Omega-3 Fatty Acid Suppression of Silica-induced Inflammasome Activation in a Novel Alveolar Macrophage Model

Omega-3 脂肪酸在新型肺泡巨噬细胞模型中抑制二氧化硅诱导的炎症小体激活

基本信息

批准号：
9926082
负责人：
Kathryn Alexandria Wierenga
金额：
$ 3.66万
依托单位：
MICHIGAN STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-16 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9926082
关键词：
Address Agonist Alveolar Macrophages Anti-Inflammatory Agents Attenuated Autoimmune Diseases Autoimmune Process Autoimmunity Binding Cell Death Cell Line Cell Nucleus Cell membrane Cells Characteristics Chemicals Chronic Complex Data Development Diet Dietary Fats Dietary Supplementation Disease Docosahexaenoic Acids Docosahexaenoic acid supplementation Dose Dust Environment Enzymes Etiology Event Exposure to Family Fatty Acids Fetal Liver Fish Oils G-Protein-Coupled Receptors Genes Goals Granulocyte-Macrophage Colony-Stimulating Factor Human Immune In Vitro Inflammasome Inflammation Inflammation Mediators Inflammatory Inhalation Innate Immune Response Innate Immune System Institutes Interleukin-1 Interleukin-1 beta Intervention Knowledge Laboratories Lead Learning Link Lipids Lipoxygenase Literature Liver Lung Lung diseases Lupus Measures Mediating Mediator of activation protein Mission Modeling Molecular Mus NF-kappa B National Institute of Environmental Health Sciences Nuclear Translocation Omega-3 Fatty Acids Patients Phenotype Phospholipids Planet Earth Play Pre-Clinical Model Quartz Research Personnel Resources Role Signal Pathway Signal Transduction Silicon Dioxide Small Interfering RNA Supplementation System Systemic Lupus Erythematosus Testing Toxic Environmental Substances Toxic effect Toxicant exposure Training Unsaturated Fatty Acids Up-Regulation airway inflammation attenuation autocrine crystallinity cytokine disability experimental study fetal grasp human disease improved inhibitor/antagonist knock-down lipid mediator lung injury lupus prone mice macrophage man mouse model novel paracrine prevent receptor response self-renewal supportive environment tool toxicant transcription factor ward

项目摘要

ABSTRACT The exposome plays a critical role in the development of autoimmune and inflammatory diseases. In this pro- posal, I will address the role of the dietary ω-3 fatty acid docosahexaenoic acid (DHA) in protecting against inflammation induced by the respirable toxicant crystalline silica (cSiO2). Previously, our laboratory found that supplementation with DHA dose-dependently decreased levels of several features of cSiO2-triggered autoim- munity in a lupus-prone mouse model. A key event in the development of systemic inflammation in this model is cSiO2-induced toxicity of the alveolar macrophage (AMph), which involves activation of the NLRP3 inflam- masome and release of potent IL-1 cytokines. The current literature and my preliminary experiments suggest that DHA and its metabolites, known as specialized proresolving mediators (SPMs), may attenuate this re- sponse. Activation of the NLRP3 inflammasome, which is implicated in many inflammatory and autoimmune conditions, requires an initial priming step, during which NF-kB family transcription factors upregulate inflam- masome components and pro-IL-1 cytokines. Anti-inflammatory G-protein coupled receptors (GPCRs) have been identified that bind DHA or SPMs and inhibit NF-kB activation. However, in vitro elucidation of the molecular events of DHA protection in AMph is limited by the low number of cells attainable from a single mouse (~105). To address this, I used Max Planck Institute (MPI) cells. MPI cells are a self-renewing macrophage cell line derived by culturing fetal mouse livers in GM-CSF-supplemented medium and are phenotypically similar to AMph. My preliminary data show that IL-1 cytokine release in response to LPS-priming and cSiO2 treatment is attenuated by DHA supplementation. I propose that DHA supplementation increases DHA in the cell membrane of MPI cells, which can be released and metabolized to SPMs. Free DHA and its SPMs can activate anti-inflam- matory GPCRs in an autocrine or paracrine manner to attenuate NF-kB signaling, which I hypothesize is a pri- mary mechanism by which they protect against cSiO2-induced inflammation. In Aim 1, the phospholipid incorpo- ration of DHA will be measured, and then effects of DHA on IL-1 cytokine release and NF-kB activation will be assessed. I will also treat cells with chemical agonists, antagonists, and siRNA for specific GPCRs to verify their role in DHA signaling. In Aim 2, the lipid metabolite profile of cells supplemented with DHA will be measured. I will then determine the extent to which SPMs suppress NF-kB activation and IL-1 cytokine release. Lastly, chem- ical antagonists and siRNA will be used to investigate the involvement of proposed SPM receptors. These ex- periments will be performed in a supportive environment with the necessary resources to accomplish these ob- jectives. My comprehensive training plan provides personal and professional development, which will assist me in becoming a successful independent researcher.

抽象的暴露组在自身免疫性疾病和炎症性疾病的发展中起着至关重要的作用。接下来，我将讨论饮食中的 ω-3 脂肪酸二十二碳六烯酸 (DHA) 在预防此前，我们的实验室发现，由可吸入有毒物质结晶二氧化硅（cSiO2）引起的炎症。补充 DHA 可以剂量依赖性地降低 cSiO2 引发的自身免疫性多种特征的水平在易患狼疮的小鼠模型中，该模型中全身炎症发展的一个关键事件是。 cSiO2 诱导的肺泡巨噬细胞 (AMph) 毒性，涉及 NLRP3 炎症的激活目前的文献和我的初步实验表明。 DHA 及其代谢物（称为专门的促消退介质 (SPM)）可能会减弱这种再分解作用。 NLRP3 炎症小体的激活，与许多炎症和自身免疫性疾病有关。条件下，需要一个初始启动步骤，在此期间 NF-kB 家族转录因子上调炎症质体成分和前 IL-1 细胞因子具有抗炎 G 蛋白偶联受体 (GPCR)。已被鉴定结合 DHA 或 SPM 并抑制 NF-kB 激活，然而，该分子的体外阐明。 AMph 中 DHA 保护的事件受到单只小鼠可获得的细胞数量较少 (~105) 的限制。为了解决这个问题，我使用了马克斯普朗克研究所 (MPI) 细胞，MPI 细胞是一种自我更新的巨噬细胞系。通过在添加 GM-CSF 的培养基中培养胎鼠肝脏而获得，并且表型与 AMph。我的初步数据表明，LPS 引发和 cSiO2 处理后 IL-1 细胞因子的释放是我建议补充 DHA 可以增加细胞膜中的 DHA。 MPI 细胞可以释放并代谢为游离 DHA，其 SPM 可以激活抗炎作用。以自分泌或旁分泌的方式激活 GPCR 来减弱 NF-kB 信号传导，我认为这是一个优先因素。在目标 1 中，磷脂结合物可以防止 cSiO2 引起的炎症。测量 DHA 的比例，然后计算 DHA 对 IL-1 细胞因子释放和 NF-kB 激活的影响我还将用化学激动剂、拮抗剂和 siRNA 处理细胞以获得特定的 GPCR，以验证其效果。在目标 2 中，将测量补充 DHA 的细胞的脂质代谢谱。然后将确定 SPM 抑制 NF-kB 激活和 IL-1 细胞因子释放的程度。拮抗剂和 siRNA 将用于研究所提出的 SPM 受体的参与情况。实验将在具有必要资源的支持性环境中进行，以实现这些目标我的综合培训计划提供个人和专业发展，这将帮助我成为一名成功的独立研究员。