TB Phage therapy: Optimizing delivery methods of mycobacteriophages to target intracellular Mycobacterium tuberculosis

摘要 结核病仍然是一个主要的全球健康问题，造成120万人死亡和10多万人死亡。 每年新增上百万个病例。20世纪80年代末开始的艾滋病毒流行导致免疫 在人体内抑制并明显增加抗药性的出现，延长 治疗持续时间。不祥的是，结核病的病原体--结核分枝杆菌的菌株 对目前可获得的极难治疗的结核病药物中的许多(如果不是全部)具有抗药性。作为一种 作为药物的替代品，分支杆菌噬菌体--杀死结核分枝杆菌的病毒--被认为是 可能用于治疗结核病的药物。然而，以前的研究在哺乳动物身上取得的成功有限。 主持人。这笔赠款的目的是阐明为什么噬菌体疗法不起作用。我们假设 主要障碍是分枝杆菌噬菌体无法接触到结核分枝杆菌细胞。 它们生活在细胞内。我们建议开发新的策略，将分枝杆菌噬菌体运送到 结核分枝杆菌所在的细胞内隔室。一种方法是设计 分枝杆菌噬菌体衣壳蛋白，使我们可以添加巨噬细胞递送配体来促进细胞 领悟。此外，我们计划探索使用污垢分枝杆菌作为特洛伊木马 细胞内结核分枝杆菌的递送载体。这些研究将通过我们的能力得到加强 产生表达荧光报告蛋白的分枝杆菌噬菌体。为了向公众传递 分枝杆菌噬菌体对肺部细胞内部位的定位，我们计划采用喷雾干燥 分枝杆菌噬菌体粉末。我们还计划测试分支杆菌噬菌体可能作为 在小鼠模型中使用现有结核病药物的辅助治疗。

Abstract Tuberculosis (TB) remains as a major global health problem causing 1.2 million deaths and over 10 million new cases each year. The onset of the HIV epidemic beginning in the late 1980s led to immune suppression in humans and significantly increased the emergence of drug resistance, lengthening treatment duration. Ominously, there are strains of M. tuberculosis, the causative agent of TB, that are resistant to many, if not all, of the currently available TB drugs that are extremely difficult to treat. As an alternative to drugs, mycobacteriophages - viruses that kill M. tuberculosis, have been considered as possible agents to treat tuberculosis. However, previous studies have had limited success in mammalian hosts. The goal of this grant is to elucidate why phage therapy fails to work. We hypothesized that the major obstacle is mycobacteriophages are unable to come into contact with the M. tuberculosis cells that live intracellularly. We propose to develop novel strategies to deliver mycobacteriophages to the intracellular compartments in which M. tuberculosis resides. One approach would be to engineer mycobacteriophage capsid proteins so that we can add macrophage delivery ligands to promote cellular uptake. In addition, we plan to explore the use of a Mycobacterium smegmatis strain as a Trojan horse delivery vector to the intracellular M. tuberculosis. These studies will be enhanced by our ability to generate mycobacteriophages that express fluorescent reporter proteins. In order to deliver the mycobacteriophages to the lungs, the site of intracellular localization, we plan to use spray dried mycobacteriophage powders. We also plan to test the hypothesis that mycobacteriophages may work as an adjunct therapy with existing TB drugs in mouse models.