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

干扰素调节因子5在SLE发病机制中的作用

• 批准号: 9754572
• 负责人: IAN R RIFKIN
• 金额: $ 52万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-22 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9754572
• 关键词: Address; Affect; Alanine; Alleles; Autoimmune Diseases; B-Lymphocytes; Cell Line; Cells; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Dendritic Cells; Development; Dimerization; Disease; Disease Progression; Generations; Genes; Genetic Polymorphism; Genetic study; Goals; Hematopoietic stem cells; Human; Human Genetics; ITGAX gene; Immune; Immunosuppression; In Vitro; Individual; Interferon Activation; Interferons; Knowledge; Lead; Linkage Disequilibrium; Longevity; LoxP-flanked allele; Lupus; Mediating; Modeling; Morbidity - disease rate; Mus; Mutation; Myelogenous; Nuclear Translocation; Onset of illness; Pathogenesis; Peripheral Blood Mononuclear Cell; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; Population; Production; Receptor Signaling; Risk; Role; Series; Serine; Serine Phosphorylation Site; Severity of illness; Source; System; Systemic Lupus Erythematosus; T-Lymphocyte; TLR7 gene; Tamoxifen; Techniques; Technology; Therapeutic; Toll-like receptors; Treatment Efficacy; Variant; base; cell type; cellular transduction; cytokine; experimental study; healthy volunteer; human pluripotent stem cell; in vivo; in vivo Model; induced pluripotent stem cell; interest; lentiviral-mediated; loss of function; mouse model; new therapeutic target; novel; novel strategies; prevent; reconstitution; response; side effect; transcription factor

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.

抽象的 系统性红斑狼疮 (SLE) 是一种自身免疫性疾病，影响约 0.1% 的患者 人口并造成大量发病率和寿命缩短。目前的治疗主要基于非 特异性免疫抑制通常只能部分有效，并且可能有严重的副作用。这 迫切需要开发更特异、更有效和更安全的疗法。多态性在 转录因子干扰素调节因子 5 (IRF5) 在人类遗传学研究中与 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中发挥着重要作用 受体信号传导。 IRF5 缺陷可显着降低多种小鼠模型的疾病严重程度 系统性红斑狼疮。综上所述，这表明 IRF5 抑制可能是 SLE 的有效治疗方法。这 该项目的目标是获得有关 IRF5 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以及潜在的治疗新方法 抑制 IRF5 和新治疗靶点的鉴定。这将通过以下方式完成： (1) 确定 IRF5- 通过删除特定的 IRF5，在 SLE 小鼠模型中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) 判断是否 小鼠疾病发生后删除 IRF5 可逆转疾病或预防疾病进展 SLE模型； (3)利用逆转录技术和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 IRF5 中的特定磷酸化位点 体内发病机制以及离体初级免疫细胞中 IRF5 的功能和激活； (4) 确定如何 与狼疮风险相关的 IRF5 多态性调节人骨髓中 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（iPS）细胞来完成 具有和不具有 IRF5 风险多态性的健康志愿者的单核细胞，然后进行分化 使用一种新颖的体外技术将 iPS 细胞转化为骨髓树突状细胞。另外，为了最终 确定IRF5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 IRF5 反应的多态性 iPS 细胞将在同基因骨髓中进行比较 除了感兴趣的风险多态性之外，树突状细胞在遗传上是相同的。