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，因为它们影响药物 抵抗力和生长以及在宿主体内的存活。它包括三个目标： 具体目标1 -建立对治疗C. 沙眼感染我们将在体外测量阿奇霉素抗性（AZM*^）突变体的适合性， in vivo.然后，我们将筛选体内出现的补偿突变体，并对这些突变体进行表征 使用全基因组测序技术。我们还将选择自发性四环素耐药（Tc*^） 突变体C.在豚鼠的自然感染模型中，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 在体内和体外实验中，在没有抗生素的情况下。动物模型研究的补充将 在“治疗失败”的临床环境中进行耐药隔离调查（临床核心）。 具体目标2 -生物合成和代谢途径孔填充-表征生物化学和代谢途径孔填充。 衣原体的基本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漏洞填补联系起来， 肽聚糖悖论和转肽。我们将确定C. 沙眼衣原体对β-内酰胺类抗生素（阿莫西林/青霉素）。代谢途径的补孔成分 这一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、11 polc酸和NAD。在每种情况下，我们将研究衣原体表达的可能性， 这些因子的从头合成的酶，或者它们是否表达用于它们的 从宿主细胞质中摄取。遗传和生物化学方法将用于替代细菌 由于缺乏在衣原体中产生突变体的遗传技术， 特定目的3 -不同感染期间GPIC基因表达谱的转录组作图 豚鼠的解剖部位（眼部与生殖器）。豚鼠会感染两种不同的克隆 眼和生殖器部位的GPIC菌株和微阵列将用于确定生态位特异性基因 表达谱。待比较的表达谱为：1）眼部部位与生殖器部位; 2）生殖器部位 感染的男性与生殖器感染的女性。将选择高度差异表达的基因/途径 用于详细的体外分析（代谢），目的是探索作为靶点的途径/酶， 抑制剂和最终的新疗法。