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，影响约0.1%的 这一现象严重影响了人口，并导致发病率高和寿命缩短。目前的治疗主要是基于非- 特异性免疫抑制，其通常仅部分有效且可能具有严重的副作用。的 迫切需要开发更特异、有效和更安全的疗法。多态性 转录因子干扰素调节因子5（IRF 5）在人类遗传学研究中与以下密切相关： SLE和其他自身免疫性疾病的风险增加。IRF 5在类Toll中起着重要作用 受体信号IRF 5的缺乏显著降低了许多小鼠模型的疾病严重程度， SLE。总之，这表明IRF 5抑制可能是SLE的有效治疗方法。的 本项目的目标是获得有关IRF 5功能和激活的详细信息，这将导致更好的 了解SLE发病机制的基本机制，并有可能找到新的方法， 抑制IRF 5和鉴定新的治疗靶点。这将通过以下方式实现：（1）确定IRF 5- 在SLE小鼠模型中通过特异性缺失IRF 5来表达负责介导疾病的细胞类型， 免疫细胞类型并评估缺失对疾病发展的影响;（2）确定是否 在疾病建立后缺失IRF 5可以逆转疾病或预防小鼠的疾病进展 SLE模型;（3）使用逆转录技术和CRISPR基因编辑的互补方法， 创建新的小鼠模型，以确定狼疮所需的IRF 5中的特异性磷酸化位点 （4）确定IRF 5在体内的发病机制以及在离体的原代免疫细胞中的功能和活化; 与狼疮风险相关的IRF 5多态性调节人骨髓中IRF 5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 具有和不具有IRF 5风险多态性的健康志愿者的单核细胞，然后区分 使用一种新的体外技术将iPS细胞转化为骨髓树突状细胞。此外，为了明确 确定IRF 5风险多态性的影响，将多态性引入非 使用基因编辑和功能性IRF 5应答的多态性iPS细胞将在同基因骨髓中进行比较。 树突状细胞是遗传上相同的，除了感兴趣的风险多态性。

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.