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: 10577809
• 负责人: Peng Yuan
• 金额: $ 39.32万
• 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/10577809
• 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 Pseudomonas aeruginosa; 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; Invaded; 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; beta barrel; 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.

项目总结/摘要： 抗菌药物耐药性的上升凸显了迫切需要制定新的有效策略来对抗 抗药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， 称为伴侣引导途径（CUP）皮利，其对于病原体通过以下途径引起感染的能力至关重要： 识别和定殖不同的宿主组织和栖息地。因此，靶向组装的治疗剂 这些纤维有希望成为治疗多药耐药的急需的替代品， 耐药革兰氏阴性病原体。在这些病原体中，有一些被指定为“紧急威胁”。 碳青霉烯类耐药不动杆菌和碳青霉烯类耐药肠杆菌科（CRE），以及“严重 耐药弯曲杆菌，产超广谱β-内酰胺酶（ESBL） 肠杆菌科、多重耐药铜绿假单胞菌、耐药沙门氏菌、志贺氏菌和 百日咳杆菌在每个CUP菌毛系统中，一个指定的周质伴侣和一个外膜伴侣， (OM)引导蛋白协同工作，将数千个结构亚基组装成每个最终菌毛结构。 大多数CUP皮利也被特异性识别宿主组织中受体的粘附素所覆盖。我们取得了 在理解菌毛组装的非常复杂的机制方面取得了相当大的进展。建筑 基于我们在CUP菌毛生物发生和合理开发方面的丰富经验和专业知识， 针对CUP皮利的治疗，该提案旨在开发针对OM的新的保肢治疗 引导员使用多学科方法，包括细菌学，化学生物学，药物化学， 结构生物学和免疫学。基于这些结构特征和动态性质， 多结构域引导蛋白，我们将合理开发小分子引导抑制剂和开孔剂， 捕获特定的构象状态（Aim 1）。Usher抑制剂将解除细菌毒力因子，而 开孔剂将增加现有抗生素进入细菌外膜的渗透性。另外我们 将开发出单克隆抗体，从而阻止菌毛的生物发生和感染（Aim 2）的情况。虽然我们的前两个目标将集中在两个研究最多的菌毛系统（1型和P皮利），目标3 将扩大我们对不动杆菌属、弯曲杆菌属、铜绿假单胞菌属、沙门氏菌属、志贺氏菌属和B的研究。 百日咳。总的来说，我们计划开发针对多重耐药革兰氏阴性杆菌的合理疗法， 细菌病原体这些发展，以及我们在报告中提出的其他新战略， 多学科的U19计划，将协同工作，作为有效的药物保留疗法， 阻断引导和粘附功能。此外，在该提案中开发的usher开孔剂将 增加OM渗透性，进一步减轻革兰氏阴性病原体的抗生素耐药性， 重新利用现有的药物来增强现有的抗生素库。因此，这些领域的成功发展 方向将在对抗抗生素耐药性方面具有潜在的变革性。