Treatment of Mycobacterium abscessus(Mab)infections is very challenging due to its intrinsic resistance to most available drugs.Therefore,it is crucial to discover novel anti-Mab drugs.In this study,we explored an int...Treatment of Mycobacterium abscessus(Mab)infections is very challenging due to its intrinsic resistance to most available drugs.Therefore,it is crucial to discover novel anti-Mab drugs.In this study,we explored an intrinsic resistance mechanism through which Mab resists echinomycin(ECH).ECH showed activity against Mab at a minimum inhibitory concentration(MIC)of 2μg/ml.A embC strain in which the embC gene was knocked out showed hypersensitivity to ECH(MIC:0.0078-0.0156μg/ml).The MICs of ECH-resistant strains screened with reference to AembC ranged from 0.25 to 1μg/ml.Mutations in EmbB,including D306A,D306N,R350G,V555l,and G581S,increased the Mab's resistance to ECH when overexpressed in AembC individually(MIC:0.25-0.5μg/ml).These EmbB mutants,edited using the CRISPR/Cpf1 system,showed heightened resistance to ECH(MIC:0.25-0.5μg/ml).The permeability of these Mab strains with edited genes and overexpression was reduced,as evidenced by an ethidium bromide accumulation assay,but it remained significantly higher than that of the parent Mab.In summary,our study demonstrates that ECH exerts potent anti-Mab activity and confirms that EmbB and EmbC are implicated in Mab's sensitivity to ECH.Mutation in EmbB may partially compensate foralossof EmbCfunction.展开更多
基金This work was supported by the National Key R&D Program of China(2021YFA1300900)the National Natural Science Foundation of China(21920102003,82022067,and 22037006)+5 种基金the Chinese Academy of Sciences Grants(154144KYSB20190005 and YJKYYQ20210026)the Key R&D Program of Sichuan Provenience(2023YFSY0047)the State Key Laboratory of Respiratory Disease,Guangzhou Institute of Respiratory Diseases,First Affiliated Hospital of Guangzhou Medical University(SKLRD-Z-202414,SKLRD-OP-202324,SKLRD-Z-202301,SKLRD-OP-202113,and SKLRD-Z-202412)Guangzhou Scienceaand Technology Plan-Youth Doctoral"Sail"Project(2024A04J4273)President's International Fellowship Initiative-CAS(2023VBC0015)the National Foreign Young Talent Program(QN2022031002L).
文摘Treatment of Mycobacterium abscessus(Mab)infections is very challenging due to its intrinsic resistance to most available drugs.Therefore,it is crucial to discover novel anti-Mab drugs.In this study,we explored an intrinsic resistance mechanism through which Mab resists echinomycin(ECH).ECH showed activity against Mab at a minimum inhibitory concentration(MIC)of 2μg/ml.A embC strain in which the embC gene was knocked out showed hypersensitivity to ECH(MIC:0.0078-0.0156μg/ml).The MICs of ECH-resistant strains screened with reference to AembC ranged from 0.25 to 1μg/ml.Mutations in EmbB,including D306A,D306N,R350G,V555l,and G581S,increased the Mab's resistance to ECH when overexpressed in AembC individually(MIC:0.25-0.5μg/ml).These EmbB mutants,edited using the CRISPR/Cpf1 system,showed heightened resistance to ECH(MIC:0.25-0.5μg/ml).The permeability of these Mab strains with edited genes and overexpression was reduced,as evidenced by an ethidium bromide accumulation assay,but it remained significantly higher than that of the parent Mab.In summary,our study demonstrates that ECH exerts potent anti-Mab activity and confirms that EmbB and EmbC are implicated in Mab's sensitivity to ECH.Mutation in EmbB may partially compensate foralossof EmbCfunction.
