Autophagic regulation of inflammasome-mediated hyperactive state in living macrophages

活体巨噬细胞中炎症小体介导的过度活跃状态的自噬调节

• 批准号: 10449254
• 负责人: Venkat Giri Magupalli
• 金额: $ 19.28万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-12 至 2022-11-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10449254
• 关键词: Acute; Address; Antibodies; Autophagocytosis; Autophagosome; Binding; Biological; Biological Assay; CASP1 gene; COVID-19; Caspase; Cell Death; Cell membrane; Cells; Chemosensitization; Chronic Disease; Clinical; Colitis; Complex; Coupled; Cystic Fibrosis Transmembrane Conductance Regulator; Dynein ATPase; Family Dasypodidae; Genes; HDAC6 gene; Hematopoietic; Host Defense; Hyperactivity; Immune response; Infection; Inflammasome; Inflammation; Inflammatory; Injections; Interleukin-1; Interleukin-1 beta; Interleukin-18; Interleukins; Lead; Leucine-Rich Repeat; Ligands; Light; Link; Lipids; Lipopolysaccharides; Lysosomes; Lytic; Macrophage activation syndrome; Mediating; Membrane; Membrane Lipids; Microtubule-Organizing Center; Microtubules; Mitochondria; Mus; Nigericin; Nucleotides; Outcome; Pathway interactions; Peptide Hydrolases; Pericentriolar Regions; Phenotype; Phosphorylcholine; Process; Proteins; Publishing; Regulation; Reporting; Role; Science; Signal Transduction; Sodium Dextran Sulfate; Sterility; Stimulus; System; Time; Toll-like receptors; Yang; Yin; cellular imaging; cytokine; cytokine release syndrome; human disease; live cell imaging; macrophage; misfolded protein; oxidation; pathogen; presenilin-1; protein aggregation; recruit; repaired; response; retrograde transport; tissue injury

Abstract Nucleotide-binding domain (NBD) and leucine-rich repeat (LRR)-containing proteins (NLRs) assemble into functional supramolecular organizing centers called the inflammasomes, which serve as an interface of host defense against pathogens or other types of danger. Canonical inflammasomes activate caspase-1, an inflammatory protease that upon activation, triggers two important biological responses: 1) processing of cytokine interleukin-1b (IL-1b), and 2) formation of plasma membrane gasdermin D pores to potentiate inflammatory cell death (pyroptosis). Cytokine secretion and pyroptosis are coupled under a steady-state condition, as the GSDMD pores on the plasma membrane, which permeates IL-1b, can also compromise the cell membrane integrity leading to the lytic cell death. Interestingly, in a hyperactive state following inflammasome activation, cells show sustained cytokine secretion but retain viability, even though this process is dependent on GSDMD pores formed on the plasma membrane. The underlying mechanisms which recalibrate GSDMD to only sustain the release of the interleukin, but not pyroptosis, are poorly understood. Autophagy is regarded as an important regulator of inflammation. In our recent studies, we found that inflammasomes are regulated by an aggresome-like mechanism that promotes inflammasome assembly (Yang) but also induces autophagy to dampen caspase-1 processing and IL-b secretion (Yin) (Magupalli et al., Science, 2020). Our studies are consistent with previous reports showing that mice lacking the key autophagic gene, Atg16L1, in hematopoietic cells, were highly susceptible to dextran sulphate sodium-induced acute colitis, with the secretion of a high amount of IL-1b and IL-18 cytokines. In addition, these inflammatory phenotypes were alleviated by injection of anti-IL-1b and IL-18 antibodies in mice. Collectively, our and other published results demonstrate that autophagy directly regulates the extent to which inflammasomes activate downstream signals. However, it is unclear if autophagy can regulate the switch between a hyperactive state and a pyroptotic state of macrophages. In this application, we will investigate the potential link between autophagy and macrophage hyperactivation using inflammasome assays and cellular imaging.

摘要 核苷酸结合域(NBD)和富含亮氨酸重复序列(LRR)的蛋白质(NLR)组装成 功能性超分子组织中心称为炎性小体，它充当宿主的接口 对病原体或其他类型的危险的防御。典型炎症体激活caspase-1和 一种炎性蛋白水解酶，一旦激活，会触发两种重要的生物反应：1)细胞因子的处理 白介素1b(IL-1b)，以及2)形成质膜Gasdermin D孔，以增强炎症细胞 死亡(上睑下垂)。细胞因子的分泌和下垂是在稳态条件下耦合的，如GSDMD 渗透IL-1b的质膜上的小孔也会损害细胞膜的完整性。 导致溶细胞死亡。有趣的是，在炎症体激活后的过度活跃状态下，细胞显示 持续的细胞因子分泌，但保持活力，即使这一过程依赖于GSDMD毛孔的形成 在质膜上。重新校准GSDMD以仅维持GSDMD释放的潜在机制 白介素症，但不是上睑下垂，人们知之甚少。 自噬被认为是炎症的重要调节因子。在我们最近的研究中，我们发现 炎症体由一种类似侵袭体的机制调节，促进炎症体组装(杨) 而且还诱导自噬以抑制caspase-1的处理和IL-b的分泌(阴)(Magupalli等人，科学， 2020)。我们的研究与之前的报告一致，这些报告表明，缺乏关键自噬基因的小鼠， 在造血细胞中，Atg16L1对葡聚糖硫酸钠诱导的急性结肠炎高度敏感， 分泌大量IL-1b 和IL-18细胞因子。此外，这些 炎症表型为 小鼠注射抗IL-1b和IL-18抗体缓解。总体而言，我们和其他已公布的结果 证明自噬直接调节炎症小体激活下游信号的程度。 然而，目前尚不清楚自噬是否可以调节在过度活跃状态和嗜热性状态之间的切换。 巨噬细胞。 在这个应用中，我们将研究自噬和巨噬细胞之间的潜在联系。 使用炎症体分析和细胞成像的过度激活。