中文摘要：传统抗生素日益失效，耐多药（MDR）细菌出现，促使噬菌体疗法复兴。然而，细菌也会进化出噬菌体耐药性。噬菌体及其细菌宿主在自然界中共存，使二者为生存竞争而不断发生共同进化。本研究分离到几株铜绿假单胞菌临床菌株和感染它们的噬菌体。其中，肌病毒科家族的PIAS噬菌体（诱导抗生素敏感的噬菌体）可诱导铜绿假单胞菌耐药临床菌株的多步基因组缺失，使细菌宿主的药物外排系统受损。研究发现了两种类型的突变株系：具有SNP（单核苷酸多态性）和较小缺失的绿色突变株，以及具有较大（约250 kbp）基因组缺失的棕色突变株；证明了PIAS使用MexXY OprM系统引发感染；铜绿假单胞菌通过修饰或删除这些受体来阻止PIAS噬菌体感染。通过SNPs获得噬菌体耐药性的绿色突变体可以通过其尾部纤维蛋白突变的进化PIASs (E-PIASs)来克服。这些突变体通过完全丧失或大量修改噬菌体受体MexXY-OprM，否定其在抗生素耐药性中的重要作用而获得噬菌体耐药性。体外和体内研究表明，联合使用PIAS和抗生素可有效抑制铜绿假单胞菌的生长。噬菌体既可以消灭细菌，也可以诱导耐多药临床菌株对抗生素的敏感性。研究探讨了联合疗法作为治疗耐多药铜绿假单胞菌感染的替代方法的潜在用途。
外文摘要：The increasing ineffectiveness of traditional antibiotics and the rise of multidrug resistant (MDR) bacteria have necessitated the revival of bacteriophage (phage) therapy. However, bacteria might also evolve resistance against phages. Phages and their bacterial hosts coexist in nature, resulting in a continuous coevolutionary competition for survival. We have isolated several clinical strains of Pseudomonas aeruginosa and phages that infect them. Among these, the PIAS (Phage Induced Antibiotic Sensitivity) phage belonging to the Myoviridae family can induce multistep genomic deletion in drug-resistant clinical strains of P. aeruginosa, producing a compromised drug efflux system in the bacterial host. We identified two types of mutant lines in the process: green mutants with SNPs (single nucleotide polymorphisms) and smaller deletions and brown mutants with large (similar to 250 kbp) genomic deletion. We demonstrated that PIAS used the MexXY-OprM system to initiate the infection. P. aeruginosa clogged PIAS phage infection by either modifying or deleting these receptors. The green mutant gaining phage resistance by SNPs could be overcome by evolved PIASs (E-PIASs) with a mutation in its tail-fiber protein. Characterization of the mutant phages will provide a deeper understanding of phage-host interaction. The coevolutionary process continued with large deletions in the same regions of the bacterial genomes to block the (E-)PIAS infection. These mutants gained phage resistance via either complete loss or substantial modifications of the phage receptor, MexXY-OprM, negating its essential role in antibiotic resistance. In vitro and in vivo studies indicated that combined use of PIAS and antibiotics could effectively inhibit P. aeruginosa growth. The phage can either eradicate bacteria or induce antibiotic sensitivity in MDR-resistant clinical strains. We have explored the potential use of combination therapy as an alternative approach against MDR P. aeruginosa infection.
外文关键词：bacteriophage therapy；combined treatment；phage resistance；phage-provoked sequential genomic mutation；deletion；MexXY-OprM efflux system
作者：Valappil, SK；Shetty, P；Deim, Z；Terhes, G；Urban, E；Vaczi, S；Patai, R；Polgar, T；Pertics, BZ；Schneider, G；Kovacs, T；Rakhely, G
作者单位：Univ Szeged；Inst Plant Biol；Inst Biophys；Univ Pecs；Enviroinvest Corp；Biopesticide Ltd
期刊名称：FRONTIERS IN MICROBIOLOGY
期刊影响因子：4.236
出版年份：2021
出版刊次：
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