Ferroptosis as a Potential Mechanism of Blood-Induced Chondrocyte Cell Death

铁死亡作为血液诱导软骨细胞死亡的潜在机制

• 批准号: 10473766
• 负责人: Andy Jaehan Lee
• 金额: $ 4.66万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10473766
• 关键词: Address; Affect; Anatomy; Apoptosis; Autologous Transplantation; Biochemical; Biological Models; Biology; Biomedical Engineering; Biotechnology; Blood; Blood Cells; Blood Coagulation Factor; Blood Vessels; Blood coagulation; Blunt Trauma; Cartilage; Cell Death; Cells; Characteristics; Chondrocytes; Clinical; Coagulation Process; Coculture Techniques; Coupled; Degenerative polyarthritis; Development; Disease; Dose; Enzymes; Erythrocytes; Exposure to; Factor IX; Factor VIII; Fibroblasts; Fracture; Genetic Diseases; Glass; Hemarthrosis; Hemophilia A; Hemophilic Arthritis; Hemorrhage; Human; Hydroxyl Radical; Iatrogenesis; Immune system; Inflammatory; Infusion procedures; Intervention; Intracellular Accumulation of Lipids; Iron; Joints; Knee joint; Lead; Left; Link; Lipid Peroxides; Liquid substance; Mechanics; Mediating; Mesenchymal; Modernization; Molecular Biology Techniques; Monitor; Morbidity - disease rate; Necrosis; Operative Surgical Procedures; Outcome; Patients; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Physiological; Plasma; Play; Polyunsaturated Fatty Acids; Process; Property; Prophylactic treatment; Proteoglycan; Recombinants; Recurrence; Research Support; Role; Synovial Fluid; Synovial Membrane; Synovial joint; Synovitis; System; Therapeutic; Time; Tissues; Transplantation Surgery; Trauma; anterior cruciate ligament injury; articular cartilage; cartilage degradation; cytokine; effective therapy; experimental study; in vitro Model; inhibiting antibody; inhibitor; joint injury; joint loading; macrophage; metabolomics; novel; osteochondral tissue; prevent; response; tool

Project Summary Hemophilia is an X-linked genetic disorder that prevents blood from clotting normally due to a deficiency in either coagulation factor VIII or factor IX. Uncontrolled internal bleeding can occur during surgery and from blunt force trauma-induced breakage of blood vessels in the synovial membrane. Recurrent episodes of hemarthrosis lead to complete destruction of the articular cartilage, known as hemophilic arthropathy. While the exact mechanisms of blood-induced cartilage damage remain unclear, apoptosis is implicated as the primary form of regulated cell death (RCD) in chondrocytes due to elevated concentrations of pro-inflammatory cytokines. However, other forms of RCD may play roles in inducing damage to cartilage as well as the surrounding synovium. Ferroptosis is a recently discovered, iron-dependent, nonapoptotic form of RCD associated with excessive intracellular accumulation of lipid hydroperoxides formed from free hydroxyl radicals and polyunsaturated fatty acids. This proposal explores ferroptosis as a potential mechanism of joint tissue damage caused by excess intra-articular iron released from blood. In addition, ferroptosis inhibitors such as Ferrostatin-1, which prevent the formation of lipid hydroperoxides, are explored as potential therapeutics against blood-induced cell death. This new F31 proposal will fill a large gap in the current understanding of hemophilic arthropathy. Better understanding of the dose-response of blood exposure and duration may yield potential therapeutic windows of intervention that abrogate the sequela of joint bleeding. The role of physiologic joint loading and the synovium on blood-induced cartilage damage will also be addressed using a synovial joint model system coupled with modern bioengineering and molecular biology techniques (e.g., metabolomics). Isolating the effects of blood on cartilage, synovium, and their co-culture will inform new targets aimed at chondroprotection from joint bleeding. Hypothesis 1: Blood-induced cartilage damage is due in part to ferroptosis of articular chondrocytes. Specific Aim 1: A) Perform dose-response to blood on mechanical and biochemical properties of articular cartilage. Assess relative contribution of necrosis, apoptosis and ferroptosis in blood-induced cartilage damage. B) Study respective contribution of blood constituents to joint tissue damage and compare with ferroptosis inducers. C) Subject blood to fluid-induced shear and monitor erythrocyte viability and blood-cell related products in the synovial fluid. D) Assess ability of Ferrostatin-1 to mitigate blood-induced changes to cartilage. Hypothesis 2: Blood related cartilage damage is exacerbated by blood-induced changes to synovium. Specific Aim 2: Repeat Specific Aim 1 on synovium and cartilage-synovium co-culture. A) Perform reciprocating shear of synovium-on-cartilage and cartilage-on-cartilage. B) Perform conditioned media experiments that transfer media from reciprocal shear loading of cartilage-on-glass to synovium culture or synovium-on-glass to cartilage culture. Perform no-loading controls.

项目摘要 血友病是一种X连锁的遗传疾病，由于任何一种 凝结因子VIII或第IX因子。手术期间可能发生不受控制的内部出血和钝力 创伤引起的滑膜血管破裂。血肿铅的复发发作 为了彻底破坏关节软骨，称为血友病性关节病。而确切的机制 血液诱导的软骨损伤仍然不清楚，凋亡被认为是调节细胞的主要形式 由于促炎细胞因子浓度升高，软骨细胞的死亡（RCD）。但是，其他 RCD的形式可能在诱导软骨以及周围滑膜的损害方面发挥作用。铁凋亡 是一种最近发现的，铁依赖性的，非凋亡形式的RCD，与细胞内过多有关 由游离羟基自由基和多不饱和脂肪酸形成的脂质氢过氧化物的积累。这 提案探讨了铁铁作用，这是关节内造成的关节组织损伤的潜在机制 铁从血液中释放出来。此外，诸如Ferrostatin-1之类的铁凋亡抑制剂，以防止形成 脂质氢过氧化物被探讨为针对血液诱导的细胞死亡的潜在疗法。 这项新的F31提案将填补当前对血友人关节病的理解的巨大空白。更好的 了解血液暴露和持续时间的剂量反应可能会产生潜在的治疗窗口 消除关节出血的后遗症的干预措施。生理关节负荷和滑膜的作用 在血液诱导的软骨损伤上也将使用滑膜关节模型系统解决 现代生物工程和分子生物学技术（例如代谢组学）。隔离血液对 软骨，滑膜及其共培养物将为旨在通过关节出血的软骨保护的新目标提供信息。 假设1：血液诱导的软骨损伤部分归因于关节软骨细胞的铁tho虫。具体的 目标1：a）对关节软骨的机械和生化特性对血液进行剂量反应。 评估血液诱导的软骨损伤中坏死，细胞凋亡和铁凋亡的相对贡献。 b）研究 血液成分对关节组织损伤的各自贡献，并与铁凋亡诱导剂进行比较。 c） 血液受到流体诱导的剪切和监测红细胞的生存能力和与血细胞相关的产物的受试者。 滑液。 d）评估费罗斯替汀1减轻血液变化的能力。 假设2：血液诱导的滑膜变化加剧了与血液相关的软骨损伤。具体的 AIM 2：重复特定目标1对滑膜和软骨共培养。 a）执行往复剪切 仪式上的基团和软骨。 b）执行调节媒体的条件媒体实验 从软骨玻璃的相互剪切载荷到滑膜培养或玻璃滑膜到软骨培养物。 执行无加载控件。