调控狼疮病人I 型干扰素通路异常活化的关键分子的筛选鉴定及功能解析

Type I interferon (IFN) signaling has been documented as a central pathogenic pathway in systemic lupus erythematosus (SLE). Specific inhibition of this pathway has emerged as a core field of the development of new drugs for SLE. We previously identified “IFN signature”of lupus patients and took it as pharmacokinetic biomarker guiding the clinical drug trials for SLE therapy. In order to further elucidate the molecular mechanism of “IFN signature”expressed abnormly in SLE patients , we developed a high-throughput screening platform for functional genomics system to identify new type I IFN pathway regulators. Through screening and in vitro cell intervention experiment, we first found that downregulation of CDK1,which is a cell cycle regulatory molecule ,could significantly inhibit the activation of various levels of interferon pathway. Meanwhile,CDK1 was over-expressed in peripheral blood mononuclear cells (PBMCs) of SLE patients and was positively correlated with the type I IFN scores, suggesting that CDK1 may be responsible for abnormal activation of I IFN pathway in the SLE. On the basis of these study,by using various genomics and proteomics strategy, we will deeply and comprehensively elucidate the molecular mechanism of CDK1 regulation of IFN pathway.At the same time, by using peripheral blood cells of lupus patients and lupus mice model,we could observe the phenotype changing of SLE by intervention CDK1 expression in both vitro and in vivo expreriments. This study will help to reveal a specific molecular mechanism of the abnormal activation of the IFN pathway in SLE patients and and provide an important target for therapeutic intervention.

I型干扰素（IFN）通路异常活化在系统性红斑狼疮（SLE）发生发展中起关键作用，特异性阻断此通路是SLE新药研发的核心领域。我们前期鉴定了一组IFN下游基因可作为SLE病人分子分型和指导药物试验的基因组学标记物。为了进一步阐明SLE病人这组IFN下游基因活化的分子机理，我们建立了高通量功能基因组学的筛选平台系统鉴定新的IFN通路调控分子，首次发现了细胞周期调控分子CDK1可以在细胞水平影响IFN下游通路，又在狼疮病人外周血芯片中证实该基因差异表达且与干扰素积分正相关，提示CDK1在SLE病人IFN通路的异常活化中起重要作用。在此基础上本项目将利用多种功能基因组学及蛋白质组学策略探究CDK1调控IFN通路的分子机制，并利用SLE病人的外周血细胞和狼疮小鼠模型硏究体内外干预CDK1的表达对SLE表型的影响，从而揭示SLE病人IFN通路激活的分子机制并为特异性干预治疗提供靶点。

I型干扰素（IFN）通路异常活化在系统性红斑狼疮（SLE）发生发展中起关键作用,特异性阻断此通路是SLE新药研发的核心领域.我们建立了稳定高效的高通量筛选平台,筛选鉴定出一批可以调控一型干扰素通路的新的分子.结合大样本量的临床数据,我们挑选出在狼疮患者中差异表达的候选分子,检测候选分子与狼疮患者干扰素积分的相关性.我们发现CDK1在狼疮患者的外周血单个核细胞以及肾脏组织中相对于正常健康对照高表达,同时在狼疮患者的外周血以及常见的靶组织如皮肤组织和肾脏组织中,CDK1的表达与干扰素积分正相关.而在体外分子试验中,我们证明了CDK1是IFN信号通路的正向调控分子,CDK1可以影响IFN通路中关键的信号分子STAT1的磷酸化、影响干扰素诱导基因的顺式作用元件ISRE 的荧光报告基因活性,最终影响干扰素诱导基因的表达.为了探究CDK1在狼疮发病机制中的作用,我们使用CDK1的小分子化合物抑制剂,发现CDK1的小分子抑制剂可以抑制这些病理状态下的细胞中过度活化的I型干扰素通路。此外我们验证了多个参与狼疮I IFN通路异常活化的非编码RNA，发现mir130b、lncRNA RP11-2B6.2通过不同的表观遗传转录调控参与狼疮发病。本项目为I IFN在狼疮中的致病分子机制领域绘制了多元调控网络，提供了针对I IFN通路的多个可能新药靶点。