Engineered R-type pyocins as bactericidal agents against E.coli O157:H7

工程化 R 型脓毒素作为针对大肠杆菌 O157:H7 的杀菌剂

• 批准号: 8312148
• 负责人: DEAN M SCHOLL
• 金额: $ 30.85万
• 依托单位: AVIDBIOTICS CORPORATION
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-02 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8312148
• 关键词: Accounting; Acute Kidney Failure; Advanced Development; Animal Model; Animals; Antibiotics; Bacteria; Bacterial Infections; Bacteriophages; Binding; Cell Surface Receptors; Cessation of life; Child; Clinical Trials; Development; Diarrhea; Disease Outbreaks; Dose; Engineering; Epidemic; Escherichia coli; Escherichia coli EHEC; Escherichia coli Infections; Escherichia coli O157; Feces; Fiber; Food; Future; Genome; Goals; Health; Hemolytic-Uremic Syndrome; Human; Human Pathology; Hydration status; Individual; Infant; Infection; Intervention; Intestines; Methods; Mitomycins; Modeling; Molecular Weight; Mus; Mutagens; Organism; Oryctolagus cuniculus; Persons; Preclinical Testing; Procedures; Process; Production; Protein Subunits; Proteins; Protocols documentation; Pseudomonas aeruginosa; Public Health; Quality Control; Risk; Salmonella; Specificity; System; Tail; Technology; Testing; Therapeutic; Treatment Efficacy; United States; Work; Yersinia pestis; bactericide; bacteriocin; base; efficacy testing; foodborne; in vivo; killings; particle; pathogen; pathogenic bacteria; prophylactic; protein complex; public health relevance; receptor binding; response; scale up; waterborne

DESCRIPTION (provided by applicant): Our overall Goal is to develop our bactericidal R-type pyocin targeting E.coli O157:H7, as a specific therapeutic against infection and contamination. Enterohemorrhagic E. coli (EHEC) are a group of food- and waterborne pathogens that cause illnesses ranging from non-bloody diarrhea to hemolytic uremic syndrome (HUS). E. coli O157:H7 causes both large outbreaks as well as sporadic infections, and thus poses a threat to public health. The work proposed here is based on our findings that R-type pyocins are highly specific and highly potent bactericidal protein complexes that have been shown to be efficacious for systemic bacterial infections in animals. While highly specific to their target organism, these bactericidal agents can be retargeted to other bacteria by modifying their tail fibers with the related tail proteins of bacteriophages. Our leading product candidate, AvR2-V10, consists of the tail fiber of the bacteriophage ?V10 fused to an R2-type pyocin, such that the chimeric particle has the bactericidal specificity against all tested O157 E. coli strains - i.e. the specificity of the bacteriophage. In the work proposed here, we will advance the development of AvR2-V10 as an anti E. coli O157:H7 agent. We hypothesize that R-type pyocins can be synthesized and produced in an E. coli BL21 system, yielding an effective therapeutic against E. coli O157:H7 infection. In order to achieve our goals, we will conduct preclinical testing of AvR2-V10 by producing it in a regulatable manner in E. coli BL21, such that it abides to GLP standards (Specific Aim #1). We will evaluate its efficacy in an infant rabbit model which mimics the human pathology associated with O157:H7 infection (Specific Aim #2). The product candidate is intended to be used as a therapeutic or prophylactic that specifically targets E.coli O157 and can offer a potent response in cases of epidemic, either accidental or intentional mass infection. PUBLIC HEALTH RELEVANCE: E. coli O157 can cause severe intestinal infections and potentially death, and poses a threat to public health in the form of a bioterrorist attack or accidental food or waterborne outbreak. We have used our platform technology to engineer a potent bactericidal agent, AvR2-V10, to specifically target E.coli O157. This agent is now being developed as a therapeutic or prophylactic against E.coli O157 infection or contamination, offering a potent response in cases of epidemic. Herein, we propose to develop a simple, high yield production method for this agent, and test it's efficacy in a rabbit model.

描述（由申请人提供）：我们的总体目标是开发针对大肠杆菌O157:H7的杀菌剂r型脓毒素，作为对抗感染和污染的特异性治疗药物。肠出血性大肠杆菌（EHEC）是一组通过食物和水传播的病原体，可引起从非出血性腹泻到溶血性尿毒症综合征（HUS）等各种疾病。大肠杆菌O157:H7既可引起大规模暴发，也可引起散发感染，从而对公共卫生构成威胁。本文提出的工作是基于我们的发现，r型脓毒蛋白是高度特异性和高效的杀菌蛋白复合物，已被证明对动物全身细菌感染有效。虽然这些杀菌剂对目标生物具有高度特异性，但它们可以通过用噬菌体的相关尾部蛋白修饰其尾部纤维来重新靶向其他细菌。我们的主要候选产品AvR2-V10由噬菌体的尾纤维组成。V10与r2型脓毒素融合，使得嵌合颗粒对所有测试的O157大肠杆菌菌株具有杀菌特异性，即噬菌体的特异性。在本文的工作中，我们将推进AvR2-V10作为抗大肠杆菌O157:H7的药物的开发。我们假设r型脓毒杆菌素可以在大肠杆菌BL21系统中合成和生产，从而有效治疗大肠杆菌O157:H7感染。为了实现我们的目标，我们将通过在大肠杆菌BL21中以可调节的方式生产AvR2-V10来进行临床前测试，使其符合GLP标准（Specific Aim #1）。我们将在模拟人类与O157:H7感染相关病理的幼兔模型中评估其疗效（特异性目标#2）。该候选产品旨在作为一种治疗或预防药物，专门针对大肠杆菌O157，并可在意外或故意集体感染的流行情况下提供有效的反应。公共卫生相关性：大肠杆菌O157可引起严重的肠道感染并可能导致死亡，并以生物恐怖袭击或意外的食物或水传播疫情的形式对公共卫生构成威胁。我们利用我们的平台技术设计了一种强效杀菌剂AvR2-V10，专门针对大肠杆菌O157。目前正在开发这种药物，作为治疗或预防大肠杆菌O157感染或污染的药物，在发生流行病时提供强有力的应对措施。在此，我们建议开发一种简单、高产的生产方法，并在家兔模型上测试其功效。