ZNF816A基因插入编码变异与汉族人银屑病发病机制相关性研究

Psoriasis is a common chronic inflammatory skin disease which is difficult to effect a radical cure and affects physical and mental health of patients seriously. Our team analyzed the exome and targeted sequencing data of 10,727 Han psoriasis patients and 10,582 controls and found a new insertion variant c.552_553insAGT (p.Gly185Serfs*468) significantly associated with psoriasis(P=2.12×10-8). The results of immunohistochemistry suggests that the expression of the encoded protein is increased in psoriatic lesions. Zinc finger protein has important functions in gene regulatory and Zinc finger protein family has been reported to be associated with several types of cells. To confirm the association between the insertion variant of ZNF816A and psoriasis, we plan to validate the association between psoriasis and the variant using Sanger sequencing in 2,000 independent Han psoriasis patients and 2,000 matched controls, and plan to explore the functions of ZNF816A in the level of molecular levels using RT-PCR, Western blot, MTT, induced differentiation, apoptosis detection and siRNA interference to reveal the role of ZNF816A gene on the pathogenesis of psoriasis. It will lay a good foundation for predicting disease risk and provide basis for clinical diagnosis, therapy and new drug development of psoriasis based on ZNF816A.

银屑病是一种常见的慢性炎症性皮肤病,难以根治,严重危害患者身心健康。课题组前期在10727例汉族银屑病患者和10582例对照的大样本量外显子测和靶向测序数据分析发现ZNF816A基因新的插入变异c.552_553insAGT（p.Gly185Serfs*468）与汉族人群银屑病显著关联（P=2.12×10-8），免疫组化提示ZNF816A在银屑病皮损中的表达水平增高。锌指蛋白对基因调控起重要作用,目前锌指蛋白家族被发现与多种细胞的增殖、分化和凋亡相关。为明确该插入变异与银屑病发病的相关性，本课题拟选取独立的2000例汉族银屑病病例和2000例对照，并测序验证该插入变异，结合RT-PCR、Western blot、MTT、诱导分化、凋亡检测及RNA干扰等功能实验，在分子水平阐释ZNF816基因在银屑病发病机制中的作用，为基于ZNF816A的银屑病风险预测、临床诊疗和新药研发提供依据。

本项目基于团队前期外显子组测序和靶向测序数据，分析ZNF816A基因的插入变异c.552_553insAGT及ZNF816A基因与银屑病的关联。通过在中国人群5,422例病例和5,240例对照进行基因分型，联合前期高通量测序数据，结果发现该插入变异与银屑病关联达到全基因组显著水平，将该变异与ZNF816A基因已报道的变异位点进行条件分析，发现插入变异与银屑病的关联更显著。运用 SOAP-PopIndel 方法对32,043名中国汉族人的1,326个基因的外显子组序列数据进行深入筛选，在25个与银屑病相关的易感基因中鉴定出29个未报道的插入缺失变异。其中包括12个常见变异，9个低频变异和8个罕见变异，总计解释了约1.29% 的银屑病遗传度。 进一步分析证实LEKR1，AFF3，KIAA0319，，LMAN1，FLG，HERC5，RELN， AADACL2，CCDC66， ABCG2，ANXA7，SYTL2， ABO，GIPR，NCAPG，METTL1和FYCO1为未之前未报道的银屑病基因。 而其中部分插入缺失变异与已报道的基因 ERAP1，IFIH1， INPEP，UBLCP1，STAT3和 ERAP2有关，而位于GJB2基因和ZNF816A基因外显子上的插入缺失变异是既往未曾报道的独立关联，该研究进一步丰富了银屑病发生的遗传学机制，为银屑病的风险预测、临床诊疗和新药开发提供依据。同时证实了插入缺失变异在复杂疾病中的重要作用。