Biological validation of phage host-range identified by proximity guided metagenomics

摘要 噬菌体是一种感染细菌的病毒，有可能被用作治疗传染性疾病的临床工具。 疾病和肠道微生物组相关疾病。了解噬菌体与宿主的相互作用可以提高 使用噬菌体作为一种治疗选择，并可能导致新疗法的开发。然而， 目前研究这些相互作用的方法是低通量和基于文化的，限制了在 噬菌体疗法研究。邻近连接测序，它使用体内噬菌体和 宿主基因组，是一种高通量测序工具，可用于读出这些关联。 然而，还没有进行严格的实验验证来验证这些相互作用，也没有 描述该方法的分析性能。 该项目的目标是建立邻近连接测序作为研究的可靠科学工具 噬菌体-寄主联合。我们建议使用金标准微生物方法和 高级测序工具，可严格验证和定义邻近连接测序的性能 用于研究噬菌体与宿主的相互作用。我们将确定邻近结扎术的敏感性和特异性。 使用定义的微生物群落和自然粪便微生物群落进行测序。此外, 我们将评估邻近连接识别的噬菌体-宿主联合在复合体中的阳性预测价值 使用噬菌体分离技术的微生物群落。这些实验将提供关键的统计数据 可靠性参数，解锁了一种高通量、不依赖培养的噬菌体宿主鉴定方法 互动。这种方法将在基于噬菌体的应用程序中启用新途径，包括 基于噬菌体治疗感染性疾病、肠道生物失调和靶向操作的研究进展 微生物群。

Abstract Phages are viruses that infect bacteria and have the potential to be used as a clinical tool for treating infectious diseases and gut microbiome-related disorders. Understanding phage-host interactions can improve the ability to use phages as a treatment option, and potentially lead to the development of new therapies. However, current methods for studying these interactions are low-throughput and culture-based, limiting progress in phage therapy research. Proximity ligation sequencing, which uses in vivo crosslinking between phage and host genomes, is a high-throughput sequencing tool which can be used to read out these associations. However, no rigorous experimental validation has yet been performed to verify these interactions, nor to characterize the analytical performance of this approach. The objective of this project is to establish proximity ligation sequencing as a reliable scientific tool for studying phage-host associations. We propose to use a combination of gold-standard microbiological methods and advanced sequencing tools to rigorously validate and define the performance of proximity ligation sequencing for studying phage-host interactions. We will determine the sensitivity and specificity of proximity ligation sequencing using both defined microbial communities and natural fecal microbiome communities. In addition, we will estimate the positive predictive value of proximity ligation-identified phage-host associations in complex microbial communities using phage isolation techniques. These experiments will provide key statistical parameters of reliability, unlocking a high-throughput, culture-independent method of characterizing phage-host interactions. This approach would enable new avenues in phage-based applications, including the development of phage-based therapies for infectious disease, gut dysbiosis, and the targeted manipulation of microbiomes.