{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T15:02:20Z","timestamp":1786719740645,"version":"build-2736575974"},"reference-count":45,"publisher":"American Chemical Society (ACS)","issue":"8","license":[{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100000032","name":"Government of Canada","doi-asserted-by":"publisher","award":["152931"],"award-info":[{"award-number":["152931"]}],"id":[{"id":"10.13039\/501100000032","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000082","name":"Cystic Fibrosis Canada","doi-asserted-by":"publisher","award":["AWD-021084"],"award-info":[{"award-number":["AWD-021084"]}],"id":[{"id":"10.13039\/501100000082","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005247","name":"University of British Columbia","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005247","id-type":"DOI","asserted-by":"publisher"}]},{"name":"TB Veterans Charitable Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,14]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Treatments for tuberculosis disease (TB) are failing on a global scale, necessitating new drug discovery efforts to identify effective therapies. Mycobacterium tuberculosis (Mtb), the obligate human pathogen and causative agent of TB, resides within alveolar macrophages and combats acidic stress by inhibiting phagosome maturation. Understanding the role of acidification in Mtb pathogenesis can aid the development of improved TB therapies. Aligning with the hypothesis that acidification affects Mtb intracellular survival, we developed a phenotypic assay to identify compounds that enhance acidification of Mtb in infected macrophages. To evaluate the assay suitability for screening purposes, we screened two compound libraries and found that early acidification does not necessarily correlate with inhibition of Mtb growth over time. Further, we found that compounds interfering with bacterial protein and DNA synthesis induced early phagosome acidification. However, other compounds, including cell wall synthesis inhibitors, did not consistently correlate with acidification or growth inhibition. Using early acidification could therefore provide a potential avenue to inform antimicrobial kinetics. By assessing Mtb\u2019s response to phagosome acidification, this work highlights the complexity of host\u2013pathogen interactions in early infection and provides a valuable tool for investigating Mtb survival mechanisms and accelerating drug discovery efforts.<\/jats:p>","DOI":"10.1021\/acsinfecdis.6c00093","type":"journal-article","created":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T16:46:58Z","timestamp":1782924418000},"page":"2650-2662","source":"Crossref","is-referenced-by-count":0,"title":["Comparative\nPhenotypic Screening Identifies Protein\nSynthesis Inhibitors as Compounds That Enhance Early Acidification\nof\n                    <i>Mycobacterium tuberculosis<\/i>\n                    in Macrophages"],"prefix":"10.1021","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0049-5408","authenticated-orcid":true,"given":"Leah","family":"Rankine-Wilson","sequence":"first","affiliation":[{"name":"University of British Columbia , , ,","place":["Vancouver, Canada, V6T 1Z4"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9547-0237","authenticated-orcid":true,"given":"Yossef","family":"Av-Gay","sequence":"additional","affiliation":[{"name":"University of British Columbia , , ,","place":["Vancouver, Canada, V6T 1Z4"]},{"name":"University of British Columbia , , ,","place":["Vancouver, Canada, V6T 1Z3"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,1]]},"reference":[{"issue":"48","key":"2026081410070832200_cit1","doi-asserted-by":"publisher","first-page":"19371","DOI":"10.1073\/pnas.1109201108","article-title":"Mycobacterium\nTuberculosis Protein Tyrosine Phosphatase (PtpA) Excludes Host Vacuolar-H+-ATPase\nto Inhibit Phagosome Acidification","volume":"108","author":"Wong","year":"2011","journal-title":"Proc. Natl.\nAcad. Sci. U. S. A."},{"issue":"7","key":"2026081410070832200_cit2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0070514","article-title":"Secreted\nAcid Phosphatase (SapM)\nof Mycobacterium Tuberculosis Is Indispensable for Arresting Phagosomal\nMaturation and Growth of the Pathogen in Guinea Pig Tissues","volume":"8","author":"Puri","year":"2013","journal-title":"PLoS One"},{"issue":"1","key":"2026081410070832200_cit3","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1046\/j.1365-2958.2001.02500.x","article-title":"An Essential Role for phoP in Mycobacterium\nTuberculosis\nVirulence","volume":"41","author":"Pe\u0301rez","year":"2001","journal-title":"Mol. Microbiol."},{"issue":"8","key":"2026081410070832200_cit4","doi-asserted-by":"publisher","first-page":"849","DOI":"10.1038\/nm.1795","article-title":"A Membrane\nProtein Preserves Intrabacterial pH in Intraphagosomal Mycobacterium\nTuberculosis","volume":"14","author":"Vandal","year":"2008","journal-title":"Nat. Med."},{"issue":"7","key":"2026081410070832200_cit5","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0068942","article-title":"Whole Cell\nScreen for Inhibitors of pH Homeostasis in Mycobacterium Tuberculosis","volume":"8","author":"Darby","year":"2013","journal-title":"PLoS One"},{"issue":"9","key":"2026081410070832200_cit6","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1001100","article-title":"High Content\nPhenotypic Cell-Based Visual Screen Identifies Mycobacterium Tuberculosis\nAcyltrehalose-Containing Glycolipids Involved in Phagosome Remodeling","volume":"6","author":"Brodin","year":"2010","journal-title":"PLoS Pathog."},{"issue":"37","key":"2026081410070832200_cit7","doi-asserted-by":"publisher","first-page":"13642","DOI":"10.1073\/pnas.0401657101","article-title":"Isolation of Mycobacterium\nTuberculosis Mutants Defective\nin the Arrest of Phagosome Maturation","volume":"101","author":"Pethe","year":"2004","journal-title":"Proc.\nNatl. Acad. Sci. U. S. A."},{"issue":"2","key":"2026081410070832200_cit8","doi-asserted-by":"publisher","first-page":"272","DOI":"10.1021\/acsinfecdis.8b00256","article-title":"Identification\nof Compounds with pH-Dependent Bactericidal Activity against Mycobacterium\nTuberculosis","volume":"5","author":"Early","year":"2019","journal-title":"ACS Infect. Dis."},{"issue":"3","key":"2026081410070832200_cit9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0010033","article-title":"Mycobacterial\nMutants with Defective Control of Phagosomal Acidification","volume":"1","author":"Stewart","year":"2005","journal-title":"PLOS Pathog."},{"issue":"2","key":"2026081410070832200_cit10","doi-asserted-by":"publisher","first-page":"250","DOI":"10.1016\/j.chom.2016.07.007","article-title":"Mycobacterial Acid Tolerance Enables\nPhagolysosomal\nSurvival and Establishment of Tuberculous Infection In Vivo","volume":"20","author":"Levitte","year":"2016","journal-title":"Cell Host Microbe"},{"issue":"6","key":"2026081410070832200_cit11","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00226-17","article-title":"Lung Tissue\nConcentrations of Pyrazinamide among Patients with Drug-Resistant\nPulmonary Tuberculosis","volume":"61","author":"Kempker","year":"2017","journal-title":"Antimicrob. Agents Chemother."},{"issue":"32","key":"2026081410070832200_cit12","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2024571118","article-title":"Growth of Mycobacterium Tuberculosis at Acidic pH Depends on Lipid\nAssimilation and Is Accompanied by Reduced GAPDH Activity","volume":"118","author":"Gouzy","year":"2021","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"issue":"1","key":"2026081410070832200_cit13","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-22343-4","article-title":"Genetic\nand Metabolic Regulation of Mycobacterium Tuberculosis\nAcid Growth Arrest","volume":"8","author":"Baker","year":"2018","journal-title":"Sci. Rep."},{"issue":"5","key":"2026081410070832200_cit14","doi-asserted-by":"publisher","first-page":"3031","DOI":"10.1128\/IAI.72.5.3031-3037.2004","article-title":"Protection\nElicited by a Double Leucine\nand Pantothenate Auxotroph of Mycobacterium Tuberculosis in Guinea\nPigs","volume":"72","author":"Sampson","year":"2004","journal-title":"Infect. Immun."},{"key":"2026081410070832200_cit15","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.01922","article-title":"Comprehensive Characterization\nof the Attenuated Double\nAuxotroph Mycobacterium tuberculosis \u0394leuD\u0394panCD as an Alternative to H37Rv","volume":"10","author":"Mouton","year":"2019","journal-title":"Front. Microbiol."},{"issue":"5","key":"2026081410070832200_cit16","doi-asserted-by":"publisher","first-page":"352","DOI":"10.1016\/j.chom.2007.09.006","article-title":"Mycobacterium Tuberculosis\nInvasion\nof Macrophages: Linking Bacterial Gene Expression to Environmental\nCues","volume":"2","author":"Rohde","year":"2007","journal-title":"Cell Host Microbe"},{"issue":"9","key":"2026081410070832200_cit17","doi-asserted-by":"publisher","first-page":"1000","DOI":"10.1177\/2472555220942764","article-title":"Z\u2019 Does Not\nNeed to Be &gt; 0.5","volume":"25","author":"Bar","year":"2020","journal-title":"SLAS Discovery"},{"key":"2026081410070832200_cit18","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1007\/978-1-0716-1971-1_7","article-title":"Mycobacterium Tuberculosis\nInfection of THP-1 Cells: A Model for High Content Analysis of Intracellular\nGrowth and Drug Susceptibility","volume":"2427","author":"Rankine-Wilson","year":"2022","journal-title":"Methods Mol.\nBiol. Clifton NJ."},{"issue":"23","key":"2026081410070832200_cit19","doi-asserted-by":"publisher","first-page":"3815","DOI":"10.1093\/bioinformatics\/btv455","article-title":"Impact of Normalization Methods on\nHigh-Throughput\nScreening Data with High Hit Rates and Drug Testing with Dose\u2013Response\nData","volume":"31","author":"Mpindi","year":"2015","journal-title":"Bioinformatics"},{"issue":"1","key":"2026081410070832200_cit20","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1074\/jbc.M111.302588","article-title":"The Mycobacterial Transcriptional\nRegulator whiB7 Gene Links Redox Homeostasis and Intrinsic Antibiotic\nResistance","volume":"287","author":"Burian","year":"2012","journal-title":"J. Biol. Chem."},{"issue":"8","key":"2026081410070832200_cit21","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00283-18","article-title":"Rifabutin\nActs in Synergy and Is\nBactericidal with Frontline Mycobacterium Abscessus Antibiotics Clarithromycin\nand Tigecycline, Suggesting a Potent Treatment Combination","volume":"62","author":"Pryjma","year":"2018","journal-title":"Antimicrob. Agents Chemother."},{"key":"2026081410070832200_cit22","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.678354","article-title":"Stabilization of Hypoxia-Inducible Factor Promotes\nAntimicrobial Activity of Human Macrophages Against Mycobacterium\nTuberculosis","volume":"12","author":"Zenk","year":"2021","journal-title":"Front. Immunol."},{"issue":"12","key":"2026081410070832200_cit23","doi-asserted-by":"publisher","first-page":"3593","DOI":"10.1158\/0008-5472.CAN-16-0061","article-title":"The Small Molecule IMR-1 Inhibits the Notch Transcriptional\nActivation Complex to Suppress Tumorigenesis","volume":"76","author":"Astudillo","year":"2016","journal-title":"Cancer Res."},{"issue":"1","key":"2026081410070832200_cit24","doi-asserted-by":"publisher","first-page":"68","DOI":"10.1016\/S0165-5728(99)00134-4","article-title":"Beta2-Adrenergic\nReceptor Stimulation\nInhibits Nitric Oxide Generation by Mycobacterium Avium Infected Macrophages","volume":"101","author":"Boomershine","year":"1999","journal-title":"J. Neuroimmunol."},{"issue":"12","key":"2026081410070832200_cit25","doi-asserted-by":"publisher","DOI":"10.15252\/embj.201798694","article-title":"LRRK2 Is a Negative Regulator of\nMycobacterium Tuberculosis\nPhagosome Maturation in Macrophages","volume":"37","author":"Ha\u0308rtlova","year":"2018","journal-title":"EMBO J."},{"issue":"11","key":"2026081410070832200_cit26","doi-asserted-by":"publisher","first-page":"942","DOI":"10.1016\/j.tim.2019.06.005","article-title":"Acid Fasting: Modulation of Mycobacterium\nTuberculosis Metabolism at Acidic pH","volume":"27","author":"Baker","year":"2019","journal-title":"Trends\nMicrobiol."},{"issue":"1","key":"2026081410070832200_cit27","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1111\/j.1600-065X.2007.00547.x","article-title":"Mycobacterium\nTuberculosis\nand the Environment within the Phagosome","volume":"219","author":"Rohde","year":"2007","journal-title":"Immunol.\nRev."},{"issue":"4","key":"2026081410070832200_cit28","doi-asserted-by":"publisher","DOI":"10.1128\/mSystems.00057-17","article-title":"Integration of Metabolomics\nand Transcriptomics Reveals a Complex Diet of Mycobacterium Tuberculosis\nduring Early Macrophage Infection","volume":"2","author":"Zimmermann","year":"2017","journal-title":"mSystems"},{"key":"2026081410070832200_cit29","unstructured":"Canada, P. H. A.\n          \n          Chapter\n5: Canadian Tuberculosis Standards 7th Edition: 2014 \u2013 Treatment\nof Tuberculosis disease, https:\/\/www.canada.ca\/en\/public-health\/services\/infectious-diseases\/canadian-tuberculosis-standards-7th-edition\/edition-17.html (accessed Oct 03, 2024)."},{"key":"2026081410070832200_cit30","doi-asserted-by":"publisher","first-page":"1634","DOI":"10.1002\/1873-3468.70023","article-title":"Unraveling Mycobacterium\nTuberculosis Acid Resistance and pH Homeostasis Mechanisms","volume":"599","author":"Laudouze","year":"2025","journal-title":"FEBS Lett."},{"issue":"32","key":"2026081410070832200_cit31","doi-asserted-by":"publisher","first-page":"9975","DOI":"10.1021\/bi0005815","article-title":"Catalase-Peroxidase (Mycobacterium\nTuberculosis KatG) Catalysis and Isoniazid Activation","volume":"39","author":"Chouchane","year":"2000","journal-title":"Biochemistry"},{"issue":"18","key":"2026081410070832200_cit32","doi-asserted-by":"publisher","first-page":"3450","DOI":"10.1016\/j.jmb.2019.02.016","article-title":"The Isoniazid\nParadigm of Killing, Resistance, and\nPersistence in Mycobacterium Tuberculosis","volume":"431","author":"Vilche\u0300ze","year":"2019","journal-title":"J.\nMol. Biol."},{"issue":"2","key":"2026081410070832200_cit33","doi-asserted-by":"publisher","first-page":"670","DOI":"10.1128\/AAC.47.2.670-675.2003","article-title":"Isoniazid\nActivation Defects in Recombinant\nMycobacterium Tuberculosis Catalase-Peroxidase (KatG) Mutants Evident\nin InhA Inhibitor Production","volume":"47","author":"Wei","year":"2003","journal-title":"Antimicrob. Agents\nChemother."},{"key":"2026081410070832200_cit34","doi-asserted-by":"publisher","first-page":"80","DOI":"10.1016\/j.tube.2017.04.010","article-title":"The\nRole of the Time-Kill Kinetics Assay as Part of a Preclinical Modeling\nFramework for Assessing the Activity of Anti-Tuberculosis Drugs","volume":"105","author":"Bax","year":"2017","journal-title":"Tuberculosis"},{"issue":"12","key":"2026081410070832200_cit35","doi-asserted-by":"publisher","first-page":"2582","DOI":"10.1093\/jac\/dkq374","article-title":"Time\u2013Kill Kinetics of Anti-Tuberculosis\nDrugs, and Emergence of Resistance, in Relation to Metabolic Activity\nof Mycobacterium Tuberculosis","volume":"65","author":"de Steenwinkel","year":"2010","journal-title":"J. Antimicrob.\nChemother."},{"issue":"7","key":"2026081410070832200_cit36","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00280-20","article-title":"Advanced Quantification Methods To Improve the 18b\nDormancy Model for Assessing the Activity of Tuberculosis Drugs In\nVitro","volume":"64","author":"Pieterman","year":"2020","journal-title":"Antimicrob. Agents Chemother."},{"key":"2026081410070832200_cit37","doi-asserted-by":"publisher","DOI":"10.1128\/aac.00478-25","article-title":"Vacuolar-ATPase Inhibitors\nAre Antimicrobial Agents\nActive against Intracellular Mycobacteria","volume":"69","author":"Rankine-Wilson","year":"2025","journal-title":"Antimicrob.\nAgents Chemother."},{"issue":"5","key":"2026081410070832200_cit38","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1007\/BF00295501","article-title":"Erythromycin Inhibits the Assembly of the Large Ribosomal\nSubunit in Growing Escherichia Coli Cells","volume":"30","author":"Chittum","year":"1995","journal-title":"Curr. Microbiol."},{"issue":"3","key":"2026081410070832200_cit39","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1016\/S0025-6196(12)60219-5","article-title":"Erythromycin: A\nMicrobial and Clinical Perspective\nAfter 30 Years of Clinical Use (First of Two Parts)*","volume":"60","author":"Washington","year":"1985","journal-title":"Mayo Clin. Proc."},{"issue":"5","key":"2026081410070832200_cit40","doi-asserted-by":"publisher","first-page":"1005","DOI":"10.1016\/S0022-2836(03)00662-4","article-title":"The Mechanism\nof Action of Macrolides,\nLincosamides and Streptogramin B Reveals the Nascent Peptide Exit\nPath in the Ribosome","volume":"330","author":"Tenson","year":"2003","journal-title":"J. Mol. Biol."},{"issue":"Suppl B","key":"2026081410070832200_cit41","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1093\/jac\/20.suppl_b.47","article-title":"Cellular Uptake and Subcellular Distribution\nof Roxithromycin and Erythromycin in Phagocytic Cells","volume":"20","author":"Carlier","year":"1987","journal-title":"J. Antimicrob. Chemother."},{"issue":"2","key":"2026081410070832200_cit42","doi-asserted-by":"publisher","first-page":"100","DOI":"10.1007\/BF01964420","article-title":"Intracellular\nDistribution and Activity of Antibiotics","volume":"10","author":"Tulkens","year":"1991","journal-title":"Eur.\nJ. Clin. Microbiol. Infect. Dis."},{"issue":"2","key":"2026081410070832200_cit43","doi-asserted-by":"publisher","first-page":"444","DOI":"10.1183\/09031936.00147014","article-title":"Evaluation\nof Macrolides for Possible Use against Multidrug-Resistant Mycobacterium\nTuberculosis","volume":"46","author":"van der Paardt","year":"2015","journal-title":"Eur. Respir. J."},{"key":"2026081410070832200_cit44","doi-asserted-by":"publisher","DOI":"10.1016\/j.pupt.2021.102095","article-title":"The Immunomodulatory\nEffects of Macrolide Antibiotics\nin Respiratory Disease","volume":"71","author":"Pollock","year":"2021","journal-title":"Pulm. Pharmacol. Ther."},{"issue":"3","key":"2026081410070832200_cit45","doi-asserted-by":"publisher","first-page":"590","DOI":"10.1128\/CMR.00078-09","article-title":"Mechanisms of Action\nand Clinical Application of Macrolides\nas Immunomodulatory Medications","volume":"23","author":"Kanoh","year":"2010","journal-title":"Clin. Microbiol.\nRev."}],"container-title":["ACS Infectious Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsinfecdis.6c00093","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/12\/8\/2650\/65313445\/acsinfecdis.6c00093.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article-pdf\/12\/8\/2650\/65313445\/acsinfecdis.6c00093.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T14:17:52Z","timestamp":1786717072000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/aidcbc\/article\/12\/8\/2650\/5172396\/Comparative-Phenotypic-Screening-Identifies"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,1]]},"references-count":45,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,7,1]]},"published-print":{"date-parts":[[2026,8,14]]}},"URL":"https:\/\/doi.org\/10.1021\/acsinfecdis.6c00093","relation":{},"ISSN":["2373-8227"],"issn-type":[{"value":"2373-8227","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,1]]}}}