Approximately one-third of the world's population is infected with Mycobacterium tuberculosis (M.tb), and the World Health Organization estimates that in 2007 there were 9.27 million new cases and more than 1.7 million people died of tuberculosis. M.tb is able to establish this enormous worldwide burden of disease by subverting innate and adaptive defenses of the host. One way in which it does this is to convert the normally hostile environment of a macrophage into a niche in which it can effectively replicate. Normally during phagosome maturation, the bacterial vacuole is transformed from a comparatively inert compartment to a phagolysosome, an effective microbicidal and degradative compartment. However, a variety of mycobacterial species prevent the normal maturation of the phagosome, residing in a replicative niche that resembles an early endosome, although exactly how they do this is not clear. We hypothesize that to promote its intracellular survival M.tb secrete EsxH in order to inhibit the endosomal sorting complex required for transport (ESCRT), cellular machinery of the macrophage involved in protein trafficking. We found that the ESCRT machinery represents a major vulnerability of the cell, as it is required to control growth of non-pathogens, like Mycobacterium smegmatis, as well as of M.tb. Moreover, we identified a novel host-pathogen interaction between the M.tb protein, EsxH, and the host protein hepatocyte growth factor-regulated tyrosine kinase substrate (Hgs/Hrs), which is a component of the ESCRT machinery. In this proposal, we seek to extend our preliminary studies in order to characterize the mechanism by which ESCRT protects against bacterial infection. In addition, we will investigate the interaction between EsxH and Hgs and evaluate its importance to the outcome of infection. These studies will provide important insight into how M.tb subverts the normal anti-microbial capacity of macrophages. If we understood how M.tb does this, we might be able to improve the mycobacterial killing capacity of the infected macrophage, enabling development of novel therapeutics that have the potential to significantly shorten therapy and change the face of the global epidemic.

Public Health Relevance

The success of Mycobacterium tuberculosis (M.tb) as a pathogen is due to its ability to survive within macrophages, cells that normally eradicate bacteria. How it does this is poorly understood. We found that ESCRT, a host cell protein complex, protects macrophages against non-pathogenic mycobacteria, and we intend to characterize its role in M.tb infection. This will advance our understanding of how M.tb survives in macrophages, which might enable the creation of much needed, new therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI087682-05
Application #
9125720
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Eichelberg, Katrin
Project Start
2015-07-01
Project End
2017-06-30
Budget Start
2016-07-01
Budget End
2017-06-30
Support Year
5
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Washington University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Upadhyay, S; Mittal, E; Philips, J A (2018) Tuberculosis and the art of macrophage manipulation. Pathog Dis 76:
Mittal, Ekansh; Skowyra, Michael L; Uwase, Grace et al. (2018) Mycobacterium tuberculosis Type VII Secretion System Effectors Differentially Impact the ESCRT Endomembrane Damage Response. MBio 9:
Portal-Celhay, Cynthia; Tufariello, JoAnn M; Srivastava, Smita et al. (2016) Mycobacterium tuberculosis EsxH inhibits ESCRT-dependent CD4+ T-cell activation. Nat Microbiol 2:16232
Ouimet, Mireille; Koster, Stefan; Sakowski, Erik et al. (2016) Mycobacterium tuberculosis induces the miR-33 locus to reprogram autophagy and host lipid metabolism. Nat Immunol 17:677-86
Tufariello, JoAnn M; Chapman, Jessica R; Kerantzas, Christopher A et al. (2016) Separable roles for Mycobacterium tuberculosis ESX-3 effectors in iron acquisition and virulence. Proc Natl Acad Sci U S A 113:E348-57
Tinaztepe, Emir; Wei, Jun-Rong; Raynowska, Jenelle et al. (2016) Role of Metal-Dependent Regulation of ESX-3 Secretion in Intracellular Survival of Mycobacterium tuberculosis. Infect Immun 84:2255-2263
Vaeth, Martin; Zee, Isabelle; Concepcion, Axel R et al. (2015) Ca2+ Signaling but Not Store-Operated Ca2+ Entry Is Required for the Function of Macrophages and Dendritic Cells. J Immunol 195:1202-17
Sakowski, Erik T; Koster, Stefan; Portal Celhay, Cynthia et al. (2015) Ubiquilin 1 Promotes IFN-?-Induced Xenophagy of Mycobacterium tuberculosis. PLoS Pathog 11:e1005076
Mehra, Alka; Philips, Jennifer A (2014) Analysis of Mycobacterial Protein Secretion. Bio Protoc 4:
Mehra, Alka (2014) Phagolysosomal Trafficking Assay. Bio Protoc 4:

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