Intrabodies that prevent or disrupt biofilms

防止或破坏生物膜的体内抗体

• 批准号: 8872844
• 负责人: BARBARA I KAZMIERCZAK
• 金额: $ 24.97万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8872844
• 关键词: Address; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antibodies; Antigens; Bacteria; Behavior; Binding; Biological Factors; Biology; Catheters; Chronic; Clinical; Collaborations; Cystic Fibrosis; Cytoplasm; Data; Development; Devices; Gene Targeting; Genes; Genetic Screening; Growth; Harvest; Heart Valves; Hereditary Disease; Human; Immune response; Immune system; Individual; Infection; Joint Prosthesis; Laboratories; Libraries; Lung; Maintenance; Medical Device; Microbial Biofilms; Modeling; Molecular Target; Mutation; Osteomyelitis; Pathogenesis; Patients; Phenotype; Pneumonia; Prevention; Proteins; Pseudomonas aeruginosa; Reagent; Recurrence; Role; Route; Skin; Staging; Stents; Therapeutic; Tracheostomy procedure; Tube; Urinary tract infection; base; bone prosthesis; clinical application; cystic fibrosis patients; endotracheal; genetic approach; mutant; novel; pathogen; periplasm; prevent; public health relevance; screening; small molecule; small molecule libraries; therapeutic target; tool; urinary; ventricular assist device

 DESCRIPTION (provided by applicant): The ability of Pseudomonas aeruginosa to grow as a biofilm on both biotic and abiotic materials underlies its ability to establish chronic infections nd to persist despite antibiotic treatment and a host immune response. Therefore a better understanding of how biofilms form, grow and disperse addresses not only fundamental aspects of P. aeruginosa pathogenesis, but also identifies potential bacterial targets for anti-biofilm therapies. This application proposes the novel use of intrabodies in phenotypic screens to identify molecules that will inhibit biofilm formation, promote biofilm dispersal or disrupt biofil integrity. Our library of intrabodies-small, unmodified single-chain camelid antibodies raised against a highly diverse pool of P. aeruginosa antigens- can be inducibly expressed inside bacteria and targeted to the cytoplasm or periplasm. The ability to induce intrabody expression allows phenotypic screens for both biofilm dispersal as well as formation to be carried out, an advantage over genetic approaches. The active intrabody provides a tool for direct identification of the disrupted "target"-a significant advantage over small molecule screening approaches-as it can be used to bind and purify its target, and even identify the part of the target relevant o its function. Therefore, we hypothesize that the application of intrabody phenotypic screening to P. aeruginosa biofilm biology will identify novel "targets" involved in biofilm formation and maintenance inaccessible to genetic screens, and also generate intrabody reagents uniquely suited to studying the mechanism of target action and its potential for development as a therapeutic target.

 描述（由申请方提供）：铜绿假单胞菌在生物和非生物材料上作为生物膜生长的能力是其建立慢性感染并在抗生素治疗和宿主免疫应答下持续存在的能力的基础。因此，更好地理解生物膜如何形成、生长和分散不仅解决了铜绿假单胞菌发病机制的基本方面，而且还确定了抗生物膜疗法的潜在细菌靶标。本申请提出了胞内抗体在表型筛选中的新用途，以鉴定将抑制生物膜形成、促进生物膜分散或破坏生物膜完整性的分子。我们的胞内抗体库-针对高度多样化的铜绿假单胞菌抗原库产生的小的未修饰的单链骆驼抗体-可以在细菌内诱导表达并靶向细胞质或周质。诱导胞内抗体表达的能力允许进行生物膜分散和形成的表型筛选，这是优于遗传方法的优势。活性胞内抗体提供了一种直接鉴定破坏的“靶标”的工具，这是一个优于小分子筛选方法的显著优势，因为它可以用于结合和纯化其靶标，甚至鉴定与其功能相关的靶标部分。因此，我们假设胞内抗体表型筛选对铜绿假单胞菌生物膜生物学的应用将鉴定涉及生物膜形成和维持的遗传筛选不可及的新“靶标”，并且还产生独特地适合于研究靶标作用机制及其作为治疗靶标开发的潜力的胞内抗体试剂。