Novel Combination Therapies to Combat Hypermutable Carbapenem-Resistant P. aeruginosa

对抗高突变碳青霉烯类耐药铜绿假单胞菌的新型联合疗法

• 批准号: 10522530
• 负责人: MARK D. SUTTON
• 金额: $ 80.51万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-24 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10522530
• 关键词: Animal Model; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Antineoplastic Agents; Attention; Aztreonam; Bacteria; Bacterial Infections; Biochemistry; Bypass; Ceftazidime; Cell division; Cells; Chemistry; Clinical Trials; Collection; Combined Antibiotics; Combined Modality Therapy; Communities; DNA; DNA Damage; DNA Repair; DNA Replication Damage; DNA biosynthesis; DNA lesion; DNA-Directed DNA Polymerase; Data; Development; Discipline; Excision; Exposure to; Fiber; Filament; Frequencies; Future; Genetic; Genomics; Goals; Guanine; Health; Human; In Vitro; Location; Maintenance; Medical; Mismatch Repair; Mobile Genetic Elements; Modernization; Monobactams; Mutagenesis; Mutation; Nucleosides; Nucleotides; Oxides; Pathway interactions; Patients; Penicillin-Binding Proteins; Pharmaceutical Preparations; Pharmacology; Phenotype; Polymerase; Process; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Public Health; Regimen; Replication Error; Research; Resistance; Role; SOS Response; Septate; System; Testing; Time; Treatment Failure; Treatment Protocols; United States; antimicrobial tolerance; arm; beta-Lactamase; beta-Lactams; cancer therapy; carbapenem resistance; combat; combinatorial; defined contribution; genome integrity; in vivo; inhibitor; insight; mortality; novel; novel therapeutic intervention; novel therapeutics; pathogen; pressure; prevent; repair function; repaired; resistance mechanism; resistant strain; small molecule; standard of care; stem; targeted agent; therapeutic target; treatment strategy; Animal Model; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Antineoplastic Agents; Attention; Aztreonam; Bacteria; Bacterial Infections; Biochemistry; Bypass; Ceftazidime; Cell division; Cells; Chemistry; Clinical Trials; Collection; Combined Antibiotics; Combined Modality Therapy; Communities; DNA; DNA Damage; DNA Repair; DNA Replication Damage; DNA biosynthesis; DNA lesion; DNA-Directed DNA Polymerase; Data; Development; Discipline; Excision; Exposure to; Fiber; Filament; Frequencies; Future; Genetic; Genomics; Goals; Guanine; Health; Human; In Vitro; Location; Maintenance; Medical; Mismatch Repair; Mobile Genetic Elements; Modernization; Monobactams; Mutagenesis; Mutation; Nucleosides; Nucleotides; Oxides; Pathway interactions; Patients; Penicillin-Binding Proteins; Pharmaceutical Preparations; Pharmacology; Phenotype; Polymerase; Process; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Public Health; Regimen; Replication Error; Research; Resistance; Role; SOS Response; Septate; System; Testing; Time; Treatment Failure; Treatment Protocols; United States; antimicrobial tolerance; arm; beta-Lactamase; beta-Lactams; cancer therapy; carbapenem resistance; combat; combinatorial; defined contribution; genome integrity; in vivo; inhibitor; insight; mortality; novel; novel therapeutic intervention; novel therapeutics; pathogen; pressure; prevent; repair function; repaired; resistance mechanism; resistant strain; small molecule; standard of care; stem; targeted agent; therapeutic target; treatment strategy

ABSTRACT: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) poses an urgent threat to human health in the United States and globally. Metallo-β-lactamases (MBL), which confer high-level carbapenem resistance, warrant significant attention. The paucity of treatment options for CRPA MBL-producing Pseudomonas aeruginosa (MBL PA) and the broken antibiotic pipeline demands the development of new therapeutic strategies that target non-traditional, unexploited pathways. There is mounting evidence that ‘hypermutator’ strains, which show a significantly increased spontaneous mutation frequency (10-fold higher than non-mutator control), serve as the basis for pathoadaptation and antimicrobial tolerance, inevitably increasing the likelihood of treatment failure and bacterial persistence in PA infections. Importantly, errors made during DNA replication and translesion DNA synthesis (TLS Pol IV) serve as the mechanistic basis for mutations in PA hypermutator strains. We have pioneered the synthesis and testing of novel non-natural nucleotides as remarkably safe and effective anti-cancer therapies, which is supported by our preliminary data. For the first time, we now propose to study non-natural nucleotides by defining the underlying mechanism of hypermutators in pathoadaptation, persistence and antimicrobial resistance and develop combination regimens to combat MBL PA. Our overarching goal is to develop new combinatorial treatment strategies for MBL PA using novel anti-mutator non-natural nucleotides together with available β-lactam antibiotics. One promising bridge therapy for MBL Gram-negatives is ceftazidime-avibactam combined with aztreonam; however, this strategy has not been studied in MBL PA. In preliminary studies, we observed long filamentous persisters due to inhibition of penicillin binding protein 3 in MBL PA exposed to the ceftazidime-avibactam and aztreonam combination. Since the SOS response to DNA damage is required for filamentation, while TLS DNA polymerases (Pols) are required to bypass DNA lesions generated in persister cell DNA leading to antimicrobial resistance, we hypothesize that in the absence of repair functions, the ability of persisters to cope with DNA damage and subsequently septate and grow becomes increasingly dependent on TLS Pol IV. Given this critically important role of PA Pol IV, our overarching hypothesis that novel, non-natural nucleotides that target Pol IV to block replication of damaged DNA will be highly effective together with existing β-lactam antibiotics. To test these hypotheses, we will: (Aim 1) define the contributions of hypermutators to resistance and persistence of MBL PA exposed to β-lactam combinations; (Aim 2) develop small molecule, non-natural nucleotides targeting TLS Pol IV to combat mutation in MBL PA; (Aim 3) define optimal combinatorial treatment regimens of non-natural nucleosides and β-lactams that suppresses resistance, and prevents persistence of MBL PA in hollow fiber and animal models. Taken together, our results will provide unprecedented insight into novel combination therapies for Gram-negatives, and will set the cornerstone for future testing of anti-mutator non-natural nucleotides in clinical trials.

摘要：抗碳青霉烯铜绿假单胞菌（CRPA）对人类健康有紧迫的威胁 在美国和全球。金属-β-内乳酶（MBL），会议高级碳青霉烯耐药性， 值得关注。 CRPA MBL产生假单胞菌的治疗选择匮乏 铜绿（MBL PA）和破裂的抗生素管道需要开发新的治疗策略 该目标是非传统的，意外的途径。有越来越多的证据表明“高压器”菌株， 显示自发突变频率明显增加（比非杂物控制高10倍），服务 作为疗法和抗菌耐受性的基础，不可避免地会增加治疗的可能性 PA感染的衰竭和细菌持久性。重要的是，在DNA复制期间犯了错误 Translesion DNA合成（TLS POLI IV）是PA超敏化菌株突变的机械基础。 我们开创了新型非天然核动肽的合成和测试是非常安全有效的 抗癌疗法，由我们的初步数据支持。我们现在第一次提议学习 非天然核苷酸通过定义途径持续性的高压剂的潜在机制 以及抗菌素的耐药性和发育组合方案，以对抗MBL PA。我们的总体目标是 使用新型抗突变器非天然核苷酸为MBL PA制定新的组合治疗策略 以及可用的β-内酰胺抗生素。一种有望用于MBL革兰氏阴性剂的桥梁疗法是 Ceftazidime-Avibactam与Aztreonam结合；但是，该策略尚未在MBL PA中研究。在 初步研究，我们观察到由于青霉素结合蛋白3的抑制，我们观察到了长丝状持久。 MBL PA暴露于Ceftazidime-Avibactam和Aztreonam组合。由于SOS对DNA的响应 细丝需要损坏，而TLS DNA聚合酶（POLS）绕过DNA病变需要 在持久的细胞DNA中产生，导致抗菌素耐药性，我们假设在没有修复的情况下 功能，持续应对DNA损伤的能力以及随后分隔和生长的能力变成 越来越多地依赖于TLS pol IV。鉴于Pa Pol IV的这一至关重要的作用，我们的总体 假设靶向pol IV以阻断受损DNA复制的新型非天然核苷酸将是 与现有的β-内酰胺抗生素一起高效。为了检验这些假设，我们将：（目标1）定义 高压器对暴露于β-内酰胺组合的MBL PA的耐药性和持久性的贡献； （AIM 2）开发针对TLS Pol IV的小分子，非天然核动肽以对抗MBL PA中的突变； （目标3）定义非天然核苷和β-内酰胺的最佳组合治疗方案 抑制阻力，并防止在空心纤维和动物模型中MBL PA的持久性。在一起， 我们的结果将为革兰氏阴性剂的新型组合疗法提供前所未有的见解，并将设定 在临床试验中，抗突袭非天然核苷酸的未来测试的基石。