Two New Dihydrosphingosine Analogs Against Mycobacterium tuberculosis Affect gltA1, lprQ, and rpsO Expression.

Katia Peñuelas-Urquides,Jorge Castro-Garza,Salvador Said-Fernández,Pola Becerril-Montes,Mario Bermúdez De León,Arturo San Feliciano,Fabiola Castorena-Torres,Beatriz Silva-Ramírez,Laura Adiene González-Escalante,Licet Villarreal-Treviño,Gloria María Molina-Salinas,Esther Del Olmo

doi:10.3389/fmicb.2021.742867

Abstract

The emergence of multidrug-resistant (MDR) Mycobacterium tuberculosis strains threaten the control of tuberculosis. New antitubercular dihydrosphingosine analogs, named UCIs, have been evaluated in preclinical studies but their cellular and molecular mechanisms of action against M. tuberculosis are still unknown. The aim of this study was to evaluate the effect of UCI exposure on gene expression of drug-sensitive H37Rv and MDR CIBIN:UMF:15:99 clones of M. tuberculosis which were isolated, phenotypically, and genetically characterized, cultured to log phase and treated with UCI compounds; followed by total RNA isolation, reverse transcription and hybridization assays on Affymetrix genomic microarrays. Data were validated with RT-qPCR assays. As results, UCI-05 and UCI-14 exposure increased gltA1 expression in drug-sensitive H37Rv clones. Furthermore, UCI-05 increased lprQ expression in MDR CIBIN:UMF:15:99 M. tuberculosis clones while UCI-14 reduced the expression of this gene in drug-sensitive H37Rv clones. In addition, UCI-05 reduced rpsO expression in drug-sensitive H37Rv clones. We found gene expression alterations that suggest these molecules may alter carbon and lipid metabolism as well as interfere in the protein-producing machinery in M. tuberculosis.

Highlights

Tuberculosis (TB) is caused primarily by Mycobacterium tuberculosis and is one of the leading causes of death attributable to a single infectious agent
M. tuberculosis clones were derived from the H37Rv strain (ATCC 27294) and from the clinical isolate CIBIN:UMF:15:99, which is resistant to all basic antitubercular medications [streptomycin (STR), isoniazid (INH), rifampicin (RIF), ethambutol (EMB), and pyrazinamide (PZA)] (Molina-Salinas et al, 2006)
M. tuberculosis clones were derived from the H37Rv strain (ATCC 27294), which is sensitive to first-line antitubercular drugs, and from the MDR clinical isolate- CIBIN:UMF:15:99

Summary

Introduction

Tuberculosis (TB) is caused primarily by Mycobacterium tuberculosis and is one of the leading causes of death attributable to a single infectious agent. The development of new antitubercular drugs is essential to limit morbidity, mortality and transmission of these strains. UCI-05 and UCI-14 (Figure 1) have shown good in vitro and in vivo activity against first line antitubercular drug susceptible and MDR strains, with minimal acute and subchronic toxicity in BALB/c mice (PCT EP 2007062381, WO 2008059014 A2) (Del Olmo et al, 2009; Olmo et al, 2016). The study of drug-induced changes of gene expression profiles at the transcriptional level may generate new insights regarding genes related to the mechanism of action (Fu, 2006). We aimed to identify those genes modified by UCI-05 and UCI-14 exposure to acquire relevant information in the mechanism of action of these compounds in drug-sensitive and MDR strains of M. tuberculosis

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