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: 10228661
• 负责人: BARRY Neal KREISWIRTH
• 金额: $ 81.28万
• 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 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10228661
• 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/antagonist; 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.

项目概要 脓肿分枝杆菌复合体（MABC）最近已成为脓肿病例增加的一个重要原因。 社区和医院获得性感染，特别是在免疫抑制人群中，包括 患有囊性纤维化和移植患者的人群。这种情况因其异常高的自然 并获得抗生素耐药性，使治疗复杂化，因此复杂且无效 已尝试抗生素组合，但成功率低于 50%。因此，迫切需要 改善这些感染的治疗选择。目前针对 MABC 感染的治疗建议通常是 需要单一β-内酰胺，头孢菌素、头孢西丁或碳青霉烯、亚胺培南组合 与其他药物类别，例如克拉霉素和阿米卡星。最近的研究和我们的初步结果 证明结合两种 β-内酰胺和/或 β-内酰胺酶抑制剂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（头孢他啶-亚胺培南或头孢他啶-头孢洛林） 在体外和 THP-1 细胞中对临床 MABC 具有最大的协同作用，不依赖于 β-内酰胺酶 用阿维巴坦抑制。这些结果为我们的高度发展提案提供了令人信服的科学依据 针对 MABC 感染的活性和靶向双 β-内酰胺组合。主要作为β-内酰胺类抗生素 以肽聚糖合成为目标，我们将构建新型条件肽聚糖重塑酶阻遏物 突变体来探究双β-内酰胺协同作用的分子机制，并探索有希望的 双 β-内酰胺对对抗 MABC 感染（目标 1）。因此，我们将研究各种因素之间的相互作用 肽聚糖重塑酶和一组β-内酰胺，并建立基因-化合物相互作用矩阵 双β-内酰胺效应。双β-内酰胺的转录分析将作为补充方法 揭示了对双 β-内酰胺治疗有反应的其他靶点（目标 1）。推定的β-内酰胺的功效 将针对来自多个国家的完善的 MABC 临床收集中收集的分离株对组合进行检查 美国 60 个囊性纤维化临床中心（目标 2）。赋予双β-内酰胺的自发突变体 抗性和诱导抗性突变体将进行比较基因组和 RNAseq 分析，以 确定耐药机制。 MABC 肽聚糖酶（例如 β-内酰胺酶和转肽酶） 然后将使用动力学和水解测定来探究 β-内酰胺或 β- 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) 和小鼠模型来测试 β-内酰胺的首选组合及其最佳剂量，支持 通过药代动力学 (PK) 和药效 (PD) 分析（目标 3）。在这个项目结束时，我们将 开发了针对 MABC 的新型双 β-内酰胺联合方案，并阐明了分子机制 强调协同效应的机制。此外，双-β-内酰胺对 美国各地的 MABC 分离株将被记录下来。这项研究的基因组输出将为以下研究奠定基础： 未来的工作涉及 MABC 药物开发、快速诊断和感染控制措施。知识点 关于肽聚糖酶和β-内酰胺相互作用，阻遏物和诱导突变株将是 MABC 研究界的主要资源。