The Role of Interferon Regulatory Factor 5 in the Pathogenesis of SLE

摘要 系统性红斑狼疮(SLE)是一种自身免疫性疾病，约0.1%的患者 并导致相当大的发病率和寿命缩短。目前的治疗主要是基于非 特异性免疫抑制，通常只有部分有效，可能有严重的副作用。这个 迫切需要开发更具体、有效和更安全的治疗方法。基因的多态现象 转录因子干扰素调节因子5(IRF5)在人类遗传学研究中与 罹患系统性红斑狼疮和其他自身免疫性疾病的风险增加。IRF5在Toll-Like中的重要作用 受体信号。IRF5缺乏显著降低了许多小鼠模型的疾病严重程度 SLE。综上所述，抑制IRF5可能是治疗SLE的一种有效方法。这个 该项目的目标是获得有关IRF5功能和激活的详细信息，将导致更好的 了解系统性红斑狼疮发病的基本机制和潜在的新的治疗方法 抑制IRF5和寻找新的治疗靶点。这将通过以下方式完成：(1)确定IRF5- 特异性缺失红斑狼疮小鼠模型中介导疾病的表达细胞类型(S) 免疫细胞类型和评估缺失对疾病发展的影响；(2)确定 在疾病建立后删除IRF5可以逆转小鼠的疾病或阻止疾病的进展 (3)采用逆转录技术和CRISPR基因编辑互补的方法建立SLE模型 建立新的小鼠模型以确定狼疮所需的IRF5中的特定磷酸化位点(S) 体内致病机制和IRF5在体外原代免疫细胞中的功能和激活；(4)确定 与狼疮风险相关的IRF5基因多态性调节人类髓系细胞中IRF5的表达和功能 树突状细胞。这将通过从外周血中制造诱导多能干细胞(IPS)来实现 携带和不携带IRF5风险基因的健康志愿者单个核细胞分化 使用一种新的体外技术将iPS细胞转化为髓系树突状细胞。此外，为了最终确定 确定IRF5风险多态的影响，该多态将被引入到非 使用基因编辑和功能IRF5反应的多态iPS细胞将在同基因髓系中进行比较 除了感兴趣的风险多态外，在基因上相同的树突状细胞。

ABSTRACT Systemic lupus erythematosus (SLE) is an autoimmune disease that affects approximately 0.1% of the population and causes substantial morbidity and reduced lifespan. Current treatment is largely based on non- specific immunosuppression which is often only partially effective and may have serious side-effects. The development of more specific, effective and safer therapies is urgently needed. Polymorphisms in the transcription factor interferon regulatory factor 5 (IRF5) are strongly associated in human genetic studies with an increased risk of developing SLE and other autoimmune diseases. IRF5 plays an important role in Toll-like receptor signaling. Deficiency of IRF5 markedly reduces disease severity in a number of mouse models of SLE. Taken together, this suggests that IRF5 inhibition may be an effective therapeutic approach in SLE. The goal of this project is to obtain detailed information about IRF5 function and activation that will lead to a better understanding of the basic mechanisms underlying SLE pathogenesis and potentially to new approaches to inhibit IRF5 and the identification of novel therapeutic targets. This will be done by: (1) determining the IRF5- expressing cell type(s) responsible for mediating disease in a mouse model of SLE by deleting IRF5 in specific immune cell types and evaluating the effect of the deletion on disease development; (2) determining whether deletion of IRF5 after disease is established can reverse disease or prevent disease progression in a mouse model of SLE; (3) using the complementary approaches of retrogenic technology and CRISPR gene editing to create novel mouse models to determine the specific phosphorylation site(s) in IRF5 required for lupus pathogenesis in vivo and for IRF5 function and activation in primary immune cells ex vivo; (4) determining how the IRF5 polymorphisms associated with lupus risk modulate IRF5 expression and function in human myeloid dendritic cells. This will be done by making induced pluripotent stem (iPS) cells from peripheral blood mononuclear cells of healthy volunteers with and without the IRF5 risk polymorphisms and then differentiating the iPS cells into myeloid dendritic cells using a novel in vitro technique. In addition, in order to definitively determine the effect of the IRF5 risk polymorphisms, the polymorphisms will be introduced into non- polymorphic iPS cells using gene editing and functional IRF5 responses will be compared in isogenic myeloid dendritic cells that are genetically identical, except for the risk polymorphism of interest.