This study aims to: (1) develop new markers for response to tuberculosis (TB) treatment through expression profiling or analysis of patterns of RNA expression in human blood as quantified by microarray technology and (2) determine whether small regulatory RNA (sRNA) secreted by M tuberculosis act to suppress expression of host-protective genes in macrophages through RNA interference (RNAi). New markers for response to TB treatment. Specific disease states may produce signature RNA patterns not seen in other conditions and the pathogen-specific component of human expression may distinguish between infections. Identifying expression that is specific to TB requires a clinically-relevant comparison group. To avoid over-estimating test accuracy, biomarkers should be developed in the patients likely to be tested in practice with an """"""""intention-to-test"""""""" approach. The first step of this project will identify and validate a TB expression panel by comparison with patients with community-acquired pneumonia (CAP) in cohorts of subjects in the U.S. and Uganda. The second step will test whether expression of the TB panel in blood resolves with successful treatment of active TB. We hypothesize that expression of genes in the TB panel will change to resemble expression in controls without pneumonia, reflecting resolving immune response as bacillary burden decreases. Finally, it is unclear whether blood is a surrogate for understanding TB-specific responses in the lung. Although blood expression may identify effective biomarkers even without recapitulating lung expression, interpretation of the pathobiological significance of blood expression demands understanding the relationship between blood and lung expression. To determine the extent to which TB-specific expression in blood reflects inflammatory pathways in the lung this study will compare expression in simultaneously collected blood and BAL while adjusting for the different cell types present in each specimen type. sRNA secreted by Mtb may subvert host-protective macrophage responses via RNAi. RNAi, the biological process in which endogenous non-coding sRNA molecules bind mRNA and inhibit gene expression, is a fundamental regulatory mechanism in both Mtb and humans. sRNA secreted by Mtb was recently shown to increase bacterial viability in macrophages. We hypothesize that the mechanism for this increased viability is Mtb-derived sRNA silencing of expression of important human host-protective genes. Specifically, through RNA sequencing we have identified Mtb-derived sRNA which are predicted to bind the 3'untranslated region of key host-protective human mRNA transcripts for IkB?, TRAF-3 and TRAF-5. We propose to validate this novel form of cross-species RNAi. If confirmed, this would represent a novel form of host-pathogen interaction that could offer new targets for anti-TB therapy.

Public Health Relevance

Tuberculosis (TB) is an important health concern for current service members and veterans because service in conflict zones can cause TB exposures. For example, rates of TB in Iraq and Afghanistan are 16- and 45-fold higher than in the U.S., respectively, and management of TB risk is a major challenge for military public health. The global emergence of drug-resistant forms of TB makes this risk even greater. Military occupational exposures are a CSR&D priority research area. This proposal aims to develop new biomarkers for TB which would enhance veteran health for several reasons. New TB drugs are urgently needed to shorten treatment and treat drug resistance but clinical trials are held back by lack of an early marker for treatment failure. A new marker would change the clinical trial landscape and could also be used to monitor individual patients. An additional aim is validating a new form of host-pathogen interaction through RNA interference. If confirmed, this mechanism could provide new targets for therapy.

National Institute of Health (NIH)
Veterans Affairs (VA)
Veterans Administration (IK2)
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Infectious Diseases B (INFB)
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VA Eastern Colorado Health Care System
United States
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Walter, Nicholas D; Moore, Camille M; Kayigire, Xavier A et al. (2018) Does discovery of differentially culturable M tuberculosis really demand a new treatment paradigm? Longitudinal analysis of DNA clearance from sputum. BMC Infect Dis 18:293
Ofori-Anyinam, Boatema; Dolganov, Gregory; Van, Tran et al. (2017) Significant under expression of the DosR regulon in M. tuberculosis complex lineage 6 in sputum. Tuberculosis (Edinb) 104:58-64
Zhao, Xueyan; Epperson, L Elaine; Hasan, Nabeeh A et al. (2017) Complete Genome Sequence of Mycobacterium avium subsp. hominissuis Strain H87 Isolated from an Indoor Water Sample. Genome Announc 5:
Walter, Nicholas D; Reves, Randall; Davis, J Lucian (2016) Blood transcriptional signatures for tuberculosis diagnosis: a glass half-empty perspective. Lancet Respir Med 4:e28
Garcia, Benjamin J; Loxton, Andre G; Dolganov, Gregory M et al. (2016) Sputum is a surrogate for bronchoalveolar lavage for monitoring Mycobacterium tuberculosis transcriptional profiles in TB patients. Tuberculosis (Edinb) 100:89-94
Walter, Nicholas D; de Jong, Bouke C; Garcia, Benjamin J et al. (2016) Adaptation of Mycobacterium tuberculosis to Impaired Host Immunity in HIV-Infected Patients. J Infect Dis 214:1205-11
Katagira, Winceslaus; Walter, Nicholas D; Den Boon, Saskia et al. (2016) Empiric TB Treatment of Severely Ill Patients With HIV and Presumed Pulmonary TB Improves Survival. J Acquir Immune Defic Syndr 72:297-303
Philley, Julie V; DeGroote, Mary Ann; Honda, Jennifer R et al. (2016) Treatment of Non-Tuberculous Mycobacterial Lung Disease. Curr Treat Options Infect Dis 8:275-296
Walter, Nicholas D; Miller, Mikaela A; Vasquez, Joshua et al. (2016) Blood Transcriptional Biomarkers for Active Tuberculosis among Patients in the United States: a Case-Control Study with Systematic Cross-Classifier Evaluation. J Clin Microbiol 54:274-82
Walter, Nicholas D; Dolganov, Gregory M; Garcia, Benjamin J et al. (2015) Transcriptional Adaptation of Drug-tolerant Mycobacterium tuberculosis During Treatment of Human Tuberculosis. J Infect Dis 212:990-8

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