Does CRISPR-Cas immunity limit the effectiveness of phage therapy?

CRISPR-Cas 免疫是否会限制噬菌体疗法的有效性？

• 批准号: BB/T014342/1
• 负责人: Michael Brockhurst
• 金额: $ 89.22万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FT014342%2F1
• 关键词: Does; CRISPR; Cas; immunity; limit

Globally rising rates of resistance against all major classes of chemotherapeutic antibiotics has renewed interest in the therapeutic use of phages to treat bacterial infections. As with antibiotic treatments, the emergence of bacterial resistance against phage treatments could also limit the long-term sustainability of phage-based therapeutics. We currently lack a mechanistic understanding of how bacteria evolve resistance against combinations of multiple phages, limiting our ability to design phage combinations that limit resistance. Importantly, bacteria possess multiple distinct mechanisms to defend against phage infection, including surface receptor modification (SRM) and CRISPR-Cas. These distinct mechanisms are favoured under different conditions which are likely to vary at the site of infection, but the importance of CRISPR-Cas for the evolution of resistance against phage therapy is unknown. We will use laboratory evolution experiments to determine how CRISPR-Cas immunity affects the evolution of multi-phage resistance across infection-relevant environmental conditions predicted to tip the balance from SRM to CRISPR-Cas immunity. We will then test if, by enhancing multi-phage resistance evolution, CRISPR-Cas immunity reduces the efficacy of phage therapy in vivo using a mouse model of acute respiratory infection. These experiments will develop and test an evolutionary framework for understanding how CRISPR-Cas affects multi-phage resistance evolution and the efficacy of phage therapy.

全球范围内对所有主要化疗抗生素类别的耐药率不断上升，重新引发了人们对噬菌体治疗细菌感染的兴趣。与抗生素治疗一样，细菌对噬菌体治疗产生耐药性的出现也可能限制噬菌体治疗的长期可持续性。目前，我们对细菌如何进化出对多种噬菌体组合的抗性缺乏机制了解，这限制了我们设计限制抗性的噬菌体组合的能力。重要的是，细菌拥有多种不同的机制来防御噬菌体感染，包括表面受体修饰 (SRM) 和 CRISPR-Cas。这些不同的机制在不同的条件下受到青睐，而感染部位可能会有所不同，但 CRISPR-Cas 对于噬菌体治疗耐药性进化的重要性尚不清楚。我们将使用实验室进化实验来确定 CRISPR-Cas 免疫如何影响多噬菌体抗性在感染相关环境条件下的进化，预计会扭转从 SRM 到 CRISPR-Cas 免疫的平衡。然后，我们将使用急性呼吸道感染小鼠模型来测试，通过增强多噬菌体抗性进化，CRISPR-Cas免疫是否会降低体内噬菌体疗法的功效。这些实验将开发和测试一个进化框架，以了解 CRISPR-Cas 如何影响多噬菌体抗性进化和噬菌体治疗的功效。

项目成果

Parallel evolution of Pseudomonas aeruginosa phage resistance and virulence loss in response to phage treatment in vivo and in vitro.

• DOI: 10.7554/elife.73679
• 发表时间: 2022-02-21
• 期刊: eLife
• 影响因子: 7.7
• 作者: Castledine M;Padfield D;Sierocinski P;Soria Pascual J;Hughes A;Mäkinen L;Friman VP;Pirnay JP;Merabishvili M;de Vos D;Buckling A
• 通讯作者: Buckling A