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: 7762442
• 负责人: Anthony T Maurelli
• 金额: $ 38.13万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-21 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7762442
• 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 - 衣原体的抗生素耐药性和代谢途径 该项目将重点关注衣原体的关键生物合成和代谢功能，因为它们对药物的影响 宿主的抵抗力、生长和存活。它由三个目标组成： 具体目标 1 - 模拟对治疗艰难梭菌所选药物的耐药性的出现。 沙眼衣原体感染。我们将在体外测量阿奇霉素抗性（AZM*^）突变体的适合度，并 体内。然后我们将筛选体内出现的补偿突变体并表征这些突变体 使用全基因组测序技术。我们还将选择自发四环素耐药性 (Tc*^) 豚鼠自然感染模型中的豚鼠线虫（GPIC）突变体。自然界中出现的突变体 将分析感染环境的生长特征，例如组织培养中的生长和竞争 在没有抗生素的情况下进行体内和体外实验。动物模型研究的补充将 进行耐药性调查 隔离“治疗失败”的临床环境（临床核心）。 具体目标 2 - 生物合成和代谢途径孔洞填充 - 表征生化和 衣原体必需生物合成和代谢途径的遗传成分。这一目标将 将项目的抗生素耐药性目标与生物合成途径孔洞填充联系起来，重点关注 肽聚糖悖论和转肽。我们将确定 C 敏感性的分子基础。 沙眼衣原体转用 p-内酰胺类抗生素（阿莫西林/青霉素）。代谢途径孔洞填充成分 该目标将解决衣原体具有不完整途径的三个基本生长因子 合成：叶酸、llpolc酸和NAD。在每种情况下，我们都会检查衣原体表达的可能性 用于从头合成这些因子的酶，或者它们是否表达新的转运系统 从宿主细胞质中摄取。遗传和生化方法将用于替代细菌 由于无法在衣原体中产生突变体的遗传技术，系统的发展受到了限制。 具体目标 3 - 不同感染过程中 GPIC 基因表达谱的转录组图谱 豚鼠的解剖部位（眼部与生殖器）。豚鼠将感染两种不同的克隆 眼部和生殖器部位的 GPIC 菌株和微阵列将用于确定生态位特异性基因 表达谱。要比较的表达谱是：1) 眼部位与生殖部位； 2) 生殖器方面 受感染的男性与生殖器受感染的女性。将选择高度差异表达的基因/途径 用于详细的体外分析（元组学），目的是探索途径/酶作为目标 抑制剂和最终的新疗法。