【作者】 王娜；

【导师】 张福仁；

【作者基本信息】 济南大学 ， 皮肤病与性病学， 2013， 硕士

【摘要】 研究背景：泛发性色素异常症(dyschromatosis universalis hereditaria, DUH)是一种少见的孟德尔遗传性皮肤病，临床上以全身泛发性色素异常伴多系统损害为特征。该病具有高度的遗传异质性，2003年贺林等首次将显性遗传DUH的致病基因定位在6q24.2～q25.2区域，隐性遗传DUH的致病基因定位在12q21～q23区域，其致病基因尚未分离，目前课题组在一个泛发性色素异常症家系中应用全基因组连锁分析，已经将共分离区域定位在2号染色体区间内，这就更提示该病具有很强的遗传异质性。实验目的：①采用全基因组连锁分析及外显子测序相结合的方式，分离DUH的致病基因。②通过表达及其亚细胞定位结合模式、生物斑马鱼体内实验检测致病变异在疾病发生发展过程中的作用，阐述疾病的发病机理。实验方法：①应用Illumina-660K_Beadchips芯片技术对这12个样本进行全基因组分型，应用Merlin软件进行连锁分析；采用Agilent最新的外显子捕获试剂盒；②进行外显子捕获；信息获得后生物信息分析，筛选候选致病变异；③采用Sanger测序法进一步验证，检测基因突变位点。④利用RT-PCR、免疫组织化学和模式动物斑马鱼等实验技术手段，分析致病基因的表达水平，对其功能进行初步探索。实验结果：连锁分析与外显子测序结果分析找到的13个可能的致病变异比对，经Sanger测序验证后筛选致病基因ABCB6，RT-PCR、免疫组织化学和模式动物等实验表明ABCB6是DUH的致病基因。更多还原

【Abstract】 Background: Dyschromatosis universalis hereditaria (DUH) is a rare monogenic disease,characterized by asymptomatic hyper-and hypo-pigmented macules in variable distributionand patterns, which was initially described by Ichikawa and Hiraga in1933. This disorder hasbeen hypothesized to be an autosomal dominant (OMIM127500) or recessive disorder (OMIM612715) according to their kindred’s clinical manifestations. This disorder displays very stronggenetic heterogeneity, although the precise linkage analysis has mapped DUH locus tochromosome6q24.2-q25.22and12q21-q233, the exact molecular basis of DUH remainsunknown. Our research group have used the genomic linkage analysis to find chromosomesregion of the phenotype segregation related from one of DUH pedigree, which have beenlocated on chromosome2, which indicate the strong genetic heterogeneity of DUH.Objective:①We performed genome-wide linkage and exome sequencing analyses in amultiplex Chinese DUH family to find the pathogenic gene.②Through the expression mode analysis, sub-cellular localization et al in vitro experiment,combined with model organism zebrafish in vivo function analysis to verify thedisease-causing gene, which can lay the foundation for the primary prevention.Method:①we performed linkage analyses, genotyping the12individuals of the Family(Family1) using Illumina Human660W-Quad BeadChip. Multipoint parametric linkageanalysis was performed in Merlin11by using pruned autosomal SNPs and assuming adominant inheritance of disease phenotype.②We performed exome sequencing analysis infour affected individuals in order to identify the pathogenic gene.③To verify the mutation，we performed Sanger sequencing and verify the mutation.④Ultimately through theexpression mode analysis, sub-cellular localization et al in vitro experiment, combined withmodel organism zebrafish in vivo function analysis to verify the disease-causing gene.Results: After Exome sequencing, we acquaire13candidate genes, with the comparison withLinkage analysis, and Sange sequencing identified ABCB6as the pathogenic gene. Thefunctional analysis give the further evidence of the gene is the causal gene of DUH.更多还原