Roles of the Src-Family Kinases LynA and LynB in Macrophage Inflammatory Signaling

Src 家族激酶 LynA 和 LynB 在巨噬细胞炎症信号传导中的作用

• 批准号: 10308161
• 负责人: Tanya S. Freedman
• 金额: $ 4万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10308161
• 关键词: Adolescent; Affect; Arthritis; Autoimmune Diseases; Bacterial Infections; Bone Marrow; Cells; Childhood; Chronic; Chronic Childhood Arthritis; Clinic; Collaborations; Disease; Eye; Goals; Human; Hypersensitivity; Immune; Immune response; Immune system; Immunity; Immunology; Infection; Inflammation; Inflammatory; Inflammatory Arthritis; Innate Immune System; Joints; Laboratories; Liquid substance; Minnesota; Mus; Mycoses; Parents; Pathologic; Patients; Predisposition; Proteins; Regulatory Pathway; Research; Research Personnel; Rheumatology; Role; Sampling; Signal Pathway; Signal Transduction; Site; Stimulus; Symptoms; Synovial Fluid; Testing; United States; Universities; Up-Regulation; Wild Type Mouse; antimicrobial; bone; chronic pain; common treatment; knock-down; macrophage; mimetics; new therapeutic target; novel; novel therapeutic intervention; pathogen; response; side effect; src-Family Kinases; targeted treatment

ABSTRACT (from parent R01 AR073966) The autoimmune disease juvenile idiopathic arthritis (JIA), which causes joint, eye, and bone damage and chronic pain, is the most common chronic childhood disease in the United States. Joint fluid from JIA patients contains large numbers of macrophage immune cells, and macrophage secretion of inflammatory factors drives the disease. Current therapies for JIA block this inflammatory signaling but do not discriminate between disease- associated inflammatory signaling and the normal immune response to infection. Therefore, suppression of an- timicrobial immunity is a severe side effect of these existing therapies. We have discovered a signaling mecha- nism through the protein LynA that is required for hypersensitive inflammatory signaling by macrophages, but is not strictly required for pathogen recognition. We have now also generated the first LynA-/- mice and propose to test whether the LynA regulatory pathway has promise as a novel therapeutic target, to specifically suppress inflammatory macrophage signaling without suppressing the immune system generally. Our long-term goal is to develop new therapeutic approaches targeted specifically to pathological macrophages. To evaluate the trans- lational potential of our signaling discovery, we aim to test the role of LynA in isolated macrophages, in mouse inflammatory arthritis, in infection, and in samples from human JIA patients. More specifically, we propose to: 1) Examine the effect of LynA deletion in macrophages: We will use novel LynA-/- mice developed recently in our laboratory to test the independent contributions of LynA to signaling in bone-marrow-derived macrophages. We predict that LynA-/- macrophages will have blunted responses to small stimuli but normal responses to large, pathogen-mimetic stimuli. 2) Examine the role of LynA in mouse inflammatory arthritis and infection: We will assess the progression of inflammatory arthritis and fungal and bacterial infections in wild-type and LynA-/- mice. We will also analyze markers of macrophage function and test signaling responses in splenic and synovial mac- rophages to assess the role of LynA in inflammation. We predict that LynA-/- mice will develop less severe arthritis than wild-type mice but respond normally to infection. 3) Test inflammation-driven LynA upregulation and signal- ing in human macrophages: We hypothesize that, as in mice, inflammation in JIA-affected joints induces LynA upregulation in human macrophages. We will test whether synovial fluid from JIA patients induces LynA upreg- ulation and macrophage hypersensitivity and whether this pathological function is suppressed by knocking down LynA expression. We predict that JIA synovial fluid will induce upregulation of LynA in human macrophages, leading to hypersensitive signaling. This project will benefit from our expertise in macrophage signaling and close collaborations with the research-oriented Pediatric Rheumatology Clinic and other investigators in the Center for Immunology at the University of Minnesota. These studies will establish the feasibility of targeting LynA to de- velop safer, targeted treatment options for JIA and other macrophage-driven inflammatory diseases.

摘要(摘自家长R01 AR073966) 自身免疫性疾病幼年特发性关节炎(JIA)，导致关节、眼睛和骨骼损伤和 慢性疼痛，是美国最常见的儿童慢性疾病。JIA患者的关节液 含有大量巨噬细胞免疫细胞，并驱动巨噬细胞分泌炎症因子 这种疾病。目前对JIA的治疗方法阻断了这种炎症信号，但不区分疾病- 相关的炎症信号和对感染的正常免疫反应。因此，压制An- 抗微生物免疫是这些现有疗法的严重副作用。我们发现了一种信号机制-- NISM通过LYNA蛋白实现，该蛋白是巨噬细胞超敏炎症信号传递所必需的，但 病原体识别不是严格要求的。我们现在还培育了第一只Lyna-/-小鼠，并提议 测试Lyna调节通路是否有希望成为新的治疗靶点，以具体抑制 炎性巨噬细胞信号传导一般不会抑制免疫系统。我们的长期目标是 开发专门针对病理性巨噬细胞的新治疗方法。为了评估反式- 我们发现信号的潜在可能性，我们的目标是测试Lyna在分离的巨噬细胞中的作用，在小鼠 炎症性关节炎、感染和人类JIA患者的样本。更具体地说，我们建议：1) 检查Lyna缺失对巨噬细胞的影响：我们将使用我们最近开发的新的Lyna-/-小鼠 实验室测试Lyna对骨髓来源的巨噬细胞信号的独立贡献。我们 预测Lyna-/-巨噬细胞对小刺激的反应迟钝，但对大的刺激反应正常， 病原体模拟刺激。2)研究Lyna在小鼠炎症性关节炎和感染中的作用：我们将 评估炎性关节炎、真菌和细菌感染在野生型和Lyna-/-小鼠中的进展情况。 我们还将分析巨噬细胞功能的标志物，并测试脾和滑膜巨噬细胞的信号反应。 评估Lyna在炎症中的作用的噬菌体。我们预测Lyna-/-小鼠的关节炎不会那么严重。 比野生型小鼠更强，但对感染的反应正常。3)测试炎症驱动的Lyna上调和信号- 人巨噬细胞中的ING：我们假设，就像在小鼠中一样，JIA感染的关节中的炎症导致Lyna 人巨噬细胞表达上调。我们将测试JIA患者的滑液是否诱导Lyna Upreg- 细胞生成和巨噬细胞超敏反应以及击倒是否抑制了这一病理功能 Lyna表达式。我们预测JIA滑液将诱导人巨噬细胞Lyna表达上调， 导致过度敏感的信号。这个项目将受益于我们在巨噬细胞信号传递和关闭方面的专业知识 与以研究为导向的儿科风湿病诊所和中心的其他研究人员合作 明尼苏达大学的免疫学博士。这些研究将确定将Lyna作为目标的可行性，以消除 为JIA和其他巨噬细胞驱动的炎症性疾病提供更安全、有针对性的治疗选择。