差异(P=0.040,OR=2.278)。SNP位点rs7576192、rs12711941等位基因频率及基因型频率在病例组及对照组间均无统计学差异(P=0.12,P=0.37),分别在显性、隐性及加性模型下对这两个位点进行分析,两组间均无统计学差异。对位点rs12711941、rs7576192和rs7579169组成的单体型进行分析,结果显示单体型GGC和GGT在PE患者及对照组间均存在差异(P=0.030,P=0.046)。多态位点 rs2507800、rs4880、rs1799895 等位基因频率和基因型频率在两组间无统计学差异。对位点rs2507800、rs4880分别在显隐性模型下进行分析,仍无统计学差异。对TIMP3基因9个tagSNP的分析显示,这些位点的等位基因频率在两组间无差异,但多次分娩(3次)的PE患者亚组与对照组相比,rs135025位点基因型分布在加性模型及显性模型下差异有统计学意义(P=0.018,P=0.008)。位点rs80272基因型分布在加性模型及显性模型下,重度子痫前期组与对照组相比有统计学差异(P=0.014,P=0.041)。对这9个tagSNP(rs135025、rs135029、rs137487、rs241890、rs242078、rs5754312、rs715572、rs80272和rs9609643)位点进行单体型分析,发现单体型ACGTAAGCG在PE组与对照组间差异有统计学意义(P=0.035)。结论:rs7579169 与 PE 相关,rs7576192 及 rs12711941 与 PE 不相关;rs12711941、rs7576192和rs7579169组成的单体型GGC和GGT与PE相关。基因TIMP3的9 个 tagSNP 位点均与 PE 不相关,但由 rs135025、rs135029、rs137487、rs241890、rs242078、rs5754312、rs715572、rs80272 和 rs9609643 组成的单体型 ACGTAAG CG 与 PE 相关。基因 ANG-1、SOD2和SOD3上的 SNP 位点 rs2507800、rs4880和rs1799895均与PE不相关。
[Abstract]:Sjogren ' s syndrome ( SS ) is a common autoimmune disease . The results of this study were as follows : 1 . A total of 171 patients with pSS related to pSS were collected from WES . The results were as follows : 1 . The results were as follows : 1 . The results were as follows : 1 . The results showed that there were 171 patients with pSS related to pSS . The results were as follows : 1 . The results showed that there were no significant differences in the gene name , chromosome position , amino acid change , GO annotation , population frequency and the results of the study . In the second part , the genetic factors , immune factors , placenta ischemia , endothelial cell injury and oxidative stress were identified . Results : The allele frequency of rs7579169 locus was different between the two groups ( P = 0.02 , OR = 1.44 , 95 % CI 1.05 - 1.98 ) . The results showed that there was a difference between the PE group and the control group ( P = 0 . 040 , OR = 2.278 ) . There was no statistical difference between the two groups ( P = 0.12 , P = 0.37 ) . There was no statistical difference between the two groups . The results showed that the haplotype GGC and GGT were different in PE patients and control groups ( P = 0.030 , P = 0.046 ) . The frequencies of rs2507800 , rs4880 , rs1799895 , rs4880 and rs1799895 were not significantly different between the two groups . The genotype distribution of rs80272 genotype was statistically different from the control group ( P = 0.014 , P = 0.041 ) . Conclusion : rs7579169 is related to PE , rs7576192 and rs1211941 are not related to PE . Conclusion : The single type of GGC and GGT are not related to PE . rs135025 , rs7576192 and rs7579169 are not related to PE . The rs135025 , rs135029 , rs137487 , rs24915572 , rs242078 , rs5754312 , rs715572 , rs80272 and rs96,4343 are not related to PE . The SNP loci rs2507800 , rs4880 and rs1799895 on the gene ANG - 1 , SOD2 and SOD3 are not related to PE .

【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R593.2;R714.244

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