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: 10162826
• 负责人: Ali Hassan Ellebedy
• 金额: $ 36.23万
• 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/10162826
• 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 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; Implant; Infection; Interdisciplinary Study; 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; Pilum; 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; host colonization; human tissue; innovation; multi-drug resistant pathogen; multidrug-resistant Pseudomonas aeruginosa; 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： 抗生素耐药性越来越对人类健康构成威胁，导致难以治疗感染，其中一些 多药的病原体对所有或大多数抗生素具有抗性。为了打击这一日益严峻的危机，这 U19正在采用综合方法来针对关键病毒的治疗剂的产生和开发 细菌的机理。具体而言，该核心将与项目1和2的集成方法中 开发针对不同细菌粘附素的单克隆抗体（MAB）疗法，广泛的研究具有 证明对病原体结合和抵抗从宿主组织和表面的去除至关重要。 肝病大肠杆菌（UPEC）使用带有粘合剂蛋白倾斜的杯子胶粘剂与多种人结合 包括非炎症的膀胱，发炎的膀胱，肾脏，尿导管和 胃肠道（GIT）。此外，克雷伯氏菌表达了一种甘露糖结合FIMH粘附素，这对于 克雷伯氏菌引起尿路感染的能力。此外，杯子粘附在鲍马尼（Baumanii），CUPD和PRPD中， 以及在革兰氏阳性肠球菌组装的吡咯的尖端上的EBPA粘合剂，与 植入导管上的纤维蛋白原以促进生物膜形成和与导管相关的UTI（CATIS）。 与项目1的合作该核心将使用最先进的高吞吐量克隆和选择方法 为了对所有这些粘附剂产生mAb，并评估它们作为抗生素备用疗法的潜力。 这些mAB还将阐明重要的表位，以实现未来的抗粘着素疫苗设计。此外，所有 杯子胶粘剂由涉及周质伴侣和外部的同源系统组装 膜Usher。 Usher是一个五个用于杯钢琴聚合及其杯子的域组装平台 跨膜结构域充当伸展组装纤维的孔。这个孔由插头盖住 只有在迎接一个枕头纤维时才能退出孔的域。那将是usher蛋白的目标 将选择MAB的生成和mAb，以既可以结合和抑制Usher中介飞行员的能力 在没有生长的活塞纤维的情况下，组装和/或能够“打开”带孔的能力。开放 没有枕头纤维的毛孔会使我们现有武器纳的抗生素敏感的细菌不是 通常能够越过革兰氏阴性外膜，因此通常不有效 革兰氏阴性病原体。靶向粘附剂及其组装的优势是 靶向粘附器的结合口袋或在usher中的关键装配位点，以防止结合 对其靶标的治疗可能会破坏粘附功能，从而导致病原体非病毒病毒。 产生的mAB有望成为治疗抗生素耐药的infecitons的可行替代方法。 治疗性mABS尚未被完全利用用于治疗传染病，也许是由于 抗生素的历史成功。但是，随着抗生素抗性的上升，是时候应用这些 潜在的抗生素具有传染病的工具。