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

描述（由申请人提供）：我们的总体目标是开发我们的杀菌性R型绿脓菌素靶向大肠杆菌O157：H7，作为抗感染和污染的特异性治疗剂。肠出血性大肠大肠杆菌（EHEC）是一组食物和水传播的病原体，可引起从非血性腹泻到溶血性尿毒综合征（HUS）的疾病。e.大肠杆菌O157：H7可引起大规模暴发和散发感染，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，即R型绿脓菌素是高度特异性和高效的杀菌蛋白复合物，已被证明对动物中的全身性细菌感染有效。虽然对它们的靶生物具有高度特异性，但这些杀菌剂可以通过用噬菌体的相关尾蛋白修饰它们的尾纤维而重新靶向其他细菌。我们领先的候选产品AvR2-V10由噬菌体的尾部纤维组成。V10融合至R2型绿脓菌素，使得嵌合颗粒具有针对所有测试的O157 E的杀菌特异性。大肠杆菌菌株-即噬菌体的特异性。在本工作中，我们将进一步开发AvR 2-V10作为抗大肠杆菌的药物。coli O157：H7菌株。我们假设R型绿脓菌素可以在大肠杆菌中合成和产生。coliBL21系统，获得了有效的抗E.大肠杆菌O157：H7感染。为了实现我们的目标，我们将通过在E. coli BL21，以使其符合GLP标准（特定目标#1）。我们将在模拟与O157：H7感染相关的人类病理学的幼兔模型中评价其疗效（特定目标#2）。该候选产品预期用作特异性靶向大肠杆菌O157的治疗或预防药物，并可在流行病（意外或故意大规模感染）的情况下提供有效反应。公共卫生相关性：E。大肠杆菌O157可导致严重的肠道感染和潜在的死亡，并以生物恐怖袭击或意外食物或水传播爆发的形式对公共卫生构成威胁。我们利用平台技术设计了一种有效的杀菌剂AvR2-V10，专门针对大肠杆菌O157。目前正在开发这种药剂，作为治疗或预防大肠杆菌O157感染或污染的药物，在流行病的情况下提供有效的反应。在此，我们建议开发一种简单、高产的生产方法，并在家兔模型中测试其功效。

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.