Reproducibility of the Ribosomal RNA Synthesis Ratio in Sputum and Association with Markers of Mycobacterium tuberculosis Burden
ABSTRACT There is a critical need for improved pharmacodynamic markers for use in human tuberculosis (TB) drug trials. Pharmacodynamic monitoring in TB has conventionally used culture or molecular methods to enumerate the burden of Mycobacterium tuberculosis organisms in sputum. A recently proposed...
| Main Authors: | , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2021-10-01
|
| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/Spectrum.00481-21 |
| _version_ | 1798034101177942016 |
|---|---|
| author | Emmanuel Musisi Christian Dide-Agossou Reem Al Mubarak Karen Rossmassler Abdul Wahab Ssesolo Sylvia Kaswabuli Patrick Byanyima Ingvar Sanyu Josephine Zawedde William Worodria Martin I. Voskuil Rada M. Savic Payam Nahid J. Lucian Davis Laurence Huang Camille M. Moore Nicholas D. Walter |
| author_facet | Emmanuel Musisi Christian Dide-Agossou Reem Al Mubarak Karen Rossmassler Abdul Wahab Ssesolo Sylvia Kaswabuli Patrick Byanyima Ingvar Sanyu Josephine Zawedde William Worodria Martin I. Voskuil Rada M. Savic Payam Nahid J. Lucian Davis Laurence Huang Camille M. Moore Nicholas D. Walter |
| author_sort | Emmanuel Musisi |
| collection | DOAJ |
| description | ABSTRACT There is a critical need for improved pharmacodynamic markers for use in human tuberculosis (TB) drug trials. Pharmacodynamic monitoring in TB has conventionally used culture or molecular methods to enumerate the burden of Mycobacterium tuberculosis organisms in sputum. A recently proposed assay called the rRNA synthesis (RS) ratio measures a fundamentally novel property, how drugs impact ongoing bacterial rRNA synthesis. Here, we evaluated RS ratio as a potential pharmacodynamic monitoring tool by testing pretreatment sputa from 38 Ugandan adults with drug-susceptible pulmonary TB. We quantified the RS ratio in paired pretreatment sputa and evaluated the relationship between the RS ratio and microbiologic and molecular markers of M. tuberculosis burden. We found that the RS ratio was highly repeatable and reproducible in sputum samples. The RS ratio was independent of M. tuberculosis burden, confirming that it measures a distinct new property. In contrast, markers of M. tuberculosis burden were strongly associated with each other. These results indicate that the RS ratio is repeatable and reproducible and provides a distinct type of information from markers of M. tuberculosis burden. IMPORTANCE This study takes a major next step toward practical application of a novel pharmacodynamic marker that we believe will have transformative implications for tuberculosis. This article follows our recent report in Nature Communications that an assay called the rRNA synthesis (RS) ratio indicates the treatment-shortening of drugs and regimens. Distinct from traditional measures of bacterial burden, the RS ratio measures a fundamentally novel property, how drugs impact ongoing bacterial rRNA synthesis. |
| first_indexed | 2024-04-11T20:39:42Z |
| format | Article |
| id | doaj.art-3b3552dabcc340459be5e080e7f479c1 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-11T20:39:42Z |
| publishDate | 2021-10-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-3b3552dabcc340459be5e080e7f479c12022-12-22T04:04:16ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972021-10-019210.1128/Spectrum.00481-21Reproducibility of the Ribosomal RNA Synthesis Ratio in Sputum and Association with Markers of Mycobacterium tuberculosis BurdenEmmanuel Musisi0Christian Dide-Agossou1Reem Al Mubarak2Karen Rossmassler3Abdul Wahab Ssesolo4Sylvia Kaswabuli5Patrick Byanyima6Ingvar Sanyu7Josephine Zawedde8William Worodria9Martin I. Voskuil10Rada M. Savic11Payam Nahid12J. Lucian Davis13Laurence Huang14Camille M. Moore15Nicholas D. Walter16Infectious Disease Research Collaboration, Kampala, UgandaDepartment of Epidemiology, Colorado School of Public Health, Aurora, Colorado, USARocky Mountain Regional VA Medical Center, Aurora, Colorado, USARocky Mountain Regional VA Medical Center, Aurora, Colorado, USAInfectious Disease Research Collaboration, Kampala, UgandaInfectious Disease Research Collaboration, Kampala, UgandaInfectious Disease Research Collaboration, Kampala, UgandaInfectious Disease Research Collaboration, Kampala, UgandaInfectious Disease Research Collaboration, Kampala, UgandaInfectious Disease Research Collaboration, Kampala, UgandaDepartment of Immunology and Microbiology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USAConsortium for Applied Microbial Metrics, Aurora, Colorado, USAConsortium for Applied Microbial Metrics, Aurora, Colorado, USADepartment of Epidemiology of Microbial Diseases, Yale School of Public Health, New Haven, Connecticut, USADivision of Pulmonary and Critical Care Medicine, University of California San Francisco, San Francisco, California, USADivision of Biostatistics and Bioinformatics, National Jewish Health, Denver, Colorado, USARocky Mountain Regional VA Medical Center, Aurora, Colorado, USAABSTRACT There is a critical need for improved pharmacodynamic markers for use in human tuberculosis (TB) drug trials. Pharmacodynamic monitoring in TB has conventionally used culture or molecular methods to enumerate the burden of Mycobacterium tuberculosis organisms in sputum. A recently proposed assay called the rRNA synthesis (RS) ratio measures a fundamentally novel property, how drugs impact ongoing bacterial rRNA synthesis. Here, we evaluated RS ratio as a potential pharmacodynamic monitoring tool by testing pretreatment sputa from 38 Ugandan adults with drug-susceptible pulmonary TB. We quantified the RS ratio in paired pretreatment sputa and evaluated the relationship between the RS ratio and microbiologic and molecular markers of M. tuberculosis burden. We found that the RS ratio was highly repeatable and reproducible in sputum samples. The RS ratio was independent of M. tuberculosis burden, confirming that it measures a distinct new property. In contrast, markers of M. tuberculosis burden were strongly associated with each other. These results indicate that the RS ratio is repeatable and reproducible and provides a distinct type of information from markers of M. tuberculosis burden. IMPORTANCE This study takes a major next step toward practical application of a novel pharmacodynamic marker that we believe will have transformative implications for tuberculosis. This article follows our recent report in Nature Communications that an assay called the rRNA synthesis (RS) ratio indicates the treatment-shortening of drugs and regimens. Distinct from traditional measures of bacterial burden, the RS ratio measures a fundamentally novel property, how drugs impact ongoing bacterial rRNA synthesis.https://journals.asm.org/doi/10.1128/Spectrum.00481-21humanMycobacterium tuberculosissputumassay developmentpharmacodynamics |
| spellingShingle | Emmanuel Musisi Christian Dide-Agossou Reem Al Mubarak Karen Rossmassler Abdul Wahab Ssesolo Sylvia Kaswabuli Patrick Byanyima Ingvar Sanyu Josephine Zawedde William Worodria Martin I. Voskuil Rada M. Savic Payam Nahid J. Lucian Davis Laurence Huang Camille M. Moore Nicholas D. Walter Reproducibility of the Ribosomal RNA Synthesis Ratio in Sputum and Association with Markers of Mycobacterium tuberculosis Burden Microbiology Spectrum human Mycobacterium tuberculosis sputum assay development pharmacodynamics |
| title | Reproducibility of the Ribosomal RNA Synthesis Ratio in Sputum and Association with Markers of Mycobacterium tuberculosis Burden |
| title_full | Reproducibility of the Ribosomal RNA Synthesis Ratio in Sputum and Association with Markers of Mycobacterium tuberculosis Burden |
| title_fullStr | Reproducibility of the Ribosomal RNA Synthesis Ratio in Sputum and Association with Markers of Mycobacterium tuberculosis Burden |
| title_full_unstemmed | Reproducibility of the Ribosomal RNA Synthesis Ratio in Sputum and Association with Markers of Mycobacterium tuberculosis Burden |
| title_short | Reproducibility of the Ribosomal RNA Synthesis Ratio in Sputum and Association with Markers of Mycobacterium tuberculosis Burden |
| title_sort | reproducibility of the ribosomal rna synthesis ratio in sputum and association with markers of mycobacterium tuberculosis burden |
| topic | human Mycobacterium tuberculosis sputum assay development pharmacodynamics |
| url | https://journals.asm.org/doi/10.1128/Spectrum.00481-21 |
| work_keys_str_mv | AT emmanuelmusisi reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT christiandideagossou reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT reemalmubarak reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT karenrossmassler reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT abdulwahabssesolo reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT sylviakaswabuli reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT patrickbyanyima reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT ingvarsanyu reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT josephinezawedde reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT williamworodria reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT martinivoskuil reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT radamsavic reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT payamnahid reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT jluciandavis reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT laurencehuang reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT camillemmoore reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden AT nicholasdwalter reproducibilityoftheribosomalrnasynthesisratioinsputumandassociationwithmarkersofmycobacteriumtuberculosisburden |