Elucidation of Mycobacterium abscessus aminoglycoside and capreomycin resistance by targeted deletion of three putative resistance genes

Objectives: Mycobacterium abscessus is innately resistant to a variety of drugs thereby limiting therapeutic options. Bacterial resistance to aminoglycosides (AGs) is conferred mainly by AG-modifying enzymes, which often have overlapping activities. Several putative AG-modifying enzymes are encoded in the genome of M. abscessus. The aim of this study was to investigate the molecular basis underlying AG resistance in M. abscessus. Methods: M. abscessus deletion mutants deficient in one of three genes potentially involved in AG resistance, aac(2ʹ), eis1 and eis2, were generated by targeted gene inactivation, as were combinatorial double and triple deletion mutants. MICs were determined to study susceptibility to a variety of AG drugs and to capreomycin. Results: Deletion of aac(2ʹ) increased susceptibility of M. abscessus to kanamycin B, tobramycin, dibekacin and gentamicin C. Deletion of eis2 increased susceptibility to capreomycin, hygromycin B, amikacin and kanamycin B. Deletion of eis1 did not affect drug susceptibility. Equally low MICs of apramycin, arbekacin, isepamicin and kanamycin A for WT and mutant strains indicate that these drugs are not inactivated by either AAC(2ʹ) or Eis enzymes. Conclusions: M. abscessus expresses two distinct AG resistance determinants, AAC(2ʹ) and Eis2, which confer clinically relevant drug resistance