Defining human interferon-stimulated genes with novel functions in host defense to Chlamydia infections

定义在宿主防御衣原体感染方面具有新功能的人干扰素刺激基因

• 批准号: 9911637
• 负责人: Stephen Charles Walsh
• 金额: $ 3.75万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9911637
• 关键词: Acids; Anti-Bacterial Agents; Antibacterial Response; Bacteria; Biological; Biological Assay; Biology; Cells; Chlamydia; Chlamydia Infections; Chlamydia muridarum; Chlamydia trachomatis; Coenzyme A; Detection; Development; Diagnostic; Disease; Ectopic Pregnancy; Environment; Enzymes; Epidemic; Epithelial Cells; Genes; Genetic; Genetic Screening; Genomics; Growth; Host Defense; Human; Hypersensitivity; Immune; Immune response; Immune system; Immunity; Infection; Infertility; Interferon Type II; Interferons; Interleukins; Invaded; Lead; Life; Ligase; Lipids; Mediating; Medical; Medicine; Microbe; Microscopic; Molecular; Mus; Natural Immunity; Nature; Pathogenesis; Pathogenicity; Pelvic Inflammatory Disease; Pharmaceutical Preparations; Phenotype; Proteins; Resistance; Role; Sexual Transmission; Single Nucleotide Polymorphism; Site; Techniques; Testing; Therapeutic; Vaccines; Virulence; Virulence Factors; Woman; Work; antimicrobial; bacterial genetics; base; causal variant; chronic infection; cytokine; experimental study; functional genomics; gene function; genetic element; genetic variant; genome sequencing; immune clearance; improved; long chain fatty acid; medical complication; microbial; mutant; mycobacterial; novel; pathogen; pathogenic bacteria; pathogenic microbe; programs; prophylactic; resistance mechanism; response; small molecule libraries; trait; vaccination strategy; whole genome

ABSTRACT Chlamydia trachomatis is the most widespread sexually transmitted bacterial pathogen in the world. People infected with C. trachomatis are often initially asymptomatic, hindering the proper diagnostic and therapeutic strategies necessary to impede this “silent epidemic”. If not properly treated, the bacteria are able to establish a long-lasting, persistent infection that can ultimately lead to severe medical sequelae. These complications arise predominantly in women, and include pelvic inflammatory disease, life-threatening ectopic pregnancies or infertility. A critical component of the microbial pathogenesis of C. trachomatis is its ability to evade immune detection and other antimicrobial responses conferred by its human host. These host defenses are largely galvanized by the cytokine interferon-gamma (IFNγ), which stimulates epithelial cells occupying the site of an infection to up regulate the expression of interferon-stimulated-genes (ISGs). These ISGs are then responsible for the execution and immune clearance of invading microbes. In order to subvert the effects of IFNγ and successfully replicate in epithelial cells, C. trachomatis must have evolved counterdefenses to ISGs that normally target and destroy other bacterial pathogens. However, the identity and function of these ISGs, as well as the C. trachomatis virulence effectors that inhibit ISG functions, are poorly understood. In pursuit of answering these questions, we performed two complementary screens to identify i) ISGs with anti-Chlamydia activities and ii) C. trachomatis genetic mutants with hypersensitivity to IFNγ treatment. In Aim 1 of this proposal, we will use a combination of functional genetics and cell biological studies in human cells to dissect the intracellular responses conferred by these anti-Chlamydia ISGs. In Aim 2, we will combine parallel approaches in bacterial genetics and whole-genome sequencing to pinpoint the causative genetic elements responsible for C. trachomatis evasion of IFNγ-mediated immunity. Taken together, these experiments will interrogate the dynamic relationship between cell-intrinsic defenses mediated by human ISGs and counter- resistance mechanisms of Chlamydia that are employed during infection. Implications of these studies will provide important platforms for the development of novel anti-Chlamydia medicines or vaccination strategies that treat its associated disease.

抽象的 沙眼衣原体是世界上最广泛的性传播细菌病原体。 感染沙眼炎的人通常是不对称的，阻碍了适当的诊断和 阻碍这种“无声流行”所必需的治疗策略。如果没有适当的治疗，细菌可以 建立持久的持续感染，最终会导致严重的医学后遗症。这些 并发症主要在女性中出现，包括骨盆炎症性疾病，威胁生命的生态 怀孕或不育。沙眼梭状芽胞庭的微生物发病机理的关键成分是它的能力 逃避免疫检测和其他由人类宿主提供的抗菌反应。这些主机防御 细胞因子干扰素γ（IFNγ）在很大程度上镀锌，刺激上皮细胞占据 感染部位上调干扰素刺激的基因（ISGS）的表达。这些ISG是 负责入侵微生物的执行和免疫清除。为了颠覆 IFNγ并在上皮细胞中成功复制，沙眼梭状芽孢杆菌必须使ISGS进化为反防御 这通常靶向并破坏其他细菌病原体。但是，这些ISG的身份和功能， 以及抑制ISG功能的沙眼病毒效应以及众所周知。追求 回答这些问题，我们执行了两个完整的屏幕来识别I）ISGS与抗辣椒的屏幕 活性和ii）对IFNγ治疗过敏的气管梭菌基因突变体。在目标1中 提案，我们将在人类细胞中使用功能遗传学和细胞生物学研究的组合来剖析 这些抗智慧ISG赋予的细胞内反应。在AIM 2中，我们将合并平行 细菌遗传学和全基因组测序的方法，以查明原因 负责IFNγ介导的免疫力的沙眼曲霉演变。两者一起，这些实验将 询问人类ISG介导的细胞中性防御与反相反的动态关系 在感染过程中采用衣原体的抗性机制。这些研究的含义将 为开发新型抗智慧药物或疫苗接种策略提供重要平台 治疗其相关疾病。