Lung Endothelial Aß in infectious proteinopathy

肺内皮 A 与感染性蛋白病的关系

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要引起医院肺炎的病原体引起肺内皮细胞淀粉样蛋白的产生。淀粉样蛋白可以具有抗菌特性并对宿主有益，也可以具有细胞毒性在后一种情况下，内皮源性淀粉样蛋白可能对终末器官有贡献。危重疾病后的细菌毒力因子功能障碍，例如假单胞菌。铜绿假单胞菌 III 型分泌系统效应子，尤其是外酶 Y，可转化内皮淀粉样蛋白从抗菌剂到细胞毒性物质，β 淀粉样蛋白 (Aβ) 是随后产生的细胞毒性淀粉样蛋白之一。 Aβ从内皮细胞中释放出来，成为一种可传播和自我复制的物质。我们的初步数据表明，γ-分泌酶激活蛋白在朊病毒细胞毒素中发挥着关键作用。我们发现内皮细胞使用非-分泌酶激活蛋白表达。有偏差的微阵列、RNAseq、甲基化和 miRNA 筛选，并确认蛋白质表达和功能 γ-分泌酶激活蛋白的缺失阻止了细胞毒性 Aβ 的形成。相反，在 γ-分泌酶激活蛋白缺失后，铜绿假单胞菌和外酶 Y 诱导的 Aβ 具有抗菌特性。导致蛋白激酶 A 激活，这可能会使 Aβ 磷酸化，从而增加其细胞毒活性。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 γ-分泌酶激活蛋白功能和 Aβ 产生后的磷酸化因此，该建议。检验外切酶 Y 依赖于 γ 分泌酶促进细胞毒性 Aβ 产生的假设激活蛋白。