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，导致难以治疗的感染，一些 对所有或大部分抗生素都有抗药性的多重耐药病原体。为了应对这场日益严重的危机，这 U19正在采取一种综合的方法来产生和开发针对关键毒力的疗法 细菌的机制。具体地说，这一核心将与项目1和2采取综合办法，力求 开发针对不同细菌粘附素的单抗(MAb)疗法，已有广泛研究 被证明对病原体结合到宿主组织和表面并抵抗从宿主组织和表面移除的能力至关重要。 尿路致病性大肠杆菌(UPEC)使用末端带有粘附素蛋白的杯状粘附性菌毛与多种人 组织包括未发炎的膀胱、发炎的膀胱、肾脏、导尿管和 胃肠道(GIT)。此外，克雷伯氏菌还表达一种甘露糖结合的FimH粘附素，这对 克雷伯氏菌引起尿路感染的能力。此外，鲍曼不动杆菌、CupD和PRPD中的CUP粘附素，AS 以及位于革兰氏阳性肠球菌分解酶组装菌毛末端的EBPA粘附素，结合到 植入导管上的纤维蛋白原可促进生物膜的形成和导管相关性尿路感染(CAUTI)。在……里面 与Project 1合作此核心将使用最先进的高通量克隆和选择方法 制备针对所有这些粘附素的单抗，并评估其作为抗生素节省疗法的潜力。 这些单抗还将阐明未来抗粘附素疫苗设计的重要表位。此外，所有的 杯状粘附性菌毛由含有周质伴侣和外膜的同源系统组装而成 膜引导器。引座器是杯状聚合的五个域组装平台，其 跨膜结构域起到挤出组装纤维的孔的作用。这个小孔是用塞子堵住的 只有在引座员组装毛发纤维时才会离开毛孔的区域。因此，USER蛋白将成为目标 对于mAb的产生和mAbb的选择是因为其结合和抑制引导者介导的菌毛的能力 组件和/或能够在没有生长的绒毛纤维的情况下打开引导器毛孔。敞开心扉 没有菌毛纤维的毛孔会使细菌对我们现有军火库中没有的抗生素敏感 通常能够穿过革兰氏阴性外膜，因此通常对 革兰氏阴性病原体。靶向粘附素及其组装的优势是 靶向粘附素的结合口袋或在引座器中的关键组装部位，以防止结合 对其靶标进行治疗可能会破坏粘附素功能，从而使病原体无毒。这个 产生的单抗有望成为治疗抗生素耐药性感染的一种可行的替代方案。 治疗性单抗尚未完全用于治疗传染病，可能是由于 抗生素取得了历史性的成功。然而，随着抗生素耐药性的上升，是时候应用这些 潜在的节省抗生素的工具，以预防传染病。