Delayed bactericidal response of Mycobacterium tuberculosis to bedaquiline involves remodelling of bacterial metabolism

Koul, Anil; Vranckx, Luc; Dhar, Neeraj; Göhlmann, Hinrich W. H.; Özdemir, Emre; Neefs, Jean-Marc; Schulz, Melanie; Lü, Ping; Mørtz, Ejvind; McKinney, John D.; Andries, Koen; Bald, Dirk

doi:10.1038/ncomms4369

Cited by 240 publications

(301 citation statements)

References 50 publications

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“…In fact, mass spectrometric determination of the metabolome of Mtb-infected human macrophages and granulomatous tissue of guinea pigs show the presence of acetate, lactate, and glutamate (5,54). Moreover, 13 C-metabolic flux analysis of M. tuberculosis grown in THP-1 macrophages showed that glutamate is among the substrates preferentially taken up from the phagosome (55). Although our studies show that PK is not essential for Mtb pathogenesis in mice, it is important to note that mouse models do not perfectly mimic the human disease and that many enzymatic reactions, although non-essential for mouse infection, were shown to be essential to infect higher organisms (56).…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, Mtb maintains a functional and intact glycolytic pathway, suggesting that glycolysis plays a role under certain in vivo conditions. Recent studies with the adenosine triphosphate (ATP) synthase inhibitor, Bedaquiline, showed a delayed cidal effect when Mtb was grown on fermentable carbon sources (13), suggesting that Mtb can produce ATP through substrate level phosphorylation when oxidative-phosphorylation is limited. However, studies in mice with Mtb strains containing knockouts of glycolytic enzymes did not result in strong attenuation, thus leaving the role of glucose metabolism in the virulence and physiology of Mtb unclear (14,15).…”

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confidence: 99%

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Central Role of Pyruvate Kinase in Carbon Co-catabolism of Mycobacterium tuberculosis

Noy¹,

Vergnolle²,

Hartman

et al. 2016

Journal of Biological Chemistry

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Mycobacterium tuberculosis (Mtb) displays a high degree of metabolic plasticity to adapt to challenging host environments. Genetic evidence suggests that Mtb relies mainly on fatty acid catabolism in the host. However, Mtb also maintains a functional glycolytic pathway and its role in the cellular metabolism of Mtb has yet to be understood. Pyruvate kinase catalyzes the last and rate-limiting step in glycolysis and the Mtb genome harbors one putative pyruvate kinase (pykA, Rv1617). Here we show that pykA encodes an active pyruvate kinase that is allosterically activated by glucose 6-phosphate (Glc-6-P) and adenosine monophosphate (AMP). Deletion of pykA prevents Mtb growth in the presence of fermentable carbon sources and has a cidal effect in the presence of glucose that correlates with elevated levels of the toxic catabolite methylglyoxal. Growth attenuation was also observed in media containing a combination of short chain fatty acids and glucose and surprisingly, in media containing odd and even chain fatty acids alone. Untargeted high sensitivity metabolomics revealed that inactivation of pyruvate kinase leads to accumulation of phosphoenolpyruvate (P-enolpyruvate), citrate, and aconitate, which was consistent with allosteric inhibition of isocitrate dehydrogenase by P-enolpyruvate. This metabolic block could be relieved by addition of the ␣-ketoglutarate precursor glutamate. Taken together, our study identifies an essential role of pyruvate kinase in preventing metabolic block during carbon co-catabolism in Mtb.Mycobacterium tuberculosis (Mtb) 3 pathogenesis has been studied for decades, however, our knowledge concerning the metabolism and physiology of the bacterium during host infection is still limited (1-4). Specifically, we lack understanding of nutrient availability in the host microenvironments during the different phases of infection and consequently, which nutrients (e.g. carbon and nitrogen sources) are used by the bacterium for growth and maintenance of replicative and non-replicative states (5). In vitro evidence suggests that Mtb does not utilize carbon catabolite repression, a regulatory mechanism that allows bacteria and single-cell eukaryotes to gain growth advantage through prioritized metabolism of one carbon source over the other (6), but rather co-catabolizes multiple carbon sources at once (4). Several lines of evidence suggest that Mtb relies mainly on fatty acid metabolism in the non-replicative state within the host (1, 7-12). During growth on fatty acids Mtb bypasses the carbon dioxide releasing steps of the TCA cycle by running the glyoxylate shunt to conserve carbon (4). Nevertheless, Mtb maintains a functional and intact glycolytic pathway, suggesting that glycolysis plays a role under certain in vivo conditions. Recent studies with the adenosine triphosphate (ATP) synthase inhibitor, Bedaquiline, showed a delayed cidal effect when Mtb was grown on fermentable carbon sources (13), suggesting that Mtb can produce ATP through substrate level phosphorylation when oxidative-phosph...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Central Role of Pyruvate Kinase in Carbon Co-catabolism of Mycobacterium tuberculosis

Noy¹,

Vergnolle²,

Hartman

et al. 2016

Journal of Biological Chemistry

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“…Like clofazimine, the half-life of bedaquiline is extremely long (days to weeks) and is dependent on the duration of administration (31,32), and bedaquiline is also highly protein bound in the serum (30,31), with free drug concentration estimates well below the MIC of the drug for M. tuberculosis. Bedaquiline also exhibits delayed bactericidal activity against M. tuberculosis, both in vitro (33,34) and in vivo (32,35), and it has been hypothesized that this delay is related to bedaquiline's mechanism of action targeting ATP synthase (36), with bactericidal activity evident only after bacterial energy stores have been depleted (33). Finally, low-level, efflux-driven cross-resistance to bedaquiline and clofazimine has been reported in M. tuberculosis (37).…”

Section: Discussionmentioning

confidence: 99%

Pharmacokinetics and Pharmacodynamics of Clofazimine in a Mouse Model of Tuberculosis

Swanson

Adamson

Moodley

et al. 2015

Antimicrob Agents Chemother

115

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fThe antileprosy drug clofazimine has shown potential for shortening tuberculosis treatment; however, the current dosing of the drug is not evidence based, and the optimal dosing is unknown. Our objective was to conduct a preclinical evaluation of the pharmacokinetics and pharmacodynamics of clofazimine in the mouse model of tuberculosis, with the goal of providing useful information on dosing for future studies. Pharmacokinetic parameters were evaluated in infected and uninfected BALB/c mice. Pharmacodynamic parameters were evaluated in Mycobacterium tuberculosis-infected mice that were treated for 12 weeks with one of six different clofazimine dosing regimens, i.e., doses of 6.25, 12.5, and 25 mg/kg of body weight/day and 3 regimens with loading doses. Clofazimine progressively accumulated in the lungs, livers, and spleens of the mice, reaching levels of greater than 50 g/g in all tissues by 4 weeks of administration, while serum drug levels remained low at 1 to 2 g/ml. Elimination of clofazimine was extremely slow, and the half-life was dependent on the duration of drug administration. Clofazimine exhibited dosedependent tissue and serum concentrations. At any dose, clofazimine did not have bactericidal activity during the first 2 weeks of administration but subsequently demonstrated potent, dose-independent bactericidal activity. The antituberculosis activity of clofazimine was dependent on neither the dose administered nor the drug concentrations in the tissues, suggesting that much lower doses could be effectively used for tuberculosis treatment. C lofazimine (CFZ) is a phenazine dye developed in the 1950s for the treatment of tuberculosis (TB) (1, 2). Despite promising activity against Mycobacterium tuberculosis both in vitro and in vivo, clofazimine was not advanced as a TB drug but instead became incorporated decades later into the multidrug treatment of leprosy, where it remains a key drug (3, 4). Interest in clofazimine for TB treatment has been revitalized following the 2010 report by Van Deun and colleagues that a clofazimine-containing regimen not only was highly effective for the treatment of multidrug-resistant (MDR) TB, but was also associated with a significantly decreased duration of therapy (5); 9 months of a clofazimine-containing regimen resulted in 87.9% relapse-free cure, which is in sharp contrast to the limited efficacy (less than 50% relapse-free cure) of the World Health Organization (WHO)-recommended, at least 20-month-long MDR-TB treatment regimen (6, 7). This report was complemented by experimental chemotherapy studies in which the inclusion of clofazimine in first-and secondline regimens significantly shortened the duration of treatment needed for relapse-free cure in mouse models of drug-susceptible and MDR TB, respectively (8, 9). These clinical and preclinical data suggest that clofazimine has great potential for TB treatment.Because clofazimine was abandoned as an option for TB treatment shortly after its discovery, and also because the use of clofazimine for leprosy is not ba...

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“…Remarkably, BDQ was also discovered to also show activity against hypoxic, nonreplicating Mtb (Rao et al 2008). Apart from its potency, BDQ was also found to show an unusually slow time-dependent killing such that only bacteriostatic activity was observed during the first 3-4 d but was then followed by a bactericidal phase with a 4 log 10 reduction in measurable colony-forming units by day 14 (Koul et al 2014).…”

Section: Bedaquiline (Bdq)mentioning

confidence: 96%

Tuberculosis Drug Development: History and Evolution of the Mechanism-Based Paradigm : Figure 1.

Chakraborty

Rhee

2015

Cold Spring Harb Perspect Med

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Modern tuberculosis (TB) chemotherapy is widely viewed as a crowning triumph of antiinfectives research. However, only one new TB drug has entered clinical practice in the past 40 years while drug resistance threatens to further destabilize the pandemic. Here, we review a brief history of TB drug development, focusing on the evolution of mechanism(s)-of-action studies and key conceptual barriers to rational, mechanism-based drugs.

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Delayed bactericidal response of Mycobacterium tuberculosis to bedaquiline involves remodelling of bacterial metabolism

Cited by 240 publications

References 50 publications

Central Role of Pyruvate Kinase in Carbon Co-catabolism of Mycobacterium tuberculosis

Central Role of Pyruvate Kinase in Carbon Co-catabolism of Mycobacterium tuberculosis

Pharmacokinetics and Pharmacodynamics of Clofazimine in a Mouse Model of Tuberculosis

Tuberculosis Drug Development: History and Evolution of the Mechanism-Based Paradigm : Figure 1.

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