Developing protective monoclonal antibodies against Gram- and Gram+ bacteria

开发针对革兰氏菌和革兰氏菌的保护性单克隆抗体

• 批准号: 10577803
• 负责人: Ali Hassan Ellebedy
• 金额: $ 38.6万
• 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/10577803
• 关键词: Acinetobacter; Adherence; Adhesives; Antibiotic Resistance; Antibiotics; Bacteria; Bacterial Adhesins; Bacterial Drug Resistance; Binding; Bladder; Bordetella pertussis; Catheters; Cells; Centers for Disease Control and Prevention (U.S.); Clinical; Cloning; Collaborations; Combating Antibiotic Resistant Bacteria; Communicable Diseases; Contracts; Development; Drug resistance; Drug resistant Pseudomonas aeruginosa; Drug-resistant Campylobacter; Drug-resistant Neisseria Gonorrhoeae; Enterobacteriaceae; Enterococcus; Enzymes; Epitopes; Excision; Extended-spectrum β-lactamase; Fiber; Fibrinogen; Future; Gastrointestinal tract structure; Generations; Goals; Gram-Negative Bacteria; Health; Host Defense Mechanism; Human; Indwelling Catheter; Infection; Interdisciplinary Study; Invaded; Kidney; Klebsiella; Mannose; Mediating; Membrane; Methods; Microbial Biofilms; Molecular Chaperones; Molecular Machines; Monoclonal Antibodies; Monoclonal Antibody Therapy; Mucous Membrane; Multi-Drug Resistance; Mutation; Nature; Operon; Pathway interactions; Persons; Pilum; Polymers; Proteins; Public Health; Pump; Reporting; Resistance; Resistance development; Salmonella; Serotyping; Site; Surface; System; Techniques; Testing; Therapeutic; Therapeutic Monoclonal Antibodies; Time; Tissues; Transmembrane Domain; United States; Uropathogenic E. coli; Usher Proteins; Vaccine Design; Virulence; Work; alternative treatment; antibiotic resistant infections; carbapenem resistance; carbapenem-resistant Enterobacteriaceae; combat; drug resistant pathogen; extracellular; fluid flow; gene conservation; host colonization; human tissue; innovation; multi-drug resistant pathogen; multidrug-resistant Pseudomonas aeruginosa; non-typhoidal Salmonella; pathogen; pathogenic bacteria; periplasm; pilus genes; polymerization; pressure; prevent; programs; residence; resistance mechanism; resistant Shigella; sortase; success; therapeutic development; tool; urinary

PROJECT SUMMARY/ ABSTRACT SUMMARY: Antibiotic resistance is increasingly a threat to human health resulting in difficult to treat infections, with some multidrug resistant pathogens being resistant to all or most all antibiotics. To combat this growing crisis, this U19 is taking an integrated approach to the generation and development of therapeutics against key virulence mechanism of bacteria. Specifically, this CORE will in an integrated approach with Projects 1 and 2, seek to develop monoclonal antibody (mAb) therapies against different bacterial adhesins that extensive studies have shown to be critical for the ability of pathogens to bind to and resist removal from host tissues and surfaces. Uropathogenic E. coli (UPEC) use CUP adhesive pili tipped with adhesin proteins to bind to a variety of human tissues including the uninflammed bladder, the inflamed bladder, the kidney, urinary catheters and the gastrointestinal tract (GIT). In addition, Klebsiella expresses a mannose binding FimH adhesin that is critical for the ability of Klebsiella to cause UTIs. Further, CUP adhesins in Acinetobacter baumanii, CupD and PrpD, as well as the EbpA adhesin at the tip of the sortase assembled pilus of the Gram-positive Enterococci, bind to fibrinogen on implanted catheters to promote biofilm formation and catheter-associated UTIs (CAUTIs). In collaboration with Project 1 this CORE will use state of the art high throughput cloning and selection methods to generate mAbs against all of these adhesins and evaluate their potential as antibiotic sparing therapeutics. These mAbs will also elucidate important epitopes for future anti-adhesin vaccine design. Further, all of the CUP adhesive pili are assembled by homologous systems involving a periplasmic chaperone and and outer membrane usher. The usher is a five domain assembly platform for CUP pilus polymerization and its transmembrane domain functions as a pore for extrusion of the assembled fiber. This pore is gated by a plug domain that only exits the pore when the usher is assembly a pilus fiber. Thus, usher proteins will be targeted for mAb generation and mAbs will be selected for both their ability to bind and inhibit usher mediated pilus assembly and/or for the ability to “open” the usher pore in the absence of a growing pilus fiber. Having an open pore without a pilus fiber will render the bacteria sensitive to antibiotics from our existing arsenal that are not generally able to cross the Gram-negative outer membrane and thus are not generally efficacious against Gram-negative pathogens. Targeting of adhesins and their assembly has the advantage that mutations in the binding pocket of targeted adhesins or in critical assembly sites in the usher to prevent binding of the therapeutic to its target would likely destroy adhesin function and thus render the pathogen non-virulent. The mAbs generated have the promise to be a viable alternative for the treatment of antibiotic-resistant infecitons. Therapeutic mAbs have not yet been fully harnessed for treating infectious diseases, perhaps due to the historic success of antibiotics. However, with antibiotic resistance on the rise, it is time to apply these potentially antibiotic-sparing tools to infectious disease.

项目摘要/摘要摘要： 抗生素耐药性日益威胁人类健康，导致感染难以治疗，其中一些感染 多重耐药病原体对所有或大多数抗生素具有抗药性，为了应对这一日益严重的危机，需要采取这种措施。 U19 正在采取综合方法来产生和开发针对关键毒力的疗法 具体来说，该核心将与项目 1 和 2 结合起来，寻求 开发针对不同细菌粘附素的单克隆抗体 (mAb) 疗法，广泛研究表明 事实证明，它对于病原体与宿主组织和表面结合并抵抗其清除的能力至关重要。 尿路致病性大肠杆菌 (UPEC) 使用尖端带有粘附素蛋白的 CUP 粘附菌毛来与多种人类结合 组织，包括未发炎的膀胱、发炎的膀胱、肾脏、导尿管和 此外，克雷伯菌表达一种甘露糖结合 FimH 粘附素，这对胃肠道 (GIT) 至关重要。 克雷伯菌引起尿路感染的能力此外，鲍曼不动杆菌中的 CUP 粘附素、CupD 和 PrpD 也是如此。 以及革兰氏阳性肠球菌分选酶组装菌毛顶端的 EbpA 粘附素，结合 植入导管上的纤维蛋白原可促进生物膜形成和导管相关性尿路感染 (CAUTI)。 与项目 1 合作，该核心将使用最先进的高通量克隆和选择方法 产生针对所有这些粘附素的单克隆抗体，并评估它们作为抗生素节约疗法的潜力。 这些单克隆抗体还将阐明未来抗粘附素疫苗设计的重要表位。 CUP 粘附菌毛由周质伴侣同源系统组装而成，并涉及和外部 膜引导器是用于 CUP 菌毛聚合及其聚合的五域组装平台。 跨膜结构域充当用于挤出组装纤维的孔，该孔由塞子控制。 仅当引导者组装菌毛纤维时才退出孔的结构域，因此引导蛋白将成为目标。 用于 mAb 生成，并且将根据 mAb 结合和抑制 usher 介导的菌毛的能力来选择 组装和/或在没有生长的菌毛纤维的情况下“打开”引导孔的能力。 没有菌毛纤维的毛孔将使细菌对我们现有的抗生素敏感，而这些抗生素对我们现有的抗生素不敏感。 通常能够穿过革兰氏阴性外膜，因此通常不能有效对抗 粘附素及其组装的靶向革兰氏阴性病原体具有突变的优点。 目标粘附素的结合口袋或引座中的关键组装位点，以防止结合 针对其靶标的治疗可能会破坏粘附素功能，从而使病原体失去毒性。 产生的单克隆抗体有望成为治疗抗生素耐药性感染的可行替代方案。 治疗性单克隆抗体尚未完全用于治疗传染病，这可能是由于 然而，随着抗生素耐药性的增加，是时候应用这些抗生素了。 传染病的潜在抗生素节约工具。