Soluble adenylyl cyclases in lung endothelial tauopathy

肺内皮tau蛋白病中的可溶性腺苷酸环化酶

• 批准号: 10636060
• 负责人: Troy Stevens
• 金额: $ 49.68万
• 依托单位: UNIVERSITY OF SOUTH ALABAMA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-20 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10636060
• 关键词: Acute Lung Injury; Address; Adenylate Cyclase; Alveolar; Bacteria; Bacterial Infections; Bacterial Pneumonia; Binding; Bioenergetics; Blood; Blood Vessels; Blood capillaries; Bronchoalveolar Lavage Fluid; Capillary Permeability; Cells; Cerebrospinal Fluid; Clinical; Communication; Consumption; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cyclic GMP; Cyclic Nucleotides; Data; Dedications; Dissociation; Edema; Endothelial Cells; Endothelium; Enzymes; Functional disorder; Gases; Generations; Homologous Gene; Hydrolase; Impairment; Infection; Interleukin-1 Receptors; Knowledge; Lung; Mammalian Cell; Membrane; Microtubules; Morbidity - disease rate; Niacinamide; Nicotinamide adenine dinucleotide; Organ; Patients; Peripheral; Permeability; Phosphorylation; Pneumonia; Process; Production; Protein Kinase; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Recovery; Role; Signal Transduction; Site; Sterility; Stimulus; Structure; Tauopathies; Tertiary Protein Structure; Testing; Traction; Type III Secretion System Pathway; Variant; Work; brain parenchyma; cytotoxic; endothelial repair; exoenzyme; innate immune mechanisms; lung injury; lung repair; metabolomics; microorganism; mortality; mutant; novel; pathogen; prevent; repaired; response; tau Proteins; tau-1

PROJECT SUMMARY/ABSTRACT The alveolar-capillary membrane facilitates efficient gas exchange while maintaining a restrictive permeability barrier. Pseudomonas aeruginosa infection disrupts the alveolar-capillary barrier leading to exudative edema and impaired oxygenation. P. aeruginosa utilizes a type III secretion system and its effectors to disrupt barrier integrity. In particular, the exoenzyme Y is introduced into lung endothelium, where it acquires nucleotidylyl cyclase activity and produces cGMP, cAMP, and cUMP. These cyclic nucleotide monophosphates activate protein kinase A resulting in endothelial tau phosphorylation, tau dissociation from microtubules, and microtubule breakdown, which collectively hinders repair following infection. Phosphorylated tau is released from endothelium as cytotoxic variants that contribute to lung injury. The signaling mechanisms used by exoenzyme Y to produce cytotoxic tau is incompletely understood, yet cUMP is produced at especially high concentrations and the cUMP signal parallels the generation of cytotoxic tau. Elevations in cUMP are sufficient to promote the production of cytotoxic tau variants. Our preliminary data demonstrate that the exoenzyme Y-induced cUMP signal also decreases endothelial nicotinamide adenine dinucleotide (NAD+) and increases nicotinamide, the product of NAD+ hydrolase activity, which may impair recovery following infection. Lung endothelium expresses sterile alpha and TIR motif containing 1 (SARM1), the only TIR (Toll/Interleukin-1 Receptor) domain protein in mammalian cells that possesses NAD+ hydrolase activity. Recent studies revealed a SARM1 bacterial homologue is directly activated by cUMP as an essential innate immune mechanism. While our studies illustrate an important role for cUMP in the endothelial cell response to infection, how exoenzyme Y generates the cUMP that leads to tau phosphorylation and production of cytotoxic tau variants, and how cUMP lowers NAD+ while hindering endothelial cell repair remains poorly understood. To address this knowledge gap in a rigorous way, this project tests the hypothesis that the P. aeruginosa exoenzyme Y generates cUMP, which contributes to the tau phosphorylation, microtubule breakdown, and SARM1-dependent NAD+ hydrolase activity that causes lung injury and hinders repair.

项目摘要/摘要 肺泡-毛细血管膜在保持限制性通透性的同时促进有效的气体交换。 障碍。铜绿假单胞菌感染破坏肺泡-毛细血管屏障导致渗出性水肿 以及氧合功能受损。铜绿假单胞菌利用III型分泌系统及其效应器来破坏屏障。 正直。特别是，外切酶Y被引入肺内皮细胞，在那里它获得核苷酰 环化酶活性，并产生cGMP、cAMP和CUMP。这些环核苷酸单磷酸盐激活 蛋白激酶A导致内皮细胞tau磷酸化，tau从微管和微管解离 崩溃，这共同阻碍了感染后的修复。磷酸化的tau从 内皮细胞是导致肺损伤的细胞毒性变种。胞外酶使用的信号机制 产生细胞毒性tau的机制尚不完全清楚，但ccMP在特别高的浓度下产生。 而CUMP信号与细胞毒性tau的产生是平行的。CUMP中的高程足以促进 生产细胞毒性tau变异体。我们的初步数据表明，胞外酶Y诱导的CUMP Signal还会减少内皮细胞烟酰胺腺嘌呤二核苷酸(NAD+)，增加烟酰胺， NAD+水解酶活性的产物，这可能会损害感染后的恢复。肺内皮细胞表达 不育的α和TIR基序包含1(Sarm1)，这是Toll/IL-1受体中唯一的TIR结构域蛋白 具有NAD+水解酶活性的哺乳动物细胞。最近的研究发现了一种Sarm1细菌 CUMP直接激活同源基因是一种重要的先天免疫机制。虽然我们的研究表明 CUMP在内皮细胞感染反应中的重要作用--外酶Y是如何产生CUMP的 这导致tau的磷酸化和细胞毒性tau变体的产生，以及CUMP如何降低NAD+，同时 阻碍内皮细胞修复仍然知之甚少。为了以严格的方式解决这一知识鸿沟， 这个项目测试了铜绿假单胞菌外酶Y产生CUMP的假设，这有助于 导致肺的tau磷酸化、微管破裂和sarm1依赖的NAD+水解酶活性 损伤和阻碍修复。