Abstract
Drug-resistant tuberculosis (TB) is a growing public health problem. There is an urgent need for information regarding cross-resistance and collateral sensitivity relationships among drugs and the genetic determinants of anti-TB drug resistance for developing strategies to suppress the emergence of drug-resistant pathogens. To identify mutations that confer resistance to anti-TB drugs in Mycobacterium species, we performed the laboratory evolution of nonpathogenic Mycobacterium smegmatis, which is closely related to Mycobacterium tuberculosis, against ten anti-TB drugs. Next, we performed whole-genome sequencing and quantified the resistance profiles of each drug-resistant strain against 24 drugs. We identified the genes with novel meropenem (MP) and linezolid (LZD) resistance-conferring mutation, which also have orthologs, in M. tuberculosis H37Rv. Among the 240 possible drug combinations, we identified 24 pairs that confer cross-resistance and 18 pairs that confer collateral sensitivity. The acquisition of bedaquiline or linezolid resistance resulted in collateral sensitivity to several drugs, while the acquisition of MP resistance led to multidrug resistance. The MP-evolved strains showed cross-resistance to rifampicin and clarithromycin owing to the acquisition of a mutation in the intergenic region of the Rv2864c ortholog, which encodes a penicillin-binding protein, at an early stage. These results provide a new insight to tackle drug-resistant TB.
Highlights
Tuberculosis (TB) is one of the top ten causes of death, exceeding HIV as the leading cause of death from a single infectious agent[1]
We showed that the acquisition of Meropenem (MP) resistance owing to the newly identified mutation in the gene encoding penicillin-binding protein (PBP) confers multidrug-resistance to the strain
Since M. smegmatis forms aggregates in liquid media, we developed a new method for laboratory evolution on an agar plate under a drug gradient
Summary
Tuberculosis (TB) is one of the top ten causes of death, exceeding HIV as the leading cause of death from a single infectious agent[1]. Antibiotic pairs that exhibit collateral sensitivities and the corresponding genetic determinants have been studied extensively through parallel laboratory evolution in whole-genome sequencing and phenotyping assays[14,16,17,18,19,20] These studies identified novel collateral-sensitivity and cross-resistance interactions and proposed new, rational treatment strategies to exploit collateral sensitivity[9,17]. We showed that the acquisition of Meropenem (MP) resistance owing to the newly identified mutation in the gene encoding penicillin-binding protein (PBP) confers multidrug-resistance to the strain This is the first study to report the comprehensive laboratory evolution of Mycobacterium with respect to anti-TB drug resistance
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