铜绿假单胞菌的合成噬菌体的理性设计与重建

The prevalence of multidrug-resistant (MDR) pathogens has grown far worse in recent years with inappropriate use of antimicrobial drugs. There is an urgent need for the emergence of an alternative strategy for fighting MDR bacteria. The strong lytic activity of natural bacteriophage has promoted phage therapy as a potential alternative application in medicine. However, there are great challenges for the proven safety of therapeutic phages. In this project, we attempt to explore a new strategy of rational design and engineering the virulent bacteriophage of pseudomonas aeruginosa. We will firstly simplify the genome to obtain an artificial infectious phage based on the combination of rational design and semi-rational design and screening, and then introduce a genetic module into the trimmed genome to inducing the programmed cell death for control the growth of drug-resistant bacteria, which will replace the natural lysis components and avoid immunogenicity caused by cell lysis. From these studies, this proposal will establish a method of preparing synthetic phages with the controllable risk. The outcomes of this project will help to better understand the interactions between genetic structures and functions of bacteriophages, and provide new ideas for the phage therapy and the treatment of bacterial resistance.

细菌耐药性的问题愈发严重，寻找可替代抗生素的新型的治疗方法一直是应对耐药菌的主要研究策略。烈性噬菌体对细菌的天然裂解能力使其成为有望解决细菌耐药性问题的重要方案之一。然而天然噬菌体的安全性难以有效控制，使噬菌体治疗在临床应用上存在重大挑战。本项目拟以铜绿假单胞菌的天然烈性噬菌体为研究对象，发展一种基于理性设计的合成噬菌体的重建策略：1）通过理性设计和半理性设计与筛选实现基因组的精简，并重构具有感染能力的合成噬菌体；2）为避免细胞裂解带来的免疫风险，通过引入控制耐药菌生长的表达模块以替换天然裂解组分，实现细菌的程序性死亡，从而建立一种安全风险可控的合成噬菌体的制备策略。本项目的实施将有助于更好的理解噬菌体的基因组结构与功能关系，为噬菌体治疗和解决细菌耐药问题提供新思路。

细菌耐药性的问题愈发严重，寻找可替代抗生素的新型的治疗方法一直是应对耐药菌的主要研究策略。噬菌体对细菌的天然裂解能力使其成为有望解决细菌耐药性问题的重要方案之一。然而天然噬菌体的安全性难以有效控制，使噬菌体治疗在临床应用上存在重大挑战。本项目从筛选铜绿假单胞菌的耐药菌的特异性噬菌体出发，筛选了4株铜绿假单胞菌特异性噬菌体，探索和建立了两种噬菌体的改造策略：1）利用CRISPR-Cas基因编辑技术改造噬菌体；2）利用基因组合成技术合成噬菌体。本项目通过上述两种策略成功获得了铜绿假单胞菌的活性重组噬菌体，为铜绿假单胞菌的噬菌体治疗提供技术思路和实验基础。