A dual-beta-lactam strategy for treating multidrug resistant M abscessus

治疗多重耐药脓肿分枝杆菌的双 β-内酰胺策略

• 批准号: 10457876
• 负责人: BARRY Neal KREISWIRTH
• 金额: $ 80.99万
• 依托单位: HACKENSACK UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10457876
• 关键词: Amikacin; Aminoglycosides; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antimycobacterial Agents; Binding; Biological Assay; Carbapenems; Cefoxitin; Ceftazidime; Cells; Cephalosporins; Clarithromycin; Clinical; Collection; Combined Antibiotics; Communities; Community Hospitals; Complex; Cystic Fibrosis; Data; Dose; Drug Targeting; Drug resistance; Effectiveness; Enzyme Interaction; Enzymes; Evaluation; Exposure to; Fiber; Future; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Goals; Growth; Hydrolysis; Imipenem; In Vitro; Infection; Infection Control; Kinetics; Knowledge; Lung infections; Macrolide-resistance; Macrolides; Mass Spectrum Analysis; Measures; Modeling; Molecular; Monobactams; Multi-Drug Resistance; Mus; Mycobacterium abscessus; Nosocomial Infections; Output; Peptidoglycan; Peptidyltransferase; Pharmaceutical Preparations; Population; Rapid diagnostics; Recommendation; Regimen; Research; Resistance; Resources; Signal Transduction; Suggestion; Testing; Therapeutic; Transplant Recipients; Treatment Protocols; Work; base; beta-Lactam Resistance; beta-Lactamase; beta-Lactams; chronic infection; clinical center; clinically relevant; comparative genomics; conditional mutant; drug development; effective therapy; gene repression; immunosuppressed; improved; inhibitor; mouse model; multi-drug resistant pathogen; mutant; novel; novel therapeutic intervention; pharmacokinetics and pharmacodynamics; resistance mechanism; response; screening; success; synergism; transcriptome sequencing

PROJECT SUMMARY Mycobacterium abscessus complex (MABC) has recently emerged as a significant cause of increasing cases of both community- and hospital-acquired infections, especially among immunosuppressed populations, including populations with cystic fibrosis and transplant patients. This situation is worsened by its exceptionally high natural and acquired antibiotic resistance that complicates treatment, and consequently, complex and ineffective antibiotic combinations have been tried with success rates below 50%. As a result, there is an urgent need to improve therapeutic options for these infections. Current treatment recommendations for MABC infection usually requires a single β-lactam, either the cephalosporin, cefoxitin, or the carbapenem, imipenem, to be combined with other drug classes, e.g. clarithromycin and amikacin. Recent studies and our preliminary results demonstrated that combining two β-lactams and/or a β-lactamase inhibitor could be a successful strategy to treat MABC infections. Our studies showed dual-β-lactams (ceftazidime-imipenem or ceftazidime-ceftaroline) had the greatest synergic effects against clinical MABC in vitro and in THP-1 cells, independent of β-lactamase inhibition with avibactam. These results provide a compelling scientific basis for our proposal to develop highly active and targeted dual-β-lactam combinations against MABC infections. As β-lactam antibiotics primarily target peptidoglycan synthesis, we will construct novel conditional peptidoglycan remodeling enzyme repressor mutants to interrogate the molecular mechanisms underlying dual-β-lactam synergy, and to probe promising dual-β-lactam pairs against MABC infections (Aim 1). As such, we will examine the interactions between various peptidoglycan remodeling enzymes and a battery of β-lactams, and build up a gene-compound interaction matrix of dual-β-lactam effects. Transcriptional analysis of dual-β-lactams will be used as a complimentary approach to reveal additional targets responsive to dual-β-lactam treatment (Aim 1). The efficacy of putative β-lactam combinations will be examined against isolates collected in a well-established MABC clinical collection from over 60 cystic fibrosis clinical centers across the US (Aim 2). Spontaneous mutants conferring dual-β-lactam resistance and induced resistance mutants will be subjected to comparative genomic and RNAseq analysis to identify the resistance mechanisms. MABC peptidoglycan enzyme (e.g. β-lactamase and transpeptidases) kinetic and hydrolysis assays will then be used to interrogate the enzymatic mechanism of β-lactams or a β- lactamase inhibitor against MABC infections. Lastly, we will use the state-of-art hollow fiber infection model (HFIM) and mouse models to test the preferred combinations of β-lactams and their optimal doses, supported by pharmacokinetic (PK) and pharmacodynamic (PD) analyses (Aim 3). At the conclusion of this project, we will have developed novel dual-β-lactam combination regimens against MABC, and unraveled the molecular mechanism underlining the synergistic effects. In addition, the effectiveness the dual-β-lactams against the MABC isolates across the U.S. will be documented. The genomic output of this study will serve the basis for future work on MABC drug development, rapid diagnostics and infection control measures. The knowledge regarding peptidoglycan enzyme and β-lactam interaction, and the repressor and induced mutant strains will be a major resource for the MABC research community.

项目总结 脓肿分枝杆菌复合体(MABC)最近已成为增加的病例的一个重要原因。 社区和医院获得性感染，特别是在免疫抑制人群中，包括 囊性纤维化症患者和移植患者。这种情况因其异常高的自然环境而变得更加糟糕 并获得抗生素耐药性，使治疗复杂化，因此，复杂和无效 抗生素组合已经尝试过，成功率低于50%。因此，迫切需要 改善这些感染的治疗选择。目前对MABC感染的治疗建议通常 需要单一的β-内酰胺类药物，头孢菌素、头孢西丁或碳青霉烯、亚胺培南联合使用 与其他药物类别，如克拉霉素和阿米卡星。最近的研究和我们的初步结果 证明了将两种β-内酰胺类药物和/或一种β-内酰胺酶抑制剂联合使用可能是一种成功的策略 治疗MABC感染。我们的研究显示双β-内酰胺类(头孢他啶-亚胺培南或头孢他啶-头孢他林) 在体外和在THP-1细胞中对临床MABC有最大的协同作用，不依赖β-内酰胺酶 阿维巴坦的抑制作用。这些结果为我们的提议提供了令人信服的科学基础，使其高度 积极的和有针对性的双β-内酰胺组合，以对抗MABC感染。以β-内酰胺类抗生素为主 以肽聚糖的合成为目标，构建新的条件性肽聚糖重塑酶抑制因子 突变体询问双β-内酰胺协同作用的分子机制，并探索有希望的 抗MABC感染的双β-内酰胺对(目标1)。因此，我们将研究不同类型的 肽聚糖重塑酶和一组β-内酰胺类药物，并建立基因-化合物相互作用矩阵 双β-内酰胺类药物的作用。双β-内酰胺类药物的转录分析将作为补充方法用于 揭示对双β-内酰胺治疗有反应的其他目标(目标1)。推定的β-内酰胺类药物的疗效 组合将与从来自OVER的已建立的MABC临床收集中收集的分离株进行对照检验 全美60个囊性纤维化临床中心(AIM 2)。产生双β-内酰胺的自发突变株 抗性和诱导抗性突变体将接受比较基因组和RNAseq分析 找出抵抗机制。单链多糖酶(如β-内酰胺酶和转肽酶) 然后，将使用动力学和水解分析来询问β-内酰胺类或β-内酰胺类的酶机制。 抗MABC感染的内酰胺酶抑制剂。最后，我们将使用最先进的中空纤维感染模型 (HFIM)和小鼠模型，以测试β-内酰胺类药物的首选组合及其最佳剂量，支持 通过药代动力学(PK)和药效学(PD)分析(目标3)。在这个项目结束时，我们将 开发了新的抗MABC的双β-内酰胺联合方案，并解开了分子 强调协同效应的机制。此外，双β-内酰胺类药物对沙门氏菌病的疗效 美国各地的MABC分离株将被记录在案。这项研究的基因组结果将为 未来在MABC药物开发、快速诊断和感染控制措施方面的工作。《知识》 关于肽聚糖酶和β-内酰胺的相互作用，以及阻遏和诱导突变菌株将 MABC研究界的主要资源。