Antibiotic-sparing strategies targeting outer membrane ushers in Gram-negative bacterial pathogens

针对外膜的抗生素节约策略迎来革兰氏阴性细菌病原体

• 批准号: 10352470
• 负责人: Peng Yuan
• 金额: $ 36.09万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10352470
• 关键词: Acinetobacter; Address; Adhesions; Age; Antibiotic Resistance; Antibiotics; Antibodies; Bacterial Adhesins; Bacterial Drug Resistance; Bacterial Infections; Bacteriology; Binding; Biochemical; Biogenesis; Biological Assay; Biology; Bordetella pertussis; Campylobacter; Cell Membrane Permeability; Centers for Disease Control and Prevention (U.S.); Chemicals; Communities; Complex; Development; Diagnosis; Drug resistance; Drug-resistant Campylobacter; Enterobacteriaceae; Escherichia coli; Extended-spectrum β-lactamase; Fiber; Fimbriae Proteins; Funding Opportunities; Gram-Negative Bacteria; Habitats; Immunoglobulins; Immunology; In Vitro; Individual; Infection; International; Knowledge; Ligand Binding Domain; Mediating; Membrane; Microbial Biofilms; Molecular; Molecular Chaperones; Molecular Conformation; Monoclonal Antibodies; Multi-Drug Resistance; Nature; Pathway interactions; Permeability; Persons; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pilum; Predisposition; Prevention therapy; Pseudomonas aeruginosa; Public Health; Reporting; Role; Salmonella; Shapes; Shigella; Structure; Surface; System; Tertiary Protein Structure; Therapeutic; Therapeutic Monoclonal Antibodies; Tissues; United States; Urinary tract infection; Uropathogenic E. coli; Usher Proteins; Virulence; Virulence Factors; Work; alternative treatment; antibiotic resistant infections; appendage; base; carbapenem resistance; carbapenem-resistant Enterobacteriaceae; combat; drug resistant pathogen; efficacy testing; experience; extracellular; in vivo; inhibitor; innovation; interdisciplinary approach; multidisciplinary; multidrug-resistant Pseudomonas aeruginosa; novel; novel strategies; pathogen; pathogenic bacteria; periplasm; prevent; programs; receptor; recruit; resistant Shigella; small molecule; small molecule libraries; structural biology; targeted treatment; therapeutic development; therapeutic target; therapy development; treatment strategy; Acinetobacter; Address; Adhesions; Age; Antibiotic Resistance; Antibiotics; Antibodies; Bacterial Adhesins; Bacterial Drug Resistance; Bacterial Infections; Bacteriology; Binding; Biochemical; Biogenesis; Biological Assay; Biology; Bordetella pertussis; Campylobacter; Cell Membrane Permeability; Centers for Disease Control and Prevention (U.S.); Chemicals; Communities; Complex; Development; Diagnosis; Drug resistance; Drug-resistant Campylobacter; Enterobacteriaceae; Escherichia coli; Extended-spectrum β-lactamase; Fiber; Fimbriae Proteins; Funding Opportunities; Gram-Negative Bacteria; Habitats; Immunoglobulins; Immunology; In Vitro; Individual; Infection; International; Knowledge; Ligand Binding Domain; Mediating; Membrane; Microbial Biofilms; Molecular; Molecular Chaperones; Molecular Conformation; Monoclonal Antibodies; Multi-Drug Resistance; Nature; Pathway interactions; Permeability; Persons; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pilum; Predisposition; Prevention therapy; Pseudomonas aeruginosa; Public Health; Reporting; Role; Salmonella; Shapes; Shigella; Structure; Surface; System; Tertiary Protein Structure; Therapeutic; Therapeutic Monoclonal Antibodies; Tissues; United States; Urinary tract infection; Uropathogenic E. coli; Usher Proteins; Virulence; Virulence Factors; Work; alternative treatment; antibiotic resistant infections; appendage; base; carbapenem resistance; carbapenem-resistant Enterobacteriaceae; combat; drug resistant pathogen; efficacy testing; experience; extracellular; in vivo; inhibitor; innovation; interdisciplinary approach; multidisciplinary; multidrug-resistant Pseudomonas aeruginosa; novel; novel strategies; pathogen; pathogenic bacteria; periplasm; prevent; programs; receptor; recruit; resistant Shigella; small molecule; small molecule libraries; structural biology; targeted treatment; therapeutic development; therapeutic target; therapy development; treatment strategy

PROJECT SUMMARY/ ABSTRACT: The rise of antibacterial resistance highlights the urgent need to develop new effective strategies to combat antibiotic-resistant infections. Ubiquitously, Gram-negative bacterial pathogens assemble extracellular fibers, termed chaperone-usher pathway (CUP) pili, that are critical for the pathogen's ability to cause infections by recognizing and colonizing different host tissues and habitats. Thus, therapeutics targeting the assembly of these fibers hold promise in their potential to result in much needed alternatives for the treatment of multidrug- resistant Gram-negative pathogens. Among these pathogens are those designated as “Urgent Threats” carbapenem-resistant Acinetobacter and carbapenem-resistant Enterobacteriaceae (CRE), as well as “Serious Threats” drug-resistant Campylobacter, extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae, multidrug-resistant Pseudomonas aeruginosa, drug-resistant Salmonella, Shigella, and Bordetella pertussis. In each CUP pilus system, a designated periplasmic chaperone and an outer-membrane (OM) usher protein work together to assemble thousands of structural subunits into each final pilus structure. Most CUP pili are also tipped by adhesins that specifically recognize receptors in host tissues. We have made considerable progress towards understanding the remarkably complex mechanisms of pilus assembly. Building on our extensive experience and expertise in CUP pilus biogenesis and in the development of rational therapies targeting CUP pili, this proposal seeks to develop novel antibiotic-sparing therapies targeting the OM ushers using multidisciplinary approaches including bacteriology, chemical biology, medicinal chemistry, structural biology and immunology. Based on the structural characterizations and the dynamic nature of these multi-domain usher proteins, we will rationally develop small molecule usher inhibitors and pore openers by trapping specific conformational states (Aim 1). Usher inhibitors will disarm bacterial virulence factors, whereas pore openers will increase permeability of existing antibiotics into bacterial outer membranes. In addition, we will develop monoclonal antibodies that inactivate usher, thus preventing pilus biogenesis and infection (Aim 2). While our first two aims will concentrate on two of the most studied pilus systems (type 1 and P pili), Aim 3 will expand our studies of ushers in Acinetobacter, Campylobacter, P. aeruginosa, Salmonella, Shigella, and B. pertussis. Collectively, we plan to develop rational therapies against multiple antibiotic-resistant Gram-negative bacterial pathogens. These developments, together with other novel strategies proposed in our multidisciplinary U19 program, will work synergistically to act as efficient antibiotic-sparing therapeutics by blocking usher and adhesin functions. Moreover, the usher pore openers developed in this proposal will increase OM permeability, further alleviating antibiotic resistance in Gram-negative pathogens and allowing us to repurpose existing drugs to enhance the current antibiotic arsenal. Thus, successful developments in these directions will be potentially transformative in combating antibiotic resistance.

项目摘要/摘要： 抗菌抗性的兴起凸显了迫切需要制定新有效策略来对抗 抗生素抗性感染。普遍存在的革兰氏阴性细菌病原体组装细胞外纤维， 称为伴侣 - 垫圈途径（杯）pili，这对于病原体引起感染的能力至关重要 识别和定居不同的宿主组织和栖息地。那是针对组装的治疗 这些纤维具有潜力，有望为治疗多饮用的急需替代方案 抗革兰氏阴性病原体。这些病原体中有那些被指定为“紧急威胁”的病原体 抗碳青霉烯菌和碳青霉烯二杆菌科（CRE）以及“严重的 威胁”耐药的弯曲杆菌，扩展光谱β-内酰胺酶（ESBL） - 生产 肠杆菌科，抗多药的铜绿假单胞菌，耐药沙门氏菌，志贺氏菌和 Bordetella百日咳。在每个杯子活塞系统中，指定的周质伴侣和外膜 （OM）USHER蛋白共同起作用，将成千上万个结构亚基组装到每个最终的Pyrus结构中。 大多数杯子菌毛也被特异性识别宿主组织中的受体的粘附器倾斜。我们做了 朝着理解pyrus组装的复杂机制的巨大进展。建筑 关于我们在杯状生物发生方面的丰富经验和专业知识和理性发展 该提案旨在针对杯子pili的疗法，旨在开发针对OM的新型抗生素疗法 使用多学科方法的引诱者，包括细菌，化学生物学，药物化学， 结构生物学和免疫学。基于结构特征和这些的动态性质 多域usher蛋白，我们将通过合理地发展小分子usher抑制剂和孔隙开启器 捕获特定的会议状态（目标1）。 usher抑制剂会解除细菌病毒因素，而 开口机将增加现有抗生素对细菌外膜的渗透性。另外，我们 将开发单克隆抗体，从而使usher失活，从而防止活塞生物发生和感染（AIM 2）。虽然我们的前两个目标将集中在两个研究最多的活塞系统（1型和P Pilli）上，AIM 3 将扩大我们对流动杆菌，弯曲杆菌，铜绿假单胞菌，沙门氏菌，志贺氏菌和B的研究的研究。 百日咳。总的来说，我们计划开发针对多种抗抗生素革兰氏阴性剂的合理疗法 细菌病原体。这些事态发展以及我们在我们的其他新颖策略一起 多学科U19计划将协同起作用，以通过 阻止Usher和粘合功能。此外，本提案中开发的迎风孔开瓶器将 增加OM渗透性，进一步减轻革兰氏阴性病原体中的抗生素耐药性并允许我们 重新利用现有药物以增强当前的抗生素武器库。这是这些成功的发展 方向将在对抗抗生素耐药性方面具有潜在的变化。