Antibiotic resistance and metabolic pathways in Chlamydia species

衣原体的抗生素耐药性和代谢途径

• 批准号: 8380422
• 负责人: Anthony T Maurelli
• 金额: $ 41.32万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8380422
• 关键词: Acids; Address; Amoxicillin; Animal Model; Antibiotic Resistance; Antibiotics; Azithromycin; Biochemical; Biochemical Genetics; Cavia; Characteristics; Chlamydia; Clinical; Complement; Cytosol; Drug resistance; Enzymes; Female; Folate; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Techniques; Genital system; Goals; Growth; Growth Factor; In Vitro; Infection; Link; Maps; Measures; Metabolic; Metabolic Pathway; Modeling; Molecular; Molecular Profiling; Monobactams; Pathway interactions; Penicillins; Peptidoglycan; Reproductive Tract Infections; Resistance; Site; Surveys; System; Technology; Tetracycline Resistance; Treatment Failure; base; enzyme pathway; fitness; genome sequencing; in vivo; inhibitor/antagonist; male; mutant; novel; novel therapeutics; research study; tissue culture; uptake

Project 3 - Antibiotic resistance and metabolic pathways in Chlamydia spp This project will focus on key biosynthetic and metabolic functions of Chlamydia as they impact on drug resistance and growth and survival in the host. It consists of three aims: Specific Aim 1 - To model the emergence of resistance to the drugs of choice for the treatment of C. trachomatis infections. We will measure the fitness of azithromycin resistant (AZM*^) mutants in vitro and in vivo. We will then screen for compensatory mutants that arise in vivo and characterize these mutants using whole genome sequencing technology. We will also select for spontaneous tetracycline resistant (Tc*^) mutants of C. caviae (GPIC) in a natural infection model in guinea pigs. The mutants that arise in the natural infection setting will be analyzed for growth characteristics such as growth in tissue culture and competition experiments in vivo and in vitro in the absence of antibiotic. A complement to the animal model studies will be a survey for drug resistant Isolates the clinical setting among "treatment failures" (Clinical Core). Specific Aim 2 - Biosynthetic and metabolic pathway hole filling - characterize the biochemical and genetic components of essential biosynthetic and metabolic pathways of Chlamydia spp. This aim will link the antibiotic resistance aim of the project with biosynthetic pathway hole filling by focusing on the peptidoglycan paradox and transpeptidatlon. We will determine the molecular basis of sensitivity of C. trachomatis to p-lactam antibiotics (amoxicillin/penicillin). The metabolic pathway hole filling component of this aim will address three essential growth factors for which Chlamydia has Incomplete pathways for synthesis: folate, llpolc acid and NAD. In each case we will examine the possibility that Chlamydia express enzymes for de novo synthesis of these factors or whether they express novel transport systems for their uptake from the host cytosol. Genetic and biochemical approaches will be used in surrogate bacterial systems due to the unavailability of genetic techniques for creating mutants in Chlamydia. Specific Aim 3 - Transcriptome mapping of GPIC gene expression profile during infection in different anatomical sites (ocular vs. genital) in guinea pig. Guinea pigs will be infected with two different clonal strains of GPIC at ocular and genital sites and microarrays will be used to determine niche specific gene expression profiles. The expression profiles to be compared are: 1) ocular site vs. genital site; 2) genitally infected males vs. genitally infected females. Highly differentially expressed genes/pathways will be chosen for detailed in vitro analysis (metabdomics) with the goal of exploring pathway/enzyme as targets for inhibitors and eventually new therapeutics.

项目3-衣原体中的抗生素耐药性和代谢途径 该项目将重点放在衣原体的关键生物合成和代谢功能上，因为它们会影响药物 宿主的抵抗力，生长和生存。它由三个目标组成： 特定目的1-模拟对治疗的首选药物的抗性的出现。 沙眼感染。我们将在体外衡量阿奇霉素抗性（AZM*^）突变体的适应性和 体内。然后，我们将筛选出在体内出现并表征这些突变体的补偿性突变体 使用整个基因组测序技术。我们还将选择自发的四环素抗性（TC*^） 豚鼠自然感染模型中的C. caviae（GPIC）的突变体。自然存在的突变体 将分析感染设置的生长特征，例如组织培养和竞争的生长 在没有抗生素的情况下，体内和体外实验。对动物模型研究的补充将 对耐药性分离的调查是“治疗失败”（临床核心）之间的临床环境。 特定目标2-生物合成和代谢途径孔填充 - 表征生化和 衣原体属的必需生物合成和代谢途径的遗传成分。这个目标 将项目的抗生素抗性目的与生物合成途径孔填充联系起来。 肽聚糖悖论和肽杆菌。我们将确定C的敏感性的分子基础。 气管对P-内酰胺抗生素（阿莫西林/青霉素）。代谢途径孔填充成分 这个目标将解决衣原体为之不完整的三个基本增长因素 合成：叶酸，llpolc酸和NAD。在每种情况下，我们都会研究衣原体Express的可能性 从头综合这些因素的酶，或者它们是否表达新型运输系统 宿主细胞质的吸收。遗传和生化方法将用于替代细菌 由于无法在衣原体中创建突变体的遗传技术不可用。 特定目标3-在不同的感染过程中GPIC基因表达谱的转录组映射 豚鼠的解剖部位（眼与生殖器）。豚鼠将被两种不同的克隆感染 在眼部和生殖位点以及微阵列的GPIC菌株将用于确定特定基因 表达曲线。要比较的表达曲线是：1）眼位点与生殖器位点； 2）生殖器 被感染的雄性与受生殖感染的女性。将选择高度差异表达的基因/途径 用于详细的体外分析（替代），目的是探索途径/酶作为目标 抑制剂，最终是新的治疗剂。