Mycobacterium abscessus Genetic Determinants Associated with the Intrinsic Resistance to Antibiotics

Mylene Gorzynski; Tiana Week; Tiana Jaramillo; Elizaveta Dzalamidze; Lia Danelishvili

doi:10.3390/microorganisms9122527

Mycobacterium abscessus Genetic Determinants Associated with the Intrinsic Resistance to Antibiotics

Microorganisms. 2021 Dec 7;9(12):2527. doi: 10.3390/microorganisms9122527.

Authors

Mylene Gorzynski^{1

2}, Tiana Week^{1

3}, Tiana Jaramillo^{1

4}, Elizaveta Dzalamidze^{1

5}, Lia Danelishvili¹

Affiliations

¹ Department of Biomedical Sciences, Carlson College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA.
² Department of Biochemistry & Molecular Biology, Oregon State University, Corvallis, OR 97331, USA.
³ Department of Bioengineering, College of Engineering, Oregon State University, Corvallis, OR 97331, USA.
⁴ Department of Animal Sciences, College of Agricultural Sciences, Oregon State University, Corvallis, OR 97331, USA.
⁵ BioHealth Sciences, Department of Microbiology, College of Sciences, Oregon State University, Corvallis, OR 97331, USA.

Abstract

Mycobacterium abscessus subsp. abscessus (MAB) is a fast-growing nontuberculous mycobacterium causing pulmonary infections in immunocompromised and immunocompetent individuals. The treatment of MAB infections in clinics is extremely challenging, as this organism is naturally resistant to most available antibiotics. There is limited knowledge on the mechanisms of MAB intrinsic resistance and on the genes that are involved in the tolerance to antimicrobials. To identify the MAB genetic factors, including the components of the cell surface transport systems related to the efflux pumps, major known elements contributing to antibiotic resistance, we screened the MAB transposon library of 2000 gene knockout mutants. The library was exposed at either minimal inhibitory (MIC) or bactericidal concentrations (BC) of amikacin, clarithromycin, or cefoxitin, and MAB susceptibility was determined through the optical density. The 98 susceptible and 36 resistant mutants that exhibited sensitivity below the MIC and resistance to BC, respectively, to all three drugs were sequenced, and 16 mutants were found to belong to surface transport systems, such as the efflux pumps, porins, and carrier membrane enzymes associated with different types of molecule transport. To establish the relevance of the identified transport systems to antibiotic tolerance, the gene expression levels of the export related genes were evaluated in nine MAB clinical isolates in the presence or absence of antibiotics. The selected mutants were also evaluated for their ability to form biofilms and for their intracellular survival in human macrophages. In this study, we identified numerous MAB genes that play an important role in the intrinsic mechanisms to antimicrobials and further demonstrated that, by targeting components of the drug efflux system, we can significantly increase the efficacy of the current antibiotics.

Keywords: M. abscessus; antibiotic treatment; biofilm; efflux pumps; intrinsic resistance; macrophage; rapidly growing mycobacteria; virulence factors.

Grants and funding

VMDGDR-VG78/OSU Biomedical Department Research Grant