Aptamer-Enabled Modification of Bacteriophage Host Range

噬菌体宿主范围的适体修饰

• 批准号: 9802755
• 负责人: Hua Shi
• 金额: $ 23.18万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9802755
• 关键词: Affinity; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Bacteria; Bacterial Antibiotic Resistance; Bacterial Infections; Bacteriophage T4; Bacteriophages; Binding; Binding Proteins; Biological Assay; Biological Models; Bypass; Capsid; Cell surface; Cells; Cessation of life; Characteristics; Chemicals; Clinical Research; Collection; Conscious; Coupling; Cryoelectron Microscopy; DNA; DNA Shuffling; Development; Electron Microscopy; Escherichia coli; Face; Failure; Fiber; Genome; Genomic DNA; Goals; Imagery; Incentives; Individual; Infection; Intellectual Property; Knock-out; Lipopolysaccharides; Maintenance; Methods; Microscopy; Modification; Molecular Conformation; Morphology; Motion; Nature; OmpC protein; Periodicity; Phage Receptors; Pharmacologic Substance; Pharmacology; Plague; Preparation; Process; Property; Proteins; Protocols documentation; Public Health; RNA; Research; Resistance; Resources; Ribonucleases; Safety; Scheme; Secure; Series; Signal Transduction; Specificity; Superbug; Surface; System; Tail; Testing; Therapeutic; Thyroxine Receptors; Time; Uncertainty; United States National Institutes of Health; Untranslated RNA; Update; Virion; Virus; X-Ray Crystallography; aptamer; bactericide; base; combat; cost; design; drug discovery; experience; experimental study; improved; innovation; macromolecule; microbiota; mimicry; molecular scale; nanoscale; novel therapeutics; operation; particle; pathogenic bacteria; peer; preclinical study; prophylactic; receptor; receptor binding; research and development; response; targeted treatment; weapons

Project Summary The term “superbug” conjures up in public consciousness the imagery of nasty bacteria recalcitrant to antibiotic treatments, a public health threat further heightened by the dwindling supply of new antibiotics in the development pipeline. Imprudent use of broad-spectrum antibiotics is also harmful to the beneficial microbiota. With the renewed urgency, the US National Institutes of Health listed phage therapy as one of seven prongs in its plan to combat antibiotic resistance. Bacteriophages (phages) are viruses that only infect and kill bacteria. Phage therapy is the targeted application of phages as bactericidal agent to treat bacterial infections. However, to better respond to emerging and evolving bacterial pathogens, all therapeutic and prophylactic phage products will require periodic updates, which will inevitably incur further cost in product maintenance. The unpatentability of “product of nature” further reduces incentives for commercial entities to participate in the research and development (R&D) to realize the full potential of phage therapy. Here we propose an innovative approach to bypass the required updates that can plague the conventional whole-phage products, and at the same time provide an incentive for R&D with an intellectual property that is easily securable and defendable. Our proposal is made possible by recent studies, revealing intricate steps involved in host recognition by a phage and how the signal of recognition is able to trigger the preprogrammed conformational changes in the phage tail that eventually leads to the delivery of phage DNA genome into the infected bacterial cell, thus killing it. We take advantages of the extensively studied classics, T4 and T7, and their host, Escherichia coli, as the model system to test the hypothesis that an artificially tethered phage can be triggered to deliver its DNA genome into a non-host cell. In this scheme, phage tethering is achieved by chemically stable RNA aptamers that specifically bind to the phage tail fiber, the receptor-binding protein of the phage, and those that bind to the surface macromolecules of E. coli. By coupling together these two types of aptamers, we can bring a phage to close physical proximity of the cell surface, a pre-requisite for a successful infection. Based on the hypothesis, the Brownian motion, experienced by an attached phage, will trigger the preprogrammed deployment of the proteins involved in phage tail such that the DNA genome can be delivered. We will use the well-established molecular breeding process, called SELEX, to generate the required aptamers. Our proposed study represents a unique solution to the problems—especially the scalability and the patentability—encountered by using phage as a therapeutic and prophylactic agent against bacterial infection. If the result is encouraging from this proof-of-concept study, we believe we will be able to open an entirely new and impactful revenue of research.

项目摘要 一词“超级细菌”一词在公共意识中引起了讨厌的细菌的形象， 抗生素治疗是一种公共卫生威胁，这是由于新的抗生素的供应不断增长 发展管道。广谱抗生素的不当使用也对有益的微生物群也有害。 随着新的紧迫性，美国国立卫生研究院将噬菌体疗法列为 它打击抗生素耐药性的计划。噬菌体（噬菌体）是仅感染并杀死细菌的病毒。 噬菌体治疗是噬菌体作为细菌感染的细菌剂的靶向应用。然而， 为了更好地应对新兴和进化的细菌病原体，所有治疗和预防性噬菌体 产品将需要定期更新，这将不可避免地产生产品维护的进一步成本。 “自然产品”的不遵守性进一步减少了商业实体参与的激励措施 研发（R＆D），以实现噬菌体疗法的全部潜力。 在这里，我们提出了一种创新的方法，以绕过可能困扰的所需更新 传统的全境产品，同时为R＆D提供了一种诱因 易于确保和防御的财产。最近的研究使我们的建议成为可能 通过噬菌体涉及主机识别的复杂步骤，以及识别信号如何触发 噬菌体尾巴的预编程会议变化有时会导致噬菌体DNA的递送 基因组进入被感染的细菌细胞，从而杀死它。我们利用广泛研究的经典， T4和T7及其宿主Escherichia Coli，作为测试人为假设的模型系统 可以触发束缚噬菌体将其DNA基因组传递到非宿主细胞中。 在该方案中，噬菌体束缚是通过专门结合的化学稳定RNA适体实现的 噬菌体尾纤维，噬菌体的接收器结合蛋白以及与表面结合的蛋白 大肠杆菌的大分子。通过将这两种类型的适体耦合在一起，我们可以使噬菌体关闭 细胞表面的物理接近，这是成功感染的先决条件。基于假设 布朗尼运动是由附带的噬菌体经历的，将触发预编程的部署 参与噬菌体尾巴的蛋白质可以传递DNA基因组。我们将使用良好的 分子繁殖过程称为SELEX，以生成所需的适体。 我们提出的研究代表了解决问题的独特解决方案，尤其是可伸缩性和 可专利性 - 通过将噬菌体用作针对细菌感染的治疗和预防剂。 如果结果在这项概念证明的研究中令人鼓舞，我们相信我们将能够打开一个全新的 和有影响力的研究收入。