Burke Kevin A, Peters Tracey L, Kirillina Olga A, Urick Caitlin D, Walton Bertran D, Bird Jordan T, Mzhavia Nino, Georges Martin O, Lertsethtakarn Paphavee, Musila Lillian A, Nikolich Mikeljon P, Filippov Andrey A
Wound Infections Department, Bacterial Diseases Branch, Center for Infectious Diseases Research, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA.
Department of Entomology, College of Agricultural, Human and Natural Resource Sciences, Washington State University, Pullman, WA 99164, USA.
Int J Mol Sci. 2025 Aug 6;26(15):7597. doi: 10.3390/ijms26157597.
Multidrug-resistant (MDR) strains of present an acute threat as they continue to disseminate globally. Phage therapy has shown promise as a powerful approach to combat MDR infections, but narrow phage host ranges make development of broad acting therapeutics more challenging. The goal of this effort was to use in vitro directed evolution (the "Appelmans protocol") to isolate phages with broader host ranges for improved therapeutic cocktails. Five myophages in the genus (family ) with complementary activity were mixed and passaged against a panel of 11 bacterial strains including a permissive host and phage-resistant clinical isolates. Following multiple rounds of training, we collected phage variants displaying altered specificity or expanded host ranges compared with parental phages when tested against a 100 strain diversity panel of . Some phage variants gained the ability to lyse previously phage-resistant strains but lost activity towards previously phage-susceptible strains, while several variants had expanded activity. Whole-genome sequencing identified mutations and recombination events impacting genes associated with host tropism including tail fiber genes that most likely underlie the observed changes in host ranges. Evolved phages with broader activity are promising candidates for improved therapeutic phage cocktails.
多重耐药(MDR)菌株在全球范围内持续传播,构成了严重威胁。噬菌体疗法已显示出作为对抗MDR感染的有力方法的潜力,但噬菌体宿主范围狭窄使得开发广谱治疗药物更具挑战性。这项工作的目标是利用体外定向进化(“阿佩尔曼斯方案”)来分离具有更广泛宿主范围的噬菌体,以改进治疗性鸡尾酒制剂。将5种具有互补活性的肌尾噬菌体属(肌尾噬菌体科)噬菌体混合,并使其针对一组11种细菌菌株传代,这些菌株包括一个敏感宿主和噬菌体抗性临床分离株。经过多轮培养后,当针对100株多样性的菌株进行测试时,我们收集到了与亲本噬菌体相比显示出特异性改变或宿主范围扩大的噬菌体变体。一些噬菌体变体获得了裂解先前抗噬菌体菌株的能力,但失去了对先前敏感菌株的活性,而有几个变体的活性有所扩大。全基因组测序确定了影响与宿主嗜性相关基因的突变和重组事件,包括尾丝基因,这些基因很可能是观察到的宿主范围变化的基础。具有更广泛活性的进化噬菌体有望成为改进治疗性噬菌体鸡尾酒制剂的候选者。
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