Enzymology of Chlamydial Pathogenesis

衣原体发病机制的酶学

基本信息

批准号：
8697935
负责人：
Dewey G McCafferty
金额：
$ 23.53万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-03-15 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8697935
关键词：
Actins Acute Affect Animal Model Animals Apoptosis Bacteria Bacterial Infections Benchmarking Biochemical Biological Factors Biology Blindness Caspase-1 Cell Culture Techniques Cell Death Cell model Cell physiology Cells Cellular Morphology Chemicals Chlamydia Chlamydia Infections Chlamydia trachomatis Chronic Cytoskeleton Cytosol Data Defect Detection Development Disease Drug Formulations Drug Kinetics Ectopic Pregnancy Effectiveness Endothelial Cells Enzymatic Biochemistry Enzymes Epitopes Figs - dietary Genital system Goals Growth Health Hela Cells Host Defense Housing Human Immune Immunity Immunologic Markers In Vitro Infection Infection prevention Infertility Inflammation Inflammatory Intermediate Filaments Knock-out Life MEKs Mass Spectrum Analysis Mediating Methods Microtubules Modeling Molecular Motor Mus Pathogenesis Pathology Pathway interactions Pelvic Inflammatory Disease Pelvis Peptide Hydrolases Peptide Library Persons Proteins Proteolysis Proteome Publishing Recombinants Refractory Research Project Grants Role S Phase Scanning Serine Protease Serum Signal Pathway Signal Transduction Small Interfering RNA Solid Specificity Structure Trachoma Transmembrane Transport Treatment Protocols Vaccines Vacuole Virulence Factors Woman acquired immunity adaptive immunity analog antimicrobial base chemical genetics clinically relevant cytokine defense response design factor C genetic manipulation in vivo inhibitor/antagonist insight killings lipid transport men multicatalytic endopeptidase complex mutant overexpression pathogen programs public health relevance response restoration salinosporamide A scaffold small molecule tandem mass spectrometry

项目摘要

DESCRIPTION (provided by applicant): Despite aggressive antimicrobial treatment regimens, over 100 million persons are infected annually worldwide with the sexually-transmitted obligate intracellular bacterial pathogen, Chlamydia trachomatis. During infection, C. trachomatis secretes effector proteins into the host cytosol that interfere with host signaling networks to dampen host immune defenses. Unfortunately infections are asymptomatic in the majority of men and women, development of long-term protective immunity against reinfection is inhibited, and therefore many infections become chronic and spur secondary pathologies like infectious blindness (trachoma), pelvic inflammatory disorder, ectopic pregnancy, and infertility. The serine protease chlamydial protease-like activity factor (CPAF) has emerged as a central chlamydial virulence factor due to its importance in lipid and membrane transport, actin cytoskeleton structure and dynamics, microtubule-based motors, lysosomal recognition of the inclusion, ERK/MEK signaling pathways, and the onset of programmed cell death and inflammation signaling. Using an inhibitor-based chemical biology approach, we recently developed two classes of CPAF inhibitors, and observed that inhibition of CPAF activity inhibited bacterial replication, destabilized the integrity of the parasitophorous inclusion vacuole housing replicating chlamydia, and resulted in bacterial cell death. We also determined that CPAF inhibition reprogrammed infected cells to re-initiate host immune defense responses to infection by secreting pro-inflammatory cytokines and activating an inflammasome-dependent programmed host cell death pathway. Based on these collective data, we hypothesize that CPAF inhibitors may form the basis for a new class of small molecule 'antimicrobial vaccines' that not only kill bacteria, but also may enhance adaptive and acquired immunity responses to chlamydial infections. Our goals for this proposal are to understand the role CPAF in C. trachomatis pathogenesis, to gain insight into the molecular level details of CPAF protease function, inhibition and identification of targets, and to determine the extent to which CPAF inhibitors can clear infections, rescue host immune defense activity, and protect against reinfection in a clinically relevant murine animal model of chlamydial genital tract infection.

描述（由申请人提供）：尽管采取了积极的抗菌治疗方案，全世界每年仍有超过 1 亿人感染性传播的专性细胞内细菌病原体沙眼衣原体。在感染过程中，沙眼衣原体将效应蛋白分泌到宿主细胞质中，干扰宿主信号网络，抑制宿主免疫防御。不幸的是，大多数男性和女性的感染是无症状的，针对再次感染的长期保护性免疫力的发展受到抑制，因此许多感染变成慢性并引发继发性病变，如感染性失明（沙眼）、盆腔炎、宫外孕和不孕症。丝氨酸蛋白酶衣原体蛋白酶样活性因子 (CPAF) 因其在脂质和膜运输、肌动蛋白细胞骨架结构和动力学、基于微管的马达、包涵体的溶酶体识别、ERK/MEK 中的重要性而成为衣原体的中心毒力因子。信号通路，以及程序性细胞死亡和炎症信号传导的发生。使用基于抑制剂的化学生物学方法，我们最近开发了两类 CPAF 抑制剂，并观察到抑制 CPAF 活性会抑制细菌复制，破坏容纳复制衣原体的寄生包涵体液泡的完整性，并导致细菌细胞死亡。我们还确定，CPAF 抑制可重新编程受感染的细胞，通过分泌促炎细胞因子和激活炎症小体依赖性程序性宿主细胞死亡途径来重新启动宿主对感染的免疫防御反应。基于这些集体数据，我们假设 CPAF 抑制剂可能成为新型小分子“抗菌疫苗”的基础，这种疫苗不仅可以杀死细菌，还可以增强对衣原体感染的适应性和获得性免疫反应。我们本提案的目标是了解 CPAF 在沙眼衣原体发病机制中的作用，深入了解 CPAF 蛋白酶功能、抑制和靶点识别的分子水平细节，并确定 CPAF 抑制剂清除感染、挽救生命的程度。宿主免疫防御活性，并在临床相关的衣原体生殖道感染小鼠动物模型中防止再次感染。