Investigating monocyte activation pathways in lupus

研究狼疮中的单核细胞激活途径

• 批准号: 10731104
• 负责人: Insoo Kang
• 金额: $ 36.85万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-19 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10731104
• 关键词: Address; Affect; Antibodies; Antigen-Antibody Complex; Autoantibodies; Autoantigens; Biological Assay; CASP1 gene; Caspase; Cell Death; Cell Death Induction; Cells; Characteristics; Clinical; Data; Dendritic Cells; Disease; Exposure to; Frequencies; Goals; Human; IL18 gene; Immune response; Inflammasome; Inflammation; Inflammatory; Interferon alpha; Interleukin-1 beta; Ku70 protein; Link; Lupus; Mediating; Membrane; Migration Inhibitory Factor; Molecular; Mus; Natural Immunity; Nuclear; Organ; Pathogenesis; Pathogenicity; Pathologic; Pathway interactions; Patients; Pattern; Phosphotransferases; Production; Protein Kinase Interaction; Publishing; RIPK1 gene; RIPK3 gene; Reporting; Role; Serology; Small Nuclear Ribonucleoproteins; Systemic Lupus Erythematosus; TNF gene; Testing; Text; Treatment Efficacy; U1 Small Nuclear Ribonucleoprotein; adaptive immunity; anti-dsDNA antibodies; chemokine; cytokine; ds-DNA; high dimensionality; improved; inhibitor; lupus prone mice; mixed lineage kinase 3; monocyte; novel strategies; response; single cell analysis; single-cell RNA sequencing; synergism; tissue injury

Project Summary/Abstract: The pathologic hallmarks of systemic lupus erythematosus (SLE or lupus) are altered immune responses to nuclear autoantigens with autoantibody production and subsequent tissue injury. Studies have suggested a pathogenic role for innate immunity in lupus. Plasmacytoid dendritic cells and monocytes (MO) of innate immunity recognize ssRNA of the self antigen U1-small nuclear ribonucleoprotein (U1-snRNP) and dsDNA via TLRs 7/8 and 9, respectively, leading to the production of IFN-α, IL-1β and IL-18 which are linked to lupus pathogenesis and clinical manifestations. Regulated cell death can occur in distinct forms including pyroptosis and necroptosis. Pyroptosis and necroptosis cause inflammation by releasing inflammatory molecules such as cytokines, chemokines, and damage-associated molecular patterns (DAMPs). Pyroptosis is induced by the activation of the caspase-1-containing inflammasomes that cleave pro-IL-1β and - IL18 into IL-1β and IL-18, as well as gasdermin D (GASDMD) into the active form gasdermin D amino-terminal fragment (GASDMDN), the essential molecule for membrane pore formation in pyroptosis. Necroptosis that occurs independently of caspases is regulated by TNF-α, receptor-interacting protein kinases 1 and 3 (RIPK1 and RIPK3), and downstream substrate pseudokinase mixed-lineage kinase domain-like (MLKL), the critical molecule for membrane disruption with the release of inflammatory molecules in necroptosis. RIPK3 and MLKL can also activate the NLRP3 inflammasome with IL-1β release, raising the possible interface of pyroptosis and necroptosis. Studies, including our own, support the implication of the inflammasome activation, pyroptosis, and necroptosis in lupus. We showed IL-1β and IL-18 production from human MO exposed to lupus U1-snRNP/anti- U1-snRNP antibody (Ab) or dsDNA/anti-dsDNA Ab immune complex (snRNP IC or dsDNA IC, also lupus IC refers to both) through the NLRP3 inflammasome activation which is enhanced by the cytokine macrophage migration inhibitory factor (MIF). However, there is a substantial scientific gap in our understanding of how pyroptic and necroptic pathways become activated, interface, and synergize to promote inflammation in lupus at molecular levels with contributing to unique clinical manifestations. Thus, we will test the hypotheses that lupus IC activate pyroptic and necroptic pathways in MO with synergistic interface in promoting inflammation and tissue injury and that the activation of such pathways alters in lupus contributing to clinicopathologic manifestations. The goal of the proposal is to address this hypothesis with: 1) Aim 1. Elucidate the mechanism and significance of the activation and interface of pyroptic and necroptic molecules in lupus; 2) Aim 2. Elucidate the “functional” alteration in monocytes that enhances pyroptosis- and necroptosis-mediated inflammation in lupus patients; and 3) Aim 3. Elucidate the role of macrophage migration inhibitory factor in promoting inflammation in lupus via regulating pyroptic and necroptic molecules. Our study would be highly informative in understanding lupus pathogenesis as well as in developing new approaches in evaluating and treating lupus patients.

项目摘要/摘要：系统性红斑狼疮（SLE 或狼疮）的病理特征是 通过自身抗体的产生和随后的组织损伤改变了对核自身抗原的免疫反应。 研究表明先天免疫在狼疮中具有致病作用。浆细胞样树突状细胞和 先天免疫的单核细胞（MO）识别自身抗原U1的ssRNA-小核核糖核蛋白 (U1-snRNP) 和 dsDNA 分别通过 TLR 7/8 和 9 产生 IFN-α、IL-1β 和 IL-18 与狼疮的发病机制和临床表现有关。受调控的细胞死亡可以发生在不同的 形式包括焦亡和坏死性凋亡。焦亡和坏死性凋亡通过释放引起炎症 炎症分子，例如细胞因子、趋化因子和损伤相关分子模式 (DAMP)。 细胞焦亡是由含有 caspase-1 的炎症小体的激活引起的，该炎症小体会裂解 pro-IL-1β 并 - IL18 转化为 IL-1β 和 IL-18，以及gasdermin D (GASDMD) 转化为活性形式gasdermin D 氨基末端 片段（GASDMDN），焦亡中膜孔形成的必需分子。坏死性凋亡 独立于 caspase 发生，受 TNF-α、受体相互作用蛋白激酶 1 和 3 (RIPK1 和 RIPK3），以及下游底物假激酶混合谱系激酶结构域（MLKL），关键 坏死性凋亡中膜破坏并释放炎症分子的分子。 RIPK3 和 MLKL 还可以激活 NLRP3 炎症小体并释放 IL-1β，从而提高焦亡和细胞焦亡的可能界面 坏死性凋亡。包括我们自己在内的研究支持炎症小体激活、细胞焦亡和 狼疮坏死性凋亡。我们展示了暴露于狼疮 U1-snRNP/抗-的人类 MO 产生的 IL-1β 和 IL-18 U1-snRNP 抗体 (Ab) 或 dsDNA/抗 dsDNA Ab 免疫复合物（snRNP IC 或 dsDNA IC，还有狼疮 IC 指两者）通过细胞因子巨噬细胞增强的 NLRP3 炎性体激活 迁移抑制因子（MIF）。然而，我们对于如何实现这一点的理解存在着巨大的科学差距。 热视通路和坏死视通路被激活、相互作用并协同作用，以促进狼疮的炎症 分子水平有助于独特的临床表现。因此，我们将检验以下假设：狼疮 IC 通过协同界面激活 MO 中的焦视和坏死视通路，促进炎症和组织 狼疮中这些途径的激活发生变化，导致临床病理表现。 该提案的目标是解决这一假设： 1) 目的 1. 阐明机制和意义 狼疮中焦视分子和坏死分子的激活和界面； 2) 目标 2. 阐明“功能性” 单核细胞的改变增强了狼疮患者焦亡和坏死性凋亡介导的炎症；和 3) 目标 3. 阐明巨噬细胞迁移抑制因子在促进狼疮炎症中的作用 调节热光和坏死分子。我们的研究对于了解狼疮将提供非常丰富的信息 发病机制以及开发评估和治疗狼疮患者的新方法。