Isoniazid and verapamil modulatory activity and efflux pump gene expression in <i>Mycobacterium tuberculosis</i>

Souza, Joao V. P. de; Murase, Letícia Sayuri; Caleffi-Ferracioli, Katiany Rizzieri; Palomo, Carolina Trevisolli; Scodro, Regiane Bertin de Lima; Siqueira, Vera Ld; Pavan, Fernando Rogério; Cardoso, Rosilene Fressatti

doi:10.5588/ijtld.19.0458

Cited by 5 publications

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“…In some previous studies, it was noted that when VP was used in combination with anti-tuberculosis drugs [including first- and second-line drugs ( 11 , 29 , 30 ) as well as some newly developed new drugs ( 31 )], it showed significant effects both in vivo and in vitro . However, in this study, VP did not have a significant effect on INH mono-resistant isolates, which is consistent with the results of previous studies ( 17 ). However, in contrast, other studies ( 12 , 17 ) pointed out that VP reduces the INH MICs for almost all INH-resistant strains.…”

Section: Discussionsupporting

confidence: 93%

“…However, in this study, VP did not have a significant effect on INH mono-resistant isolates, which is consistent with the results of previous studies ( 17 ). However, in contrast, other studies ( 12 , 17 ) pointed out that VP reduces the INH MICs for almost all INH-resistant strains. Possible explanations for such contradictory results are that the efflux pump genes did not show significant overexpression in the clinical isolates in this study, so that VP did not show significant effect to reverse bacterial resistance.…”

Section: Discussionsupporting

confidence: 93%

“…These molecules can inhibit the efflux of anti-tuberculosis drugs, improve drug efficacy, reverse resistance, and create synergistic effects when used with first-line anti-tuberculosis drugs. Verapamil has been shown to effectively inhibit drug efflux in previous studies ( 16 , 17 ), and studies have demonstrated that the combination of verapamil with isoniazid or rifampicin (RIF) can reduce the MIC of INH or RIF and even reverse the resistance of Mtb.…”

Section: Introductionmentioning

confidence: 99%

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Overexpression of efflux pump genes is one of the mechanisms causing drug resistance in Mycobacterium tuberculosis

Long,

Wang,

Xie

et al. 2024

Microbiol Spectr

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Currently, drug resistance is a formidable obstacle to tuberculosisprevention globally. A deep understanding of the drug-resistance mechanism of Mycobacterium tuberculosis is helpful to deal with this problem. In this study, 46 clinical isolates of Mycobacterium tuberculosis from Zigong, Sichuan, China, including 5 sensitive isolates, 5 rifampicin (RIF) mono-resistant isolates, 18 isoniazid mono-resistant isolates, and 18 multi-drug-resistant isolates, were selected. We evaluated the impact of drug induction on the relative expression levels of 10 putative efflux pump genes, namely, Rv3065 , Rv2836c , efpA , Rv1410c , Rv1250 , Rv0876c , Rv1819c , Rv0933 , Rv1217c , and Rv1218c . Meanwhile, we assessed the effect of verapamil, an efflux pump inhibitor, on the drug resistance of the isolates. The results showed that 100% (5/5) of the RIF mono-resistant isolates, 44.4% (8 of 18) of the isoniazid mono-resistant isolates, and 88.9% (16 of 8) of the multi-drug-resistant isolates overexpressed anyone of the 10 efflux pump genes; however, none of the 10 efflux pump genes were overexpressed on the sensitive isolates. Among drug-resistant strains, Rv1250 (51.2%) and Rv0933 (53.7%) were the genes with the highest frequency of overexpression. Only the rifampicin mono-resistant isolates showed a significant increase in the number of overexpressed efflux pump genes after drug induction, while the change trends of the numbers of overexpressed efflux pump genes in isoniazid mono-resistant isolates and multi-drug-resistant isolates after drug induction were not definite. Verapamil did not have a significant effect on the minimum inhibitory concentrations (MICs) of isoniazid mono-resistant isolates but reduced the MICs of all sensitive isolates and some multi-drug-resistant isolates. We also investigated the correlation between polymorphic loci of mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing, mutations in drug resistance-associated genes, and overexpression of the efflux pump genes. The results indicated that there was no significant association between the two most polymorphic loci, MIRU26 and MIRU31, and the incidence of overexpression of the efflux pump genes. However, isolates with mutations in drug resistance-associated genes exhibited a higher rate of overexpression of the efflux pump genes compared to those without mutations. In conclusion, we found that drug-resistant isolates had a higher background expression level of the efflux pump genes compared to sensitive isolates. This suggests that the overexpression of the efflux pump genes could be one of the mechanisms causing drug resistance in Mycobacterium tuberculosis . We also found that verapamil was able to reduce MICs in some drug-resistant isolates, indicating that inhibition of the efflux pump could be used as an adjuvant treatment option for drug-resistant tuberculosis patients. IMPORTANCE Gene mutations cannot explain all drug resistance of Mycobacterium tuberculosis , and the overexpression of efflux pump genes is considered another important cause of drug resistance. A total of 46 clinical isolates were included in this study to analyze the overexpression of efflux pump genes in different resistant types of strains. The results showed that overexpression of efflux pump genes did not occur in sensitive strains. There was no significant trend in the overexpression of efflux pump genes before and after one-half of MIC drug induction. By adding the efflux pump inhibitor verapamil, we can observe the decrease of MIC of some drug-resistant strains. At the same time, this study ensured the reliability of calculating the relative expression level of efflux pump genes by screening reference genes and using two reference genes for the normalization of quantitative PCR. Therefore, this study confirms that the overexpression of efflux pump genes plays an important role in the drug resistance of clinical isolates of Mycobacterium tuberculosis .

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Overexpression of efflux pump genes is one of the mechanisms causing drug resistance in Mycobacterium tuberculosis

Long,

Wang,

Xie

et al. 2024

Microbiol Spectr

View full text Add to dashboard Cite

show abstract

“… 36 Previous studies showed verapamil, one of the efflux pump inhibitors, could effectively inhibit drug efflux. 37 At the same time, studies have also demonstrated that combining verapamil with isoniazid or rifampicin (RIF) can cause the MIC of isoniazid (INH) or RIF reduction and even reverse the resistance of MTB. 38 About 20 genes in the genus MTB have been identified as related genes encoding efflux pumps, mainly concentrated in MFS, RND, and SMR.…”

Section: Antibiotic Resistance Of Mtbmentioning

confidence: 99%

Identification of Mycobacterium tuberculosis Resistance to Common Antibiotics: An Overview of Current Methods and Techniques

Xiong,

Zhang,

Yan

et al. 2024

IDR

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Multidrug-resistant tuberculosis (MDR-TB) is an essential cause of tuberculosis treatment failure and death of tuberculosis patients. The rapid and reliable profiling of Mycobacterium tuberculosis (MTB) drug resistance in the early stage is a critical research area for public health. Then, most traditional approaches for detecting MTB are time-consuming and costly, leading to the inappropriate therapeutic schedule resting on the ambiguous information of MTB drug resistance, increasing patient economic burden, morbidity, and mortality. Therefore, novel diagnosis methods are frequently required to meet the emerging challenges of MTB drug resistance distinguish. Considering the difficulty in treating MDR-TB, it is urgently required for the development of rapid and accurate methods in the identification of drug resistance profiles of MTB in clinical diagnosis. This review discussed recent advances in MTB drug resistance detection, focusing on developing emerging approaches and their applications in tangled clinical situations. In particular, a brief overview of antibiotic resistance to MTB was present, referred to as intrinsic bacterial resistance, consisting of cell wall barriers and efflux pumping action and acquired resistance caused by genetic mutations. Then, different drug susceptibility test (DST) methods were described, including phenotype DST, genotype DST and novel DST methods. The phenotype DST includes nitrate reductase assay, Roche TM solid ratio method, and liquid culture method and genotype DST includes fluorescent PCR, GeneXpert, PCR reverse dot hybridization, ddPCR, next-generation sequencing and gene chips. Then, novel DST methods were described, including metabolism testing, cell-free DNA probe, CRISPR assay, and spectral analysis technique. The limitations, challenges, and perspectives of different techniques for drug resistance are also discussed. These methods significantly improve the detection sensitivity and accuracy of multidrug-resistant tuberculosis (MRT) and can effectively curb the incidence of drug-resistant tuberculosis and accelerate the process of tuberculosis eradication.

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“…Additionally, LTCC ion channel blockers can alter iron-associated metabolic pathways thereby impeding iron accessibility which lowers intracellular bacterial survival ( 143 ). Furthermore, verapamil acts synergistically with first-line anti-TB drugs- INH ( 152 ), and RIF ( 153 ) in lowering the bacterial burden in cultures, macrophages, and murine models of TB. In another study, antimycobacterial activity of verapamil was confirmed with several TB drugs including bedaquiline (BDQ) and clofazimine (CFZ) and decreased bacterial load was linked with induction of membrane stress responses as a consequence of verapamil induced membrane function disruption ( 154 ).…”

Section: Targeting Host Ion Channelsmentioning

confidence: 99%

Progressive Host-Directed Strategies to Potentiate BCG Vaccination Against Tuberculosis

2022

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The pursuit to improve the TB control program comprising one approved vaccine, M. bovis Bacille Calmette-Guerin (BCG) has directed researchers to explore progressive approaches to halt the eternal TB pandemic. Mycobacterium tuberculosis (M.tb) was first identified as the causative agent of TB in 1882 by Dr. Robert Koch. However, TB has plagued living beings since ancient times and continues to endure as an eternal scourge ravaging even with existing chemoprophylaxis and preventive therapy. We have scientifically come a long way since then, but despite accessibility to the standard antimycobacterial antibiotics and prophylactic vaccine, almost one-fourth of humankind is infected latently with M.tb. Existing therapeutics fail to control TB, due to the upsurge of drug-resistant strains and increasing incidents of co-infections in immune-compromised individuals. Unresponsiveness to established antibiotics leaves patients with no therapeutic possibilities. Hence the search for an efficacious TB immunization strategy is a global health priority. Researchers are paving the course for efficient vaccination strategies with the radically advanced operation of core principles of protective immune responses against M.tb. In this review; we have reassessed the progression of the TB vaccination program comprising BCG immunization in children and potential stratagems to reinforce BCG-induced protection in adults.

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Isoniazid and verapamil modulatory activity and efflux pump gene expression in Mycobacterium tuberculosis

Cited by 5 publications

References 0 publications

Overexpression of efflux pump genes is one of the mechanisms causing drug resistance in Mycobacterium tuberculosis

Overexpression of efflux pump genes is one of the mechanisms causing drug resistance in Mycobacterium tuberculosis

Identification of Mycobacterium tuberculosis Resistance to Common Antibiotics: An Overview of Current Methods and Techniques

Progressive Host-Directed Strategies to Potentiate BCG Vaccination Against Tuberculosis

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