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: 10626966
• 负责人: MARK D. SUTTON
• 金额: $ 77.59万
• 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/10626966
• 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; Disparate; Excision; Exposure to; Fiber; Filament; Frequencies; Future; Genetic; Genomics; Goals; Guanine; Health; Human; In Vitro; Location; Maintenance; Marketing; Medical; Mismatch Repair; Mobile Genetic Elements; Modernization; Mutagenesis; Mutation; Nucleosides; Nucleotides; 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; coping; 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：耐碳青霉烯类铜绿假单胞菌(CRPA)对人类健康构成迫切威胁 在美国和全球范围内。金属-β-内酰胺酶(MBL)，可产生高水平的碳青霉烯类耐药， 值得高度关注。CRPA产MBL假单胞菌治疗选择的匮乏 铜绿假单胞菌(MBL PA)和破裂的抗生素管道要求开发新的治疗策略 以非传统的、未开发的路径为目标。越来越多的证据表明，‘超级变种’菌株 表现出显著增加的自发突变频率(是非突变对照的10倍)，Serve 作为病理适应和抗菌素耐受的基础，不可避免地增加了治疗的可能性 PA感染中的失败和细菌持久性。重要的是，DNA复制和复制过程中的错误 跨损伤DNA合成(TLS Pol IV)是PA超突变菌株突变的机制基础。 我们已经率先合成和测试了新的非天然核苷酸，因为它非常安全和有效 抗癌疗法，这得到了我们的初步数据的支持。我们现在第一次提出要研究 通过定义超变异体在病程适应、持久性中的潜在机制来确定非天然核苷酸 和抗菌素耐药性，并开发联合方案来对抗MBL PA。我们的首要目标是 利用新型反诱变子非天然核苷酸开发治疗MBL PA的新策略 再加上现有的β-内酰胺类抗生素。治疗MBL革兰氏阴性菌的一种有希望的过渡性疗法是 头孢他啶-阿巴坦联合氨曲南；然而，这一策略在MBL PA中尚未被研究。在……里面 初步研究，我们观察到由于青霉素结合蛋白3的抑制而导致的长丝状存留。 MBL PA暴露于头孢他啶-阿维巴坦和氨曲南联合应用。因为SOS对DNA的反应 丝状化需要损伤，而TLS DNA聚合酶(POL)需要绕过DNA损伤 在导致抗菌素耐药性的周围细胞DNA中产生，我们假设在没有修复的情况下 功能，持久者应对DNA损伤并随后分离和生长的能力成为 日益依赖TLS Pol IV。鉴于PA Pol IV的这一至关重要的作用，我们的首要任务是 假设以Pol IV为靶点的新型非天然核苷酸将阻止受损DNA的复制 与现有的β-内酰胺类抗生素配合使用非常有效。为了检验这些假设，我们将：(目标1)定义 超变变子对β-内酰胺类抗生素耐药和耐受性的影响； (目的2)开发针对TLS Pol IV的小分子非天然核苷酸，以对抗MBL PA的突变； (目标3)确定非天然核苷和β-内酰胺类药物的最佳联合治疗方案 抑制耐药性，并防止MBL PA在中空纤维和动物模型中持续存在。加在一起， 我们的结果将为革兰氏阴性菌的新联合疗法提供前所未有的洞察力，并将 为未来在临床试验中测试抗突变子非天然核苷酸奠定了基础。