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.

全球对所有主要类别化学治疗抗生素的抗药性率上升的速率已开始对治疗细菌感染的治疗用途的兴趣。与抗生素治疗一样，对噬菌体治疗的细菌耐药性的出现也可能限制基于噬菌体的疗法的长期可持续性。目前，我们缺乏对细菌如何发​​展抗多种噬菌体组合的抗性的机械理解，从而限制了我们设计限制抗抗性的噬菌体组合的能力。重要的是，细菌具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条件下可能会在感染部位有所不同，但是CRISPR-CAS对抗噬菌体疗法的抵抗力的重要性尚不清楚。我们将使用实验室进化实验来确定CRISPR-CAS免疫如何影响相关的与感染相关的环境条件的多层耐药性演变，这些环境条件被预测，可以使从SRM到CRISPR-CAS免疫的平衡。然后，我们将通过使用急性呼吸道感染的小鼠模型来测试CRISPR-CAS免疫是否通过增强多型耐药性进化来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如何影响多型耐药性进化和噬菌体疗法的功效。

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