Tuberculosis (TB) remains a major threat to global public health and infection with Mycobacterium tuberculosis (Mtb) is estimated to result in over 2 million deaths annually. Current TB treatments are long and arduous and contribute to the emergence of multi-drug resistant (MDR)- and extensively drug-resistant (XDR)-TB. In order to develop new vaccines and drugs for TB, we need to understand how Mtb evades innate immunity and survives in hostile immune environments. A central feature of Mtb pathogenesis is its ability to grow within macrophages, modulate their activities and interfere with microbicidal functions. We have identified a cell envelope-localized lipoprotein encoding a serine hydrolase, Rv2224c, which is critical for Mtb virulence in vivo, survival in macrophages and resistance to cell envelope-directed stresses. We have named this protein Hip1 (Hydrolase important for pathogenesis 1). We demonstrate that Hip1 suppresses innate immune responses;hip1 mutants induce enhanced proinflammatory responses downstream of Toll-like receptor (TLR)-signaling in macrophages. We hypothesize that Hip1 modifies cell envelope or secreted substrates to modulate innate immunity and host defense. The focus of this application is to understand how Hip1 suppresses host innate immune pathways and determine the molecular and biochemical mechanisms for Hip1 function.
The specific aims are (1) To study the role of Hip1 in suppressing host responses. We will define the TLR-dependent pathways modulated by Hip1 and evaluate whether Hip1-mediated immune suppression promotes Mtb growth in macrophages. We will also determine whether enhanced innate immune responses elicited by the hip1 mutant impact adaptive immunity by studying the priming and functionality of T cell responses. (2) To determine the molecular basis for Hip1 function. First we will characterize the molecular and biochemical basis for Hip1 interaction with a candidate physiological substrate, GroEL2. We will investigate the functional significance of Hip1-dependent cleavage of GroEL2 and its contribution to immune suppression and resisting antimicrobial defense. Second, we will use a panel of synthetic substrates to evaluate the range and specificity of Hip1 hydrolase activity. Finally we will identify additional Hip1 interaction partners and physiological substrates using genetic and biochemical approaches.

Public Health Relevance

The global TB epidemic is fueled by HIV co-infection and the spread of drug resistance, posing a grave threat to public health. The studies outlined here will increase our understanding of an important virulence factor that mediates immune suppression and resistance to host antimicrobial defense. Such proteins are attractive new drug targets for inhibition as they can potentially synergize with antibiotics and simultaneously enhance immune-mediated killing of Mtb.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI083366-03
Application #
8259815
Study Section
Host Interactions with Bacterial Pathogens Study Section (HIBP)
Program Officer
Parker, Tina M
Project Start
2010-05-01
Project End
2015-04-30
Budget Start
2012-05-01
Budget End
2013-04-30
Support Year
3
Fiscal Year
2012
Total Cost
$435,600
Indirect Cost
$188,100
Name
Emory University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
066469933
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Madan-Lala, Ranjna; Sia, Jonathan Kevin; King, Rebecca et al. (2014) Mycobacterium tuberculosis impairs dendritic cell functions through the serine hydrolase Hip1. J Immunol 192:4263-72
Naffin-Olivos, Jacqueline L; Georgieva, Maria; Goldfarb, Nathan et al. (2014) Mycobacterium tuberculosis Hip1 modulates macrophage responses through proteolysis of GroEL2. PLoS Pathog 10:e1004132
Madan-Lala, Ranjna; Peixoto, Katia Vitorello; Re, Fabio et al. (2011) Mycobacterium tuberculosis Hip1 dampens macrophage proinflammatory responses by limiting toll-like receptor 2 activation. Infect Immun 79:4828-38