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

• 批准号: 9758932
• 负责人: Kathryn Alexandria Wierenga
• 金额: $ 3.58万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 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/9758932
• 关键词: Address; Agonist; Alveolar Macrophages; Anti-inflammatory; 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; 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; Address; Agonist; Alveolar Macrophages; Anti-inflammatory; 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; 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

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 该杂志在自身免疫性和炎症性疾病的发展中起着至关重要的作用。在这个亲 POSAL，我将解决饮食ω-3脂肪酸docosahexaenoic（DHA）在防御中的作用 由可呼吸的有毒物质二氧化硅（CSIO2）引起的炎症。以前，我们的实验室发现 补充DHA剂量依赖性地降低了CSIO2触发的自动IM的几种特征 狼疮型鼠标模型中的城市。该模型中系统注射开发的关键事件是 CSIO2诱导的肺泡巨噬细胞（AMPH）的毒性，涉及nlrp3 insperam- Masome和潜在IL-1细胞因子的释放。当前的文献和我的初步实验表明 DHA及其代谢产物（称为专业的预介介媒体（SPM））可能会减轻这种重新 赞助。 NLRP3炎性体的激活，在许多炎症和自身免疫中实施 条件，需要一个初始启动步骤，在此期间，NF-KB家族转录因子上调了inspram- 共体成分和Pro-IL-1细胞因子。抗炎G蛋白偶联受体（GPCR）具有 被鉴定出结合DHA或SPM并抑制NF-KB激活。但是，在体外阐明分子 AMPH中DHA保护的事件受单一小鼠可获得的细胞数量少（〜105）的限制。 为了解决这个问题，我使用了Max Planck Institute（MPI）细胞。 MPI细胞是一种自我更新的巨噬细胞系 由生活在GM-CSF补充培养基中的文化胎儿小鼠衍生而成，在表型上与 Amph。我的初步数据表明，IL-1细胞因子响应LPS酸化和CSIO2处理是 补充DHA。我建议补充DHA会增加细胞膜中的DHA 可以释放并代谢为SPM的MPI细胞。 Free DHA及其SPM可以激活抗激素 - 以自分泌或旁分泌方式的机械GPCR减弱NF-KB信号，我假设这是一种 玛丽的机制，它们可以防止CSIO2诱导的炎症。在AIM 1中，磷脂incorpo- 将测量DHA的评估，然后DHA对IL-1细胞因子释放和NF-KB激活的影响 评估。我还将用化学激动剂，拮抗剂和siRNA处理特定的GPCR，以验证它们的细胞 在DHA信号中的作用。在AIM 2中，将测量补充DHA的细胞的脂质代谢物谱。我 然后，将确定SPM抑制NF-KB激活和IL-1细胞因子释放的程度。最后，化学 ICAL拮抗剂和siRNA将用于研究拟议的SPM受体的参与。这些前 将在受支持的环境中进行必要资源以实现这些ob- 我的全面培训计划提供了个人和专业发展，这将有助于我 成为一名成功的独立研究员。