High-dose rifampicin kills persisters, shortens treatment duration, and reduces relapse rate in vitro and in vivo
Yanmin Hu, Alexander Liu, Fatima Ortega-Muro, Laura Alameda-Martin, Denis Mitchison, Anthony Coates, Yanmin Hu, Alexander Liu, Fatima Ortega-Muro, Laura Alameda-Martin, Denis Mitchison, Anthony Coates
Abstract
Although high-dose rifampicin holds promise for improving tuberculosis control by potentially shortening treatment duration, these effects attributed to eradication of persistent bacteria are unclear. The presence of persistent Mycobacterium tuberculosis was examined using resuscitation promoting factors (RPFs) in both in vitro hypoxia and in vivo murine tuberculosis models before and after treatment with incremental doses of rifampicin. Pharmacokinetic parameters and dose-dependent profile of rifampicin in the murine model were determined. The Cornell mouse model was used to test efficacy of high-dose rifampicin in combination with isoniazid and pyrazinamide and to measure relapse rate. There were large numbers of RPF-dependent persisters in vitro and in vivo. Stationary phase cultures were tolerant to rifampicin while higher concentrations of rifampicin eradicated plate count positive but not RPF-dependent persistent bacteria. In murine infection model, incremental doses of rifampicin exhibited a dose-dependent eradication of RPF-dependent persisters. Increasing the dose of rifampicin significantly reduced the risk of antibiotic resistance emergence. In Cornell model, mice treated with high-dose rifampicin regimen resulted in faster visceral clearance; organs were M. tuberculosis free 8 weeks post-treatment compared to 14 weeks with standard-dose rifampicin regimen. Organ sterility, plate count and RPF-dependent persister negative, was achieved. There was no disease relapse compared to the standard dose regimen (87.5%). High-dose rifampicin therapy results in eradication of RPF-dependent persisters, allowing shorter treatment duration without disease relapse. Optimizing rifampicin to its maximal efficacy with acceptable side-effect profiles will provide valuable information in human studies and can potentially improve current tuberculosis chemotherapy.
Keywords: Mycobacterium tuberculosis; mouse model; persistence; resuscitation promoting factors; rifampicin.
Figures
References
- Abdool Karim S. S., Naidoo K., Grobler A., Padayatchi N., Baxter C., Gray A., et al. (2010). Timing of initiation of antiretroviral drugs during tuberculosis therapy. N. Engl. J. Med. 362 697–706. 10.1056/NEJMoa0905848
- Boeree M. J., Diacon A., Dawson R., Van Balen G. P., Venter A., Du Bois J., et al. (2013). What is the Right Dose of Rifampin? A Dose Escalating Study. Available at:
- Calver A. D., Falmer A. A., Murray M., Strauss O. J., Streicher E. M., Hanekom M., et al. (2010). Emergence of increased resistance and extensively drug-resistant tuberculosis despite treatment adherence, South Africa. Emerg. Infect. Dis. 16 264–271. 10.3201/eid1602.090968
- Coates A. R., Hu Y., Jindani A., Mitchison D. A. (2013). Contradictory results with high-dosage rifamycin in mice and humans. Antimicrob. Agents Chemother. 57 1103 10.1128/AAC.01705-12
- de Steenwinkel J. E., Aarnoutse R. E., de Knegt G. J., Ten Kate M. T., Teulen M., Verbrugh H. A., et al. (2013). Optimization of the rifampin dosage to improve the therapeutic efficacy in tuberculosis treatment, using a murine model. Am. J. Respir. Crit. Care Med. 187 1127–1134. 10.1164/rccm.201207-1210OC
- de Steenwinkel J. E., de Knegt G. J., ten Kate M. T., van Belkum A., Verbrugh H. A., Kremer K., et al. (2010). Time-kill kinetics of anti-tuberculosis drugs, and emergence of resistance, in relation to metabolic activity of Mycobacterium tuberculosis. J. Antimicrob. Chemother. 65 2582–2589. 10.1093/jac/dkq374
- Diacon A. H., Patientia R. F., Venter A., van Helden P. D., Smith P. J., McIlleron H., et al. (2007). Early bactericidal activity of high-dose rifampin in patients with pulmonary tuberculosis evidenced by positive sputum smears. Antimicrob. Agents Chemother. 51 2994–2996. 10.1128/AAC.01474-06
- Doble N., Shaw R., Rowland-Hill C., Lush M., Warnock D. W., Keal E. E. (1988). Pharmacokinetic study of the interaction between rifampicin and ketoconazole. J. Antimicrob. Chemother. 21 633–635. 10.1093/jac/21.5.633
- Gill W. P., Harik N. S., Whiddon M. R., Liao R. P., Mittler J. E., Sherman D. R. (2009). A replication clock for Mycobacterium tuberculosis. Nat. Med. 15 211–214. 10.1038/nm.1915
- Girling D. J. (1978). The hepatic toxicity of antituberculosis regimens containing isoniazid, rifampicin and pyrazinamide. Tubercle 59 13–32. 10.1016/0041-3879(77)90022-8
- Gumbo T., Louie A., Deziel M. R., Liu W., Parsons L. M., Salfinger M., et al. (2007). Concentration-dependent Mycobacterium tuberculosis killing and prevention of resistance by rifampin. Antimicrob. Agents Chemother. 51 3781–3788. 10.1128/AAC.01533-06
- Hu Y., Butcher P. D., Mangan J. A., Rajandream M. A., Coates A. R. (1999). Regulation of hmp gene transcription in Mycobacterium tuberculosis: effects of oxygen limitation and nitrosative and oxidative stress. J. Bacteriol. 181 3486–3493.
- Hu Y. M., Butcher P. D., Sole K., Mitchison D. A., Coates A. R. (1998). Protein synthesis is shutdown in dormant Mycobacterium tuberculosis and is reversed by oxygen or heat shock. FEMS Microbiol. Lett. 158 139–145. 10.1111/j.1574-6968.1998.tb12813.x
- Hu Y., Coates A. R. (2009). Acute and persistent Mycobacterium tuberculosis infections depend on the thiol peroxidase TPX. PLoS ONE 4:e5150 10.1371/journal.pone.0005150
- Hu Y., Mangan J. A., Dhillon J., Sole K. M., Mitchison D. A., Butcher P. D., et al. (2000). Detection of mRNA transcripts and active transcription in persistent Mycobacterium tuberculosis induced by exposure to rifampin or pyrazinamide. J. Bacteriol. 182 6358–6365. 10.1128/JB.182.22.6358-6365.2000
- Hu Y., Movahedzadeh F., Stoker N. G., Coates A. R. (2006). Deletion of the Mycobacterium tuberculosis alpha-crystallin-like hspX gene causes increased bacterial growth in vivo. Infect. Immun. 74 861–868. 10.1128/IAI.74.2.861-868.2006
- Jayaram R., Gaonkar S., Kaur P., Suresh B. L., Mahesh B. N., Jayashree R., et al. (2003). Pharmacokinetics-pharmacodynamics of rifampin in an aerosol infection model of tuberculosis. Antimicrob. Agents Chemother. 47 2118–2124. 10.1128/AAC.47.7.2118-2124.2003
- Kreis B., Pretet S., Birenbaum J., Guibout P., Hazeman J. J., Orin E., et al. (1976). Two three-month treatment regimens for pulmonary tuberculosis. Bull. Int. Union Tuberc. 51 71–75.
- McCune R. M., Feldmann F. M., Lambert H. P., McDermott W. (1966). Microbial persistence. I. The capacity of tubercle bacilli to survive sterilization in mouse tissues. J. Exp. Med. 123 445–468. 10.1084/jem.123.3.445
- McCune R. M., Jr., Tompsett R. (1956). Fate of Mycobacterium tuberculosis in mouse tissues as determined by the microbial enumeration technique. I. The persistence of drug-susceptible tubercle bacilli in the tissues despite prolonged antimicrobial therapy. J. Exp. Med. 104 737–762. 10.1084/jem.104.5.737
- Mitchison D. A. (1985). The action of antituberculosis drugs in short-course chemotherapy. Tubercle 66 219–225. 10.1016/0041-3879(85)90040-6
- Mitchison D. A. (1998). Development of rifapentine: the way ahead. Int. J. Tuberc. Lung Dis. 2 612–615.
- Mitchison D. A. (2000). Role of individual drugs in the chemotherapy of tuberculosis. Int. J. Tuberc. Lung Dis. 4 796–806.
- Mitchison D. A. (2005). Shortening the treatment of tuberculosis. Nat. Biotechnol. 23 187–188. 10.1038/nbt0205-187
- Mitchison D. A. (2012). Pharmacokinetic/pharmacodynamic parameters and the choice of high-dosage rifamycins. Int. J. Tuberc. Lung Dis. 16 1186–1189. 10.5588/ijtld.11.0818
- Mitchison D. A., Fourie P. B. (2010). The near future: improving the activity of rifamycins and pyrazinamide. Tuberculosis (Edinb.) 90 177–181. 10.1016/j.tube.2010.03.005
- Mukamolova G. V., Kaprelyants A. S., Young D. I., Young M., Kell D. B. (1998). A bacterial cytokine. Proc. Natl. Acad. Sci. U.S.A. 95 8916–8921. 10.1073/pnas.95.15.8916
- Mukamolova G. V., Turapov O., Malkin J., Woltmann G., Barer M. R. (2010). Resuscitation-promoting factors reveal an occult population of tubercle Bacilli in Sputum. Am. J. Respir. Crit. Care Med. 181 174–180. 10.1164/rccm.200905-0661OC
- Nunn A. J., Phillips P. P., Gillespie S. H. (2008). Design issues in pivotal drug trials for drug sensitive tuberculosis (TB). Tuberculosis (Edinb.) 88(Suppl. 1) S85–S92. 10.1016/S1472-9792(08)70039-8
- Pasipanodya J. G., McIlleron H., Burger A., Wash P. A., Smith P., Gumbo T. (2013). Serum drug concentrations predictive of pulmonary tuberculosis outcomes. J. Infect. Dis. 208 1464–1473. 10.1093/infdis/jit352
- Rosenthal I. M., Tasneen R., Peloquin C. A., Zhang M., Almeida D., Mdluli K. E., et al. (2012). Dose-ranging comparison of rifampin and rifapentine in two pathologically distinct murine models of tuberculosis. Antimicrob. Agents Chemother. 56 4331–4340. 10.1128/AAC.00912-12
- Shleeva M. O., Bagramyan K., Telkov M. V., Mukamolova G. V., Young M., Kell D. B., et al. (2002). Formation and resuscitation of “non-culturable” cells of Rhodococcus rhodochrous and Mycobacterium tuberculosis in prolonged stationary phase. Microbiology (Read. Engl.) 148 1581–1591.
- Sirgel F. A., Fourie P. B., Donald P. R., Padayatchi N., Rustomjee R., Levin J., et al. (2005). The early bactericidal activities of rifampin and rifapentine in pulmonary tuberculosis. Am. J. Respir. Crit. Care Med. 172 128–135. 10.1164/rccm.200411-1557OC
- van Niekerk C., Ginsberg A. (2009). Assessment of global capacity to conduct tuberculosis drug development trials: do we have what it takes? Int. J. Tuberc. Lung Dis. 13 1367–1372.
- Wayne L. G. (1976). Dynamics of submerged growth of Mycobacterium tuberculosis under aerobic and microaerophilic conditions. Am. Rev. Respir. Dis. 114 807–811.
- Wayne L. G. (1977). Synchronized replication of Mycobacterium tuberculosis. Infect. Immun. 17 528–530.
- Wayne L. G. (1994). Dormancy of Mycobacterium tuberculosis and latency of disease. Eur. J. Clin. Microbiol. Infect. Dis. 13 908–914. 10.1007/BF02111491
- World Health Organization [WHO]. (2010). WHO global tuberculosis control report 2010. Summary. Cent. Eur. J. Public Health 18 237.
- Yew W. W., Leung C. C. (2006). Antituberculosis drugs and hepatotoxicity. Respirology 11 699–707. 10.1111/j.1440-1843.2006.00941.x
Source: PubMed