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.

项目摘要 脓肿分支杆菌复合体（MABC）最近成为导致脓肿病例增加的重要原因 社区和医院获得性感染，尤其是免疫抑制人群，包括 囊性纤维化患者和移植患者。这种情况因其异常高的自然资源而恶化。 获得性抗生素耐药性使治疗复杂化， 抗生素组合已被尝试，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 需要一种β-内酰胺，即头孢菌素、头孢西丁或碳青霉烯、亚胺培南， 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 证明了联合使用两种β-内酰胺类和/或β-内酰胺酶抑制剂可能是一种成功的策略， 治疗MABC感染。我们的研究表明，双β-内酰胺类（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的协同作用最大，不依赖于β-内酰胺酶 阿维巴坦抑制。这些结果为我们的建议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 以肽聚糖合成为靶点，我们将构建新型条件性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 针对MABC感染的双β-内酰胺对（目的1）。因此，我们将研究各种 肽聚糖重塑酶和β-内酰胺电池，并建立基因-化合物相互作用矩阵 双重β-内酰胺效应双-β-内酰胺类药物的转录分析将作为一种补充方法， 揭示了对双-β-内酰胺治疗有反应的其他靶点（目的1）。推定β-内酰胺的有效性 将针对来自多个国家的成熟MABC临床标本中采集的分离株检查组合。 美国60家囊性纤维化临床中心（目标2）。产生双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， 找出抵抗机制。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询问β-内酰胺或β-内酰胺酶的酶促机制。 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 （Hestival）和小鼠模型来测试β-内酰胺类药物的优选组合及其最佳剂量， 通过药代动力学（PK）和药效学（PD）分析（目的3）。在这个项目结束时，我们将 已经开发了针对MABC的新型双β-内酰胺联合治疗方案，并解开了MABC的分子机制。 强调协同效应的机制。此外，双-β-内酰胺类药物对 将记录美国各地的MABC分离株。这项研究的基因组输出将为以下研究奠定基础： 今后的工作涉及MABC药物开发、快速诊断和感染控制措施。知识 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被 MABC研究社区的主要资源。