Efflux Pumps of Mycobacterium tuberculosis Play a Significant Role in Antituberculosis Activity of Potential Drug Candidates
01 May 2012-Antimicrobial Agents and Chemotherapy (American Society for Microbiology)-Vol. 56, Iss: 5, pp 2643-2651
TL;DR: It is shown that these four efflux pump KO mutants of M. tuberculosis play a vital role in mediating efflux of different chemical scaffolds and inhibitors of one or several of these efflux pumps could have a significant impact in the treatment of tuberculosis.
Abstract: Active efflux of drugs mediated by efflux pumps that confer drug resistance is one of the mechanisms developed by bacteria to counter the adverse effects of antibiotics and chemicals. To understand these efflux mechanisms in Mycobacterium tuberculosis, we generated knockout (KO) mutants of four efflux pumps of the pathogen belonging to different classes. We measured the MICs and kill values of two different compound classes on the wild type (WT) and the efflux pump (EP) KO mutants in the presence and absence of the efflux inhibitors verapamil and l-phenylalanyl-l-arginyl-β-naphthylamide (PAβN). Among the pumps studied, the efflux pumps belonging to the ABC (ATP-binding cassette) class, encoded by Rv1218c, and the SMR (small multidrug resistance) class, encoded by Rv3065, appear to play important roles in mediating the efflux of different chemical classes and antibiotics. Efflux pumps encoded by Rv0849 and Rv1258c also mediate the efflux of these compounds, but to a lesser extent. Increased killing is observed in WT M. tuberculosis cells by these compounds in the presence of either verapamil or PAβN. The efflux pump KO mutants were more susceptible to these compounds in the presence of efflux inhibitors. We have shown that these four efflux pumps of M. tuberculosis play a vital role in mediating efflux of different chemical scaffolds. Inhibitors of one or several of these efflux pumps could have a significant impact in the treatment of tuberculosis. The identification and characterization of Rv0849, a new efflux pump belonging to the MFS (major facilitator superfamily) class, are reported.
Citations
More filters
01 Jan 2017
TL;DR: The development of clinically-suitable efflux pump inhibitors and novel pump-circumventing antimicrobial drugs continues to be a challenging task and the clinical significance of drug transporters highlights the importance of prudent antimicrobial use for minimizing the emergence and spread of antimicrobial drug resistance.
Abstract: Active drug extrusion mediated by efflux transporters is a phenomenon widely observed in both prokaryotic and eukaryotic cells. This process makes a great contribution to intrinsic and acquired resistance in bacteria, fungi and parasites against a broad range of antimicrobial agents and can further interplay with other resistance mechanisms. Drug transporters also function beyond resistance such as those involved in stress response and virulence. There is an ever-growing understanding of drug transporters with respect to their classification, structure, transport mechanisms, regulation and inhibition. This chapter provides an overview of the characteristics and resistance contributions of drug transporters through various examples of clinical significance, with an emphasis on the transporters of the resistance-nodulation-cell division (RND) superfamily in Gram-negative bacteria. The development of clinically-suitable efflux pump inhibitors and novel pump-circumventing antimicrobial drugs continues to be a challenging task. Additionally, the clinical significance of drug transporters highlights the importance of prudent antimicrobial use for minimizing the emergence and spread of antimicrobial drug resistance.
6 citations
TL;DR: A new medicinal chemistry and biology directed research rational has recently been successfully described in Nature Medicine and will help to fight resistant forms of tuberculosis.
6 citations
TL;DR: In a study that showcases the potential of semisynthetic drug design, structural modification of an existing antibiotic with little activity against Mycobacterium tuberculosis has generated a new class of effective antitubercular lead.
Abstract: In a study that showcases the potential of semisynthetic drug design, structural modification of an existing antibiotic with little activity against Mycobacterium tuberculosis has generated a new class of effective antitubercular lead.
6 citations
TL;DR: DNA repair and damage tolerance as an efficient target for drug development by understanding Mtb DNA repair and tolerance machinery and its regulation, its role in pathogenesis and survival, mutagenesis, and consequently in development of drug resistance is explored.
Abstract: Tuberculosis (TB) is one such disease that has become a nuisance in the world scenario and one of the most deadly diseases of the current times. The etiological agent of tuberculosis, Mycobacterium tuberculosis (M. tb) kills millions of people each year. Not only 1.7 million people worldwide are estimated to harbor M. tb in the latent form but also 5 to 15 percent of which are expected to acquire an infection during a lifetime. Though curable, a long duration of drug regimen and expense leads to low patient adherence. The emergence of multi-, extensive- and total- drug-resistant strains of M. tb further complicates the situation. Owing to high TB burden, scientists worldwide are trying to design novel therapeutics to combat this disease. Therefore, to identify new drug targets, there is a growing interest in targeting DNA repair pathways to fight this infection. Thus, this review aims to explore DNA repair and damage tolerance as an efficient target for drug development by understanding M. tb DNA repair and tolerance machinery and its regulation, its role in pathogenesis and survival, mutagenesis, and consequently, in the development of drug resistance.
6 citations
01 Jan 2016
TL;DR: This chapter provides an overview of molecular and biochemical mechanisms of antimicrobial resistance with an emphasis on the role of drug efflux pumps and their relationship with other key resistance mechanisms in clinically relevant intrinsic and acquired resistance.
Abstract: Antimicrobial agents target several essential cellular functions in bacteria including biosynthesis of the cell wall, nucleic acids, and proteins, which consequently produce inhibitory and even lethal effects on bacterial survival. In contrast, bacterial cells possess remarkable capacities to counteract the action of antimicrobials, thus contributing to resistance. The mechanisms of resistance predominantly involve the production of drug-inactivating enzymes, alteration of drug targets, and prevention of drug access; the latter mechanism refers to the function of drug influx and efflux. This chapter provides an overview of molecular and biochemical mechanisms of antimicrobial resistance with an emphasis on the role of drug efflux pumps and their relationship with other key resistance mechanisms in clinically relevant intrinsic and acquired resistance.
6 citations
References
More filters
TL;DR: Evidence is presented that multidrug-resistance efflux pumps have roles in bacterial pathogenicity and it is proposed that these pumps therefore have greater clinical relevance than is usually attributed to them.
Abstract: It is well established that multidrug-resistance efflux pumps encoded by bacteria can confer clinically relevant resistance to antibiotics. It is now understood that these efflux pumps also have a physiological role(s). They can confer resistance to natural substances produced by the host, including bile, hormones and host-defence molecules. In addition, some efflux pumps of the resistance nodulation division (RND) family have been shown to have a role in the colonization and the persistence of bacteria in the host. Here, I present the accumulating evidence that multidrug-resistance efflux pumps have roles in bacterial pathogenicity and propose that these pumps therefore have greater clinical relevance than is usually attributed to them.
1,367 citations
"Efflux Pumps of Mycobacterium tuber..." refers background in this paper
...tuberculosis within its human host, as well (25)....
[...]
TL;DR: The microbial transcriptome served as a bioprobe of the MTB phagosomal environment, showing it to be nitrosative, oxidative, functionally hypoxic, carbohydrate poor, and capable of perturbing the pathogen's cell envelope.
Abstract: Little is known about the biochemical environment in phagosomes harboring an infectious agent. To assess the state of this organelle we captured the transcriptional responses of Mycobacterium tuberculosis (MTB) in macrophages from wild-type and nitric oxide (NO) synthase 2–deficient mice before and after immunologic activation. The intraphagosomal transcriptome was compared with the transcriptome of MTB in standard broth culture and during growth in diverse conditions designed to simulate features of the phagosomal environment. Genes expressed differentially as a consequence of intraphagosomal residence included an interferon γ– and NO-induced response that intensifies an iron-scavenging program, converts the microbe from aerobic to anaerobic respiration, and induces a dormancy regulon. Induction of genes involved in the activation and β-oxidation of fatty acids indicated that fatty acids furnish carbon and energy. Induction of σE-dependent, sodium dodecyl sulfate–regulated genes and genes involved in mycolic acid modification pointed to damage and repair of the cell envelope. Sentinel genes within the intraphagosomal transcriptome were induced similarly by MTB in the lungs of mice. The microbial transcriptome thus served as a bioprobe of the MTB phagosomal environment, showing it to be nitrosative, oxidative, functionally hypoxic, carbohydrate poor, and capable of perturbing the pathogen's cell envelope.
1,352 citations
"Efflux Pumps of Mycobacterium tuber..." refers background in this paper
...Several efflux pumps and their regulators are also induced during macrophage infection (1, 20, 32, 34)....
[...]
TL;DR: This review focuses on chromosomally encoded pumps in bacteria that cause infections in humans, and suggests that resistance nodulation division systems are important in pathogenicity and/or survival in a particular ecological niche.
Abstract: Efflux pump genes and proteins are present in both antibiotic-susceptible and antibiotic-resistant bacteria. Pumps may be specific for one substrate or may transport a range of structurally dissimilar compounds (including antibiotics of multiple classes); such pumps can be associated with multiple drug (antibiotic) resistance (MDR). However, the clinical relevance of efflux-mediated resistance is species, drug, and infection dependent. This review focuses on chromosomally encoded pumps in bacteria that cause infections in humans. Recent structural data provide valuable insights into the mechanisms of drug transport. MDR efflux pumps contribute to antibiotic resistance in bacteria in several ways: (i) inherent resistance to an entire class of agents, (ii) inherent resistance to specific agents, and (iii) resistance conferred by overexpression of an efflux pump. Enhanced efflux can be mediated by mutations in (i) the local repressor gene, (ii) a global regulatory gene, (iii) the promoter region of the transporter gene, or (iv) insertion elements upstream of the transporter gene. Some data suggest that resistance nodulation division systems are important in pathogenicity and/or survival in a particular ecological niche. Inhibitors of various efflux pump systems have been described; typically these are plant alkaloids, but as yet no product has been marketed.
1,078 citations
"Efflux Pumps of Mycobacterium tuber..." refers background in this paper
...No efflux pump inhibitor has yet reached clinical practice, but it is clear that this area of drug development offers a lot of promise, as it will further enhance the effective use of several drugs that have previously been considered to be of great clinical value and also new molecules that are currently under development (24)....
[...]
...This is possible because of the redundancy of their functions, which may overlap extensively (24, 27)....
[...]
TL;DR: Given the clinical significance of multidrug (and drug-specific) exporters, efflux must be considered in formulating strategies/approaches to treating drug-resistant infections, both in the development of new agents less impacted by efflux and in targeting efflux directly with efflux inhibitors.
Abstract: Antibiotic resistance continues to plague antimicrobial chemotherapy of infectious disease. And while true biocide resistance is as yet unrealized, in vitro and in vivo episodes of reduced biocide susceptibility are common and the history of antibiotic resistance should not be ignored in the development and use of biocidal agents. Efflux mechanisms of resistance, both drug specific and multidrug, are important determinants of intrinsic and/or acquired resistance to these antimicrobials, with some accommodating both antibiotics and biocides. This latter raises the spectre (as yet generally unrealized) of biocide selection of multiple antibiotic-resistant organisms. Multidrug efflux mechanisms are broadly conserved in bacteria, are almost invariably chromosome-encoded and their expression in many instances results from mutations in regulatory genes. In contrast, drug-specific efflux mechanisms are generally encoded by plasmids and/or other mobile genetic elements (transposons, integrons) that carry additional resistance genes, and so their ready acquisition is compounded by their association with multidrug resistance. While there is some support for the latter efflux systems arising from efflux determinants of self-protection in antibiotic-producing Streptomyces spp. and, thus, intended as drug exporters, increasingly, chromosomal multidrug efflux determinants, at least in Gram-negative bacteria, appear not to be intended as drug exporters but as exporters with, perhaps, a variety of other roles in bacterial cells. Still, given the clinical significance of multidrug (and drug-specific) exporters, efflux must be considered in formulating strategies/approaches to treating drug-resistant infections, both in the development of new agents, for example, less impacted by efflux and in targeting efflux directly with efflux inhibitors.
979 citations
"Efflux Pumps of Mycobacterium tuber..." refers background in this paper
...This is possible because of the redundancy of their functions, which may overlap extensively (24, 27)....
[...]
TL;DR: The multifaceted implications of drug efflux transporters warrant novel strategies to combat multidrug resistance in bacteria.
Abstract: Drug efflux pumps play a key role in drug resistance and also serve other functions in bacteria. There has been a growing list of multidrug and drug-specific efflux pumps characterized from bacteria of human, animal, plant and environmental origins. These pumps are mostly encoded on the chromosome, although they can also be plasmid-encoded. A previous article in this journal provided a comprehensive review regarding efflux-mediated drug resistance in bacteria. In the past 5 years, significant progress has been achieved in further understanding of drug resistance-related efflux transporters and this review focuses on the latest studies in this field since 2003. This has been demonstrated in multiple aspects that include but are not limited to: further molecular and biochemical characterization of the known drug efflux pumps and identification of novel drug efflux pumps; structural elucidation of the transport mechanisms of drug transporters; regulatory mechanisms of drug efflux pumps; determining the role of the drug efflux pumps in other functions such as stress responses, virulence and cell communication; and development of efflux pump inhibitors. Overall, the multifaceted implications of drug efflux transporters warrant novel strategies to combat multidrug resistance in bacteria.
755 citations