Lung Endothelial Aß in infectious proteinopathy

肺内皮 A 与感染性蛋白病的关系

• 批准号: 10438793
• 负责人: Troy Stevens
• 金额: $ 38.5万
• 依托单位: UNIVERSITY OF SOUTH ALABAMA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10438793
• 关键词: Abeta synthesis; Address; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Bacteria; CRISPR/Cas technology; Cells; Characteristics; Clinical Research; Complex; Critical Illness; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cyclic GMP; Cyclic Nucleotides; Cytotoxin; Data; Death Rate; Endothelial Cells; Endothelium; Functional disorder; GTPase-Activating Proteins; Generations; Hospitals; Infection; Intensive Care Units; Knowledge; Length of Stay; Lung; Membrane; Methylation; MicroRNAs; Morbidity - disease rate; Multiprotein Complexes; Nosocomial pneumonia; Organ; Patients; Phosphorylation; Play; Prions; Process; Production; Property; Protein Kinase; Proteins; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Publishing; Reporting; Role; Severities; Specificity; Testing; Type III Secretion System Pathway; Virulence Factors; antimicrobial; beta secretase; cell injury; cytotoxic; cytotoxicity; enzyme activity; exoenzyme; gamma secretase; mortality; pathogen; presenilin-1; prevent; prion-like; protein expression; protein function; sequential proteolysis; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Pathogens responsible for nosocomial pneumonia elicit production of lung endothelial cell amyloids. These amyloids can have antimicrobial properties and be beneficial to the host, or they can have cytotoxic properties and be detrimental to the host. In the latter case, endothelial-derived amyloids may contribute to end organ dysfunction in the aftermath of critical illness. Bacterial virulence factors, such as the Pseudomonas aeruginosa type III secretion system effectors, and most notably exoenzyme Y, convert endothelial amyloids from antimicrobial to cytotoxic species. Amyloid-beta (Aβ) is one of the cytotoxic amyloids generated following endothelial infection. Aβ is released from endothelium where it becomes a transmissible and self-replicating prion cytotoxin. Our preliminary data reveals that γ-secretase activating protein plays a critical role in generating cytotoxic Aβ. We found γ-secretase activating protein expression in endothelial cells using non- biased microarray, RNAseq, methylation and miRNA screens, and confirmed protein expression and function in microvascular endothelium. γ-secretase activating protein deletion prevented formation of the cytotoxic Aβ species. Rather, following γ-secretase activating protein deletion the Pseudomonas aeruginosa- and exoenzyme Y-induced Aβ had antimicrobial properties. Exoenzyme Y is a promiscuous nucleotidyl cyclase that results in protein kinase A activation, which may phosphorylate Aβ necessary to increase its cytotoxic activity. These data suggest production of cytotoxic Aβ is due to two inter-related mechanisms, including an increase in γ-secretase activating protein function and the phosphorylation of Aβ once it is produced. Hence, this proposal tests the hypothesis that exoenzyme Y promotes the production of cytotoxic Aβ, dependent upon γ-secretase activating protein.

项目总结/摘要 导致医院获得性肺炎的病原体引起肺内皮细胞淀粉样蛋白的产生。这些 淀粉样蛋白可以具有抗微生物特性并对宿主有益，或者它们可以具有细胞毒性特性 并对宿主有害。在后一种情况下，内皮源性淀粉样蛋白可能有助于终末器官 严重疾病后的功能障碍。细菌毒力因子，如假单胞菌 铜绿III型分泌系统效应物，最显著的是外切酶Y，转化内皮淀粉样蛋白 从抗微生物到细胞毒性物种。β淀粉样蛋白（Aβ）是一种细胞毒性淀粉样蛋白， 内皮感染Aβ从内皮细胞释放，在那里它成为一种可传播和自我复制的蛋白质。 朊病毒细胞毒素我们的初步数据表明，γ-分泌酶激活蛋白在 产生细胞毒性Aβ。我们发现，在内皮细胞中γ-分泌酶激活蛋白的表达， 偏倚微阵列、RNAseq、甲基化和miRNA筛选，并证实了蛋白质表达和功能 在微血管内皮中。γ-分泌酶激活蛋白缺失可阻止细胞毒性Aβ的形成 物种相反，在γ-分泌酶激活蛋白缺失后，铜绿假单胞菌-和 外切酶Y诱导的Aβ具有抗菌活性。外切酶Y是一种混杂的核苷酸环化酶， 导致蛋白激酶A激活，这可能使Aβ磷酸化，从而增加其细胞毒性活性。 这些数据表明，细胞毒性Aβ的产生是由于两种相互关联的机制，包括增加 γ-分泌酶激活蛋白功能和Aβ一旦产生后的磷酸化。因此，本提案 检验了外切酶Y依赖于γ-分泌酶促进细胞毒性Aβ产生的假设 活化蛋白