Development of Novel Bacteriophages Targeting Enteric Bacterial Pathogens

摘要 这份R21提案的重点是开发针对肠道细菌病原体的新疗法， 尤其是志贺氏菌。这些病原体导致数百万人感染和惊人的死亡人数。 尽管在清洁水、卫生设施和口服补液疗法等方面取得了进展，但全球每年都有新的人口流动。 感染是由于细菌侵袭胃肠道衬里的上皮细胞造成的，而且主要是 发生在发展中国家的五岁以下儿童。在工业化国家，感染是 与日托中心、食源性和水源性疫情有关。尽管进行了几十年的研究，但没有 针对志贺氏菌的有效疫苗。此外，抗生素耐药性的惊人增长使情况变得复杂起来。 治疗。这项建议寻求建立在最新发现的基础上，以开发和评估病原体特异性 噬菌体，感染细菌的病毒，用于未来的治疗开发。这个项目的长期目标是 是利用生物工程和合成生物学来开发可用于治疗的新型噬菌体 治疗感染性腹泻而不损害健康的共生细菌种群的选择 胃肠道。这项建议的具体目标是：第一，利用生物工程来提高我们的 原型噬菌体，并随后测试用于病原体特异性裂解的增强候选；以及第二， 进一步改造原型噬菌体以扩大寄主范围并延迟抗噬菌体的出现 细菌来规避天然噬菌体的常见问题。拟议的研究包括使用 模拟人类特有环境的创新方法、技术和感染模型 胃肠道。该提案得到了一个研究团队的进一步加强，该团队结合了 噬菌体生物学、生物工程、细菌发病机制、粘膜生物学和有机物衍生 感染模型。从AIMS获得的数据将导致新疗法的开发，这可能证明 在抗菌素耐药性猖獗和缺乏有效疫苗的时代，要在临床上有效。 1

Abstract This R21 proposal focuses on the development of novel therapeutics against enteric bacterial pathogens, specifically the Shigella species. The pathogens cause millions of infections and a staggering number of deaths around the globe each year despite advances such as clean water, sanitation, and oral rehydration therapy. Infection is due to bacterial invasion of the epithelial cells lining the gastrointestinal tract, and predominately occurs in children under the age of five years in developing countries. In industrialized nations, infections are linked to daycare center, foodborne, and waterborne outbreaks. Despite decades of research, there are no effective vaccines against Shigella. Furthermore, alarming increases in antibiotic resistance have complicated treatment. This proposal seeks to build upon recent findings to develop and evaluate pathogen-specific bacteriophages, viruses that infect bacteria, for future therapeutic development. The long-term goal of this project is to utilize bioengineering and synthetic biology to develop novel bacteriophages that can be used as therapeutic options to treat infectious diarrhea without harming the healthy commensal bacterial population of the gastrointestinal tract. The specific aims of this proposal are to first, utilize bioengineering to enhance our prototype bacteriophage and subsequently test the enhanced candidate for pathogen-specific lysis; and second, further engineer the prototype phage to expand the host range and delay the emergence of phage-resistant bacteria to circumvent common issues with natural bacteriophages. The proposed research includes the use of innovative approaches, technologies, and infection models that mimic the human-specific environment of the gastrointestinal tract. The proposal is further enhanced by a research team that combines the expertise of bacteriophage biology, bioengineering, bacterial pathogenesis, and mucosal biology, and organoid-derived infection models. Data obtained from the aims will lead to the development of novel therapeutics that could prove to be clinically effective in an age of rampant antimicrobial resistance and a lack of effective vaccines. 1