{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T10:50:54Z","timestamp":1792752654455,"version":"4.5.0"},"posted":{"date-parts":[[2024,12,18]]},"group-title":"Microbiology","reference-count":48,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,12,18]],"date-time":"2024-12-18T00:00:00Z","timestamp":1734480000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,12,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Pyrazinamide (PZA) is a critical component of tuberculosis first-line therapy due to its ability to kill both growing and non-replicating drug-tolerant populations of\n                  <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                  within the host. Recent evidence indicates that PZA acts through disruption of coenzyme A synthesis under conditions that promote cellular stress. In contrast to its bactericidal action\n                  <jats:italic>in vivo<\/jats:italic>\n                  , PZA shows weak bacteriostatic activity against\n                  <jats:italic>M. tuberculosis<\/jats:italic>\n                  in axenic culture. While the basis for this striking difference between\n                  <jats:italic>in vivo<\/jats:italic>\n                  and\n                  <jats:italic>in vitro<\/jats:italic>\n                  PZA activity has yet to be resolved, recent studies have highlighted an important role for cell-mediated immunity in PZA efficacy. These observations suggest that host-derived antimicrobial activity may contribute to the bactericidal action of PZA within the host environment. In this study we show that the active form of PZA, pyrazinoic acid (POA), synergizes with the bactericidal activity of host-derived reactive oxygen species (ROS). We determined that POA can promote increased cellular oxidative damage and enhanced killing of\n                  <jats:italic>M. tuberculosis<\/jats:italic>\n                  . Further, we find that the thiol oxidant diamide is also able to potentiate PZA activity, implicating thiol oxidation as a key driver of PZA susceptibility. Using a macrophage infection model, we demonstrate the essentiality of interferon-\u03b3 induced ROS production for PZA mediated clearance of\n                  <jats:italic>M. tuberculosis<\/jats:italic>\n                  . Based on these observations, we propose that the\n                  <jats:italic>in vivo<\/jats:italic>\n                  sterilizing activity of PZA can be mediated through its synergistic interaction with the host oxidative burst leading to collateral disruption of coenzyme A metabolism. These findings will enable discovery efforts to identify novel host- and microbe-directed approaches to bolster PZA efficacy.\n                <\/jats:p>","DOI":"10.1101\/2024.12.17.628853","type":"posted-content","created":{"date-parts":[[2024,12,18]],"date-time":"2024-12-18T08:20:17Z","timestamp":1734510017000},"source":"Crossref","is-referenced-by-count":4,"title":["Oxidative stress drives potent bactericidal activity of pyrazinamide against\n                  <i>Mycobacterium tuberculosis<\/i>"],"prefix":"10.64898","author":[{"given":"Nicholas A.","family":"Dillon","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elise A.","family":"Lamont","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muzafar A.","family":"Rather","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1188-4238","authenticated-orcid":false,"given":"Anthony D.","family":"Baughn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2024122100571749000_2024.12.17.628853v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2006.09.004"},{"key":"2024122100571749000_2024.12.17.628853v1.2","first-page":"145","article-title":"Revisiting Anti-tuberculosis Activity of Pyrazinamide in Mice","volume":"4","year":"2014","journal-title":"Mycobact Dis"},{"key":"2024122100571749000_2024.12.17.628853v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.cotox.2018.03.005"},{"key":"2024122100571749000_2024.12.17.628853v1.4","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00529-10"},{"key":"2024122100571749000_2024.12.17.628853v1.5","doi-asserted-by":"crossref","unstructured":"Berry, S. B. , S. Espich , N. T. T. Thuong , X. Chang , R. Dorajoo , C.-C. Khor , C.-K. Heng , J.-M. Yuan , D. Fox , A. Anaya-Sanchez , L. Tenney , C. J. Chang , D. I. Kotov , R. E. Vance , S. J. Dunstan , K. H. Darwin and S. A. Stanley (2023). \u201cDisruption of Aldehyde Dehydrogenase 2 protects against bacterial infection.\u201d bioRxiv: 2023.2008.2024.554661.","DOI":"10.1101\/2023.08.24.554661"},{"key":"2024122100571749000_2024.12.17.628853v1.6","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M800694200"},{"key":"2024122100571749000_2024.12.17.628853v1.7","unstructured":"British-Medical-Research-Council (1974). \u201cControlled Clinical Trial of Four Short-Course (6-month) Regimens of Chemotherapy for Treatment of Pulmonary Tuberculosis. Third report.\u201d Lancet: 237\u2013240."},{"key":"2024122100571749000_2024.12.17.628853v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2007.04.034"},{"key":"2024122100571749000_2024.12.17.628853v1.9","doi-asserted-by":"publisher","DOI":"10.1084\/jem.175.4.1111"},{"key":"2024122100571749000_2024.12.17.628853v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2017.06.018"},{"key":"2024122100571749000_2024.12.17.628853v1.11","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.04028-14"},{"key":"2024122100571749000_2024.12.17.628853v1.12","doi-asserted-by":"publisher","DOI":"10.4049\/immunohorizons.2200077"},{"key":"2024122100571749000_2024.12.17.628853v1.13","doi-asserted-by":"publisher","DOI":"10.1084\/jem.178.6.2249"},{"key":"2024122100571749000_2024.12.17.628853v1.14","doi-asserted-by":"publisher","DOI":"10.1016\/1074-7613(95)90001-2"},{"key":"2024122100571749000_2024.12.17.628853v1.15","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.7b00079"},{"key":"2024122100571749000_2024.12.17.628853v1.16","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.7b00079"},{"key":"2024122100571749000_2024.12.17.628853v1.17","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-15516-1"},{"key":"2024122100571749000_2024.12.17.628853v1.18","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.7b00017"},{"issue":"3","key":"2024122100571749000_2024.12.17.628853v1.19","first-page":"317","article-title":"Sterilising action of pyrazinamide in models of dormant and rifampicin-tolerant Mycobacterium tuberculosis","volume":"10","year":"2006","journal-title":"Int J Tuberc Lung Dis"},{"key":"2024122100571749000_2024.12.17.628853v1.20","doi-asserted-by":"publisher","DOI":"10.1515\/bmc-2018-0007"},{"key":"2024122100571749000_2024.12.17.628853v1.21","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccr.2012.09.012"},{"issue":"2","key":"2024122100571749000_2024.12.17.628853v1.22","first-page":"e00070","article-title":"The Bewildering Antitubercular Action of Pyrazinamide","volume":"84","year":"2020","journal-title":"Microbiol Mol Biol Rev"},{"key":"2024122100571749000_2024.12.17.628853v1.23","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01370-15"},{"key":"2024122100571749000_2024.12.17.628853v1.24","doi-asserted-by":"publisher","DOI":"10.1242\/dmm.019513"},{"key":"2024122100571749000_2024.12.17.628853v1.25","doi-asserted-by":"publisher","DOI":"10.1097\/INF.0b013e31815b494c"},{"key":"2024122100571749000_2024.12.17.628853v1.26","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.74358"},{"key":"2024122100571749000_2024.12.17.628853v1.27","doi-asserted-by":"crossref","unstructured":"MacMicking, J. D. (2014). \u201cCell-autonomous effector mechanisms against mycobacterium tuberculosis.\u201d Cold Spring Harb Perspect Med 4(10).","DOI":"10.1101\/cshperspect.a018507"},{"key":"2024122100571749000_2024.12.17.628853v1.28","doi-asserted-by":"publisher","DOI":"10.1084\/jem.158.3.670"},{"key":"2024122100571749000_2024.12.17.628853v1.29","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkg301"},{"key":"2024122100571749000_2024.12.17.628853v1.30","doi-asserted-by":"publisher","DOI":"10.1016\/0007-0971(87)90160-4"},{"key":"2024122100571749000_2024.12.17.628853v1.31","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0401657101"},{"key":"2024122100571749000_2024.12.17.628853v1.32","doi-asserted-by":"publisher","DOI":"10.1038\/nm.3937"},{"key":"2024122100571749000_2024.12.17.628853v1.33","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-03452-y"},{"key":"2024122100571749000_2024.12.17.628853v1.34","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.00117-22"},{"key":"2024122100571749000_2024.12.17.628853v1.35","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-24127-3"},{"key":"2024122100571749000_2024.12.17.628853v1.36","first-page":"e58","article-title":"Aspartate decarboxylase (PanD) as a new target of pyrazinamide in Mycobacterium tuberculosis","volume":"3","year":"2014","journal-title":"Emerg Microbes Infect"},{"key":"2024122100571749000_2024.12.17.628853v1.37","doi-asserted-by":"publisher","DOI":"10.1006\/jcat.1993.1196"},{"key":"2024122100571749000_2024.12.17.628853v1.38","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.41.3.636"},{"key":"2024122100571749000_2024.12.17.628853v1.39","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-14238-3"},{"key":"2024122100571749000_2024.12.17.628853v1.40","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.00439-21"},{"key":"2024122100571749000_2024.12.17.628853v1.41","doi-asserted-by":"publisher","DOI":"10.1016\/S2213-2600(19)30366-2"},{"key":"2024122100571749000_2024.12.17.628853v1.42","doi-asserted-by":"publisher","DOI":"10.1016\/j.freeradbiomed.2015.03.008"},{"key":"2024122100571749000_2024.12.17.628853v1.43","doi-asserted-by":"crossref","unstructured":"VanderVen, B. C. , L. Huang , K. H. Rohde and D. G. Russell (2016). \u201cThe Minimal Unit of Infection: Mycobacterium tuberculosis in the Macrophage.\u201d Microbiol Spectr 4(6).","DOI":"10.1128\/microbiolspec.TBTB2-0025-2016"},{"key":"2024122100571749000_2024.12.17.628853v1.44","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2010.07099.x"},{"key":"2024122100571749000_2024.12.17.628853v1.45","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-62703-239-1_18"},{"key":"2024122100571749000_2024.12.17.628853v1.46","doi-asserted-by":"publisher","DOI":"10.1002\/cpcb.60"},{"key":"2024122100571749000_2024.12.17.628853v1.47","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.200501"},{"key":"2024122100571749000_2024.12.17.628853v1.48","first-page":"523","article-title":"Pyrazinamide (Aldinamide) in the treatment of pulmonary tuberculosis","volume":"65","year":"1952","journal-title":"American Review of tuberculosis"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2024.12.17.628853","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T07:42:01Z","timestamp":1768462921000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2024.12.17.628853"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,18]]},"references-count":48,"URL":"https:\/\/doi.org\/10.1101\/2024.12.17.628853","relation":{"has-version":[{"id-type":"doi","id":"10.7554\/eLife.105614.1","asserted-by":"object"}]},"subject":[],"published":{"date-parts":[[2024,12,18]]},"subtype":"preprint"}}