Engineering synthetic phages against pathogenic E. coli as an innovative tool for phage therapy

对患者安全的一个主要挑战是对抗生素具有耐药性的革兰氏阴性菌引起的医院感染。一种定义明确的这种细菌菌株是大肠杆菌（E. coli） O18:K1:H7，它是烧伤患者继发感染、新生儿脑膜炎和败血症以及急性膀胱炎的原因。解决这个问题的一个可能的办法是使用噬菌体作为抗菌剂。噬菌体是感染并杀死细菌的病毒。它们对目标细菌表现出高度特异性，同时对宿主的副作用最小，因此它们有可能用于治疗人类细菌感染。然而，对噬菌体治疗仍存在担忧，如免疫反应的可能性、噬菌体快速释放毒素以及临床情况下剂量确定的困难。此外，由于该领域的挑战，人们对噬菌体治疗背后的细胞生物学知之甚少，这也阻碍了噬菌体治疗的快速发展。该研究计划的主要目的是设计一个模型系统，作为噬菌体治疗的工具，该模型系统由三部分组成：能够靶向已知病原体的合成噬菌体，病原体（大肠杆菌O18:K1:H7），以及用于测试噬菌体-细菌相互作用的哺乳动物细胞，模拟人体条件。该系统经适当验证后，可进一步用于研究，为安全噬菌体疗法建立一个有希望的概念证明，该疗法可治疗由目标病原体引起的烧伤患者感染、新生儿脑膜炎或急性膀胱炎等疾病。结合分子生物学、合成生物学和显微学方法将使我能够实现这一目标。这项研究将在华威大学生命科学学院的Alfonso Jaramillo教授的实验室进行。根据2014年卓越研究框架，华威大学在英国排名第七。生命科学学院被评为世界领先（80%的产出被评为世界领先或国际优秀）。

SummaryA major challenge to patient safety is the hospital infections caused by Gram-negative bacteria that are resistant to antibiotics. A well-defined bacterial strain of this kind is Escherichia coli (E. coli) O18:K1:H7, which is responsible for secondary infections in burn patients, neonatal meningitis and sepsis, and acute cystitis. One of the possible solutions to this problem is the use of bacteriophages as antimicrobial agents. Bacteriophages are viruses that infect and kill bacteria. They show high specificity to their bacterial target, while having minimal side effects on the host, so they can potentially be used to treat bacterial infections in humans. However, there are still concerns for phage therapy, over the potential for immune response, rapid toxin release by the phages and difficulty of dose determination in clinical situations. Additionally, little is known about the cell biology underlying phage therapy, due to the challenges in the field, so that has been an obstacle in the rapid progress of phage therapy.The key aim of the research proposal is to engineer a model system as a tool for phage therapy consisting of 3 parts: a synthetic phage able to target a well-known pathogen, the pathogen (E.coli O18:K1:H7), and mammalian cells to test the phage-bacterium interplay mimicking the conditions of the human body. This system, with proper validation, can be used further for studies, establishing a promising proof of concept for safe phage therapy, which can treat conditions such as infection in burn wound patients, neonatal meningitis or acute cystitis, caused by the target pathogen. A combination of molecular biology, synthetic biology and microscopy methods will be enable me to achieve the objectives. The research will be undertaken in the School of Life Sciences, University of Warwick, in the lab of Professor Alfonso Jaramillo. The University of Warwick has been ranked as the 7th top institutions within the UK according to the Research Excellence Framework 2014. The School of Life Sciences was rated as world-leading (80% of its outputs were rated as world leading or internationally excellent).