线性泛素化缺陷和K63泛素化缺陷导致自身炎症疾病的分子机制

OTULIN和A20是重要的去线性和去K63泛素化酶，在炎症和免疫中有重要调控作用。申请人首次发现了OTULIN突变和A20突变导致的单基因遗传病，开启了对OTULIN和A20在人体中作用的研究。申请人之前研究发现了OTULIN缺失和A20缺失对NF-kB通路的影响。然而，OTULIN缺陷（Otulipenia）和A20缺陷（HA20）病人的表型截然不同，暗示还有更多的信号通路被调控。申请人初步的结果提示IFN信号通路在Otulipenia和HA20的致病机制中也起到关键的作用。本课题将通过IFN报告基因检测，Nanostring，RNA测序，RealtimePCR，免疫共沉淀，western blot等实验研究HA20和Otulipenia的IFN的信号通路的变化。发现OTULIN和A20在IFN通路上新的底物，研究OTULIN和A20突变对IFN信号通路的调控差异，揭示病人的致病原理。

OTULIN and A20 are the most important linear deubiqutinase and K63 deubiquitinase, both of which play critical roles in the inflammation and immunity. The applicant firstly identified mutant OTULIN and A20 causing monogenic inherited disorders，which opened a new area of investigating the regulation mechanisms of OTULIN and A20 in human. The applicant demonstrated that OTULIN defect and A20 defect resulted in the upregulation of NF-kB. However, OTULIN defect (Otulipenia) and A20 defect (HA20) patients have totally different clinical manifestations, indicating additional signaling pathways are involved. The primary data showed that IFN signaling may also play important roles in the pathophysiology. The purposed project plan to use IFN luciferase reporter assay, Nanostring, RNA seq, Realtime PCR, immunoprecipitation, western blot methods to study the regulation of IFN signaling in HA20 and Otulipenia, identify the new substrates of OTULIN and A20 in the IFN signaling pathway, compare the differences of OTULIN defect and A20 defect in affecting the IFN signaling pathway, and better understand the pathogenesis of Otulipenia and HA20.