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 最近出现的腹肌分枝杆菌（MABC）已成为增加病例的重要原因 社区和医院获得的感染，尤其是在免疫抑制人群中，包括 囊性纤维化和移植患者的种群。这种情况由于其异常高的自然而恶化了 并获得了使治疗复杂化的抗生素耐药性，因此，复杂而无效 抗生素组合已尝试，成功率低于50％。结果，迫切需要 改善这些感染的治疗选择。通常对MABC感染的治疗建议 需要单个β-内酰胺，即头孢菌素，头孢辛蛋白或碳青霉烯，imipenem 与其他药物类别，例如克拉霉素和amikacin。最近的研究和我们的初步结果 证明将两个β-内酰胺和/或β-内酰胺酶抑制剂结合在一起可能是成功的策略 治疗MABC感染。我们的研究表明双β-内酰胺（copazidime-Imipemenem或Ceftazidime-cefttaroline） 在体外和THP-1细胞中对临床MABC具有最大的协同作用，与β-内酰胺酶无关 用Avibactam抑制。这些结果为我们的建议提供了令人信服的科学基础 针对MABC感染的活性和靶向双β-内酰胺组合。作为β-内酰胺抗生素主要 靶肽聚糖合成，我们将构建新的有条件肽聚糖重塑酶复制品 突变体询问双β-内酰胺协同作用的分子机制，并探测有希望的 双β-内酰胺与MABC感染配对（AIM 1）。因此，我们将研究各种 肽聚糖重塑酶和一组β-内酰胺，并构建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用作一种免费的方法 揭示对双β-内酰胺治疗有效的其他靶标（AIM 1）。推定的β-内酰胺的效率 将检查组合，以针对从越来越公认的MABC临床收集中收集的分离株进行检查 60个囊性纤维化临床中心遍布整个美国（AIM 2）。自发突变体会议双β-内酰胺 抗性和诱导的抗性突变体将经过比较基因组和RNASEQ分析对 确定电阻机制。 MABC Pepperidoglycan酶（例如β-内酰胺酶和转肽酶） 然后，动力学和水解测定法将用于询问β-内酰胺或β-的酶促机理 针对MABC感染的乳糖酶抑制剂。最后，我们将使用最先进的空心纤维感染模型 （HFIM）和小鼠模型，以测试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）和药效学（PD）分析（AIM 3）。在这个项目的结论下，我们将 已经开发了针对MABC的新型双β-内酰胺组合方案，并揭示了分子 强调协同作用的机制。另外，有效性双β-内酰胺针对 将记录美国MABC分离株。这项研究的基因组输出将为 MABC药物开发，快速诊断和感染控制措施的未来工作。知识 有关肽聚糖酶和β-内酰胺的相互作用，复制剂和诱导的突变菌株将是 MABC研究社区的主要资源。