自身免疫相关基因ZMIZ1及其调控SNPrs1408944在白癜风发病中的机制研究

ZMIZ1基因既往在多个欧洲种群的研究中被证实为多种自身免疫性疾病相关的易感基因，2014年课题申请人利用大规模基因分型实验成功证明ZMIZ1基因在中国人群中与白癜风相关联，同时利用多种生物信息学方法预测其中的单核苷酸多态性位点rs1408944可以通过结合多个转录调控因子调节ZMIZ1基因的表达，但此种机制尚未通过分子生物学方法予以证实。本项目拟利用斑马鱼模式动物、酪氨酸酶活力和黑色素蛋白含量测定、通路基因共表达等多种分子生物学方法对ZMIZ1基因在自身免疫途径及黑素合成中的作用进行研究；同时利用荧光素酶报告基团实验、ChIP-qPCR（染色质免疫共沉淀－定量PCR）、B－淋巴细胞增殖率等实验对rs1408944调控ZMIZ1基因表达的机制进行深入研究，以期从基因组和转录调控水平上揭示ZMIZ1基因在白癜风发病中的作用机制。

ZMIZ1 has been identified as susceptibility gene to be associated with multiple autoimmune disorders by genome-wide association study in European population. In 2014, we have performed a large-scale genotyping analysis as well as bioinformatics study to investigate the effect of ZMIZ1 on vitiligo in Chinese population, which identified rs1408944 to be associated with vitiligo and had a functional effect by creating a transcription factor binding site to multiple transcription factors. However, the functional prediction has not been confirmed by molecular experiments. In this study, we are planning to perform sufficient experiments to investigate the function of ZMIZ1 in the autoimmune as well as the melanin synthesis process, such as gene knock-down zebrafish model, estimation of the tyrosinase activity and melanin quantitive experiment. Further, to investigate the regulatory role of SNP rs1408944 to ZMIZ1 gene, molecular experiments such as luciferase reporter assay, ChiP-qPCR and B-cell proliferation rate will be performed to elucidate the functional role of both ZMIZ1 and its regulatory variant in the pathogenesis of vitiligo.