Role of mitochondria in SLE and its cardiovascular complications

线粒体在 SLE 及其心血管并发症中的作用

• 批准号: 10274520
• 负责人: Jan Christian Lood
• 金额: $ 60.89万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-25 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10274520
• 关键词: Aerobic; Affinity; Anaerobic Bacteria; Antibodies; Anticardiolipin Antibodies; Antigens; Antiphospholipid Syndrome; Autoantibodies; Autoantigens; Autoimmunity; Biological Markers; Biology; Blood Platelets; Cardiolipins; Cardiovascular Diseases; Cardiovascular system; Cells; Cessation of life; Clinical; Coagulation Process; Complement; Complement 1q; Data; Development; Disease; Disease Management; Early treatment; Engineering; Excision; Exposure to; Flow Cytometry; Genome; Immune; Immune response; Impairment; In Vitro; Individual; Inflammation; Inflammatory; Life; Lupus; Mass Spectrum Analysis; Mediating; Methionine; Mitochondria; Mitochondrial Proteins; Modeling; Monitor; Mus; Organ; Outcome; Outcome Measure; Pathology; Pathway interactions; Patients; Phagocytosis; Phospholipids; Platelet Activation; Power Plants; Preventive measure; Preventive treatment; Process; Production; Prognostic Marker; Prokaryotic Cells; Property; Protein Biosynthesis; Reactive Oxygen Species; Research; Rheumatoid Arthritis; Risk; Role; Sampling; Signal Pathway; System; Systemic Lupus Erythematosus; Testing; Thrombosis; Thrombus; Venous Thrombosis; Work; base; cohort; complement system; cytokine; design; experimental study; extracellular; human disease; immunogenic; in vivo; in vivo Model; insight; mouse model; neutrophil; new therapeutic target; novel; novel marker; phosphoproteomics; prognostic; therapeutic target; venous thromboembolism

Summary: Upon cellular death, cells release damaged mitochondria, contributing to local inflammation. The overall aims of the present study are to investigate how mitochondria are cleared to avoid evoking an aberrant immune response, and to determine if autoantibodies targeting mitochondrial proteins have clinical utility in assessing development of thrombosis in patients with systemic lupus erythematosus (SLE). The premise of the study is that SLE patients have impaired clearance of mitochondria, promoting development of mitochondrial antibodies, inflammation and organ damage, including thrombosis. To investigate this we have three main aims. The first aim investigates how mitochondrial extrusion promotes autoimmunity towards mitochondrial protein antigens, attempting to define the exact target of novel main mitochondrial autoantibody, AMA-17, the processes involved in exposing the autoantigen in vitro, as well as potential therapeutic targets, e.g. mitochondrial ROS, involved in generating AMA-17 in vivo. The second aim investigates how anti-mitochondrial antibodies partake in thrombosis development. Using a large longitudinal SLE cohort (n=500), followed over 10 years, we will determine the capacity of AMA-17 to associate with and/or predict development of venous thrombosis. Affinity-purified AMA-17 antibodies will be tested for platelet activation and thrombus formation in vitro using flow cytometry, aggregometry and a cutting-edge model of engineered microvessels, as well as in vivo using a novel model of antibody-mediated venous thrombosis developed by Dr. Knight. Finally, in Aim 3, we will investigate underlying mechanisms involved in clearance of mitochondria, with an emphasis on the role of complement C1q and C3 to facilitate silent clearance. These studies will be done both in vitro using select isolated complement components, as well as in vivo using unique C1q and C3 deficient mice. Outcome measures include phagocytosis, cytokine production, and NET formation. Downstream signaling pathways involved in mitochondrial-mediated inflammation will be identified using mass spectrometry-based phosphoproteomics. In all, our study aims at identifying fundamental mechanisms regulating inflammation, thrombosis and autoimmunity in the context of human disease, with an emphasis on the role of the complement system in silent removal of mitochondria. We expect the proposed research to provide novel therapeutic targets disrupting the inflammatory and immunogenic properties of mitochondria, applicable for many diseases, including SLE and rheumatoid arthritis, as well as identify prognostic mitochondrial- derived biomarkers enabling early and preventive treatment of venous thrombosis.

摘要：细胞死亡后，细胞释放受损的线粒体，有助于局部 炎。本研究的总体目标是研究线粒体如何 清除以避免引起异常免疫反应，并确定自身抗体是否 靶向线粒体蛋白在评估血栓形成的发展方面具有临床实用性 系统性红斑狼疮 (SLE) 患者。该研究的前提是 SLE 患者 线粒体清除受损，促进线粒体抗体的形成， 炎症和器官损伤，包括血栓形成。为了调查这个问题，我们有三个主要的 目标。第一个目标是研究线粒体挤压如何促进自身免疫 线粒体蛋白抗原，试图定义新型主要线粒体的确切靶标 自身抗体，AMA-17，体外暴露自身抗原的过程，以及 潜在的治疗目标，例如线粒体 ROS，参与体内生成 AMA-17。 第二个目标是研究抗线粒体抗体如何参与血栓形成 发展。使用大型纵向 SLE 队列（n = 500）并跟踪 10 年以上，我们将 确定 AMA-17 关联和/或预测静脉发育的能力 血栓形成。将测试亲和纯化的 AMA-17 抗体的血小板活化和 使用流式细胞术、聚集测量法和尖端的血栓形成模型 工程微血管，以及体内使用抗体介导的静脉的新型模型 奈特博士开发的血栓形成。最后，在目标 3 中，我们将研究潜在的 涉及线粒体清除的机制，重点是补体的作用 C1q和C3便于无声通关。这些研究将在体外使用选择进行 分离补体成分，以及在体内使用独特的 C1q 和 C3 缺陷小鼠。 结果指标包括吞噬作用、细胞因子产生和 NET 形成。 参与线粒体介导的炎症的下游信号通路将是 使用基于质谱的磷酸蛋白质组学进行鉴定。总而言之，我们的研究目的是 确定调节炎症、血栓形成和自身免疫的基本机制 人类疾病的背景，重点是补体系统在沉默中的作用 去除线粒体。我们期望拟议的研究能够提供新的治疗靶点 破坏线粒体的炎症和免疫原性，适用于许多 疾病，包括系统性红斑狼疮和类风湿性关节炎，以及确定预后线粒体 衍生的生物标志物能够对静脉血栓形成进行早期和预防性治疗。