This is an application for an R21 NIH Exploratory/Developmental Research Grant Award. Our long-term goal is to determine how Mycobacterium tuberculosis (Mtb) disrupts the host immune response to establish infection in humans, and to use this information to develop better vaccines and therapies for tuberculosis (TB). In our prior work, we used proteomics to systematically define the interactions between secreted Mtb proteins and human proteins, and discovered a network of several hundred highly-specific interactions that potentially play important roles in Mtb pathogenesis and host defense. We began an initial genetic analysis of the interaction network, and discovered both a novel virulence factor LpqN, as well as its probable host target, the ubiquitin ligase CBL. The specific goal of this application is to use genetics to probe this biochemically-defined Mtb-host interactome, in order to identify functionally important interactions between Mtb and host proteins.
In Aim 1 we will use bacterial genetics to disrupt the bacterial factors within the interactome, and will test these mutants for growth defects in both mice and isolated ex vivo macrophages.
In Aim 2 we will use CRISPR technology to disrupt the host proteins that interact with those bacterial factors contributing to virulence. We anticipate that genetic analysis of the remaining interactome will identify several bacterial virulence factors, as well as additional host pathways playing important roles in Mtb immunity. These findings will pave the way for future studies that will seek to uncover the molecular mechanisms by which these host factors contribute to immunity, which may suggest ways that the factors can be manipulated for therapeutic benefit.

Public Health Relevance

This project is relevant to human health because understanding how Mycobacterium tuberculosis disrupts the immune system will help lay the foundation for developing future therapies aimed at restoring immune function and thereby improving therapy for tuberculosis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AI140060-02
Application #
9737825
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Lacourciere, Karen A
Project Start
2018-07-05
Project End
2020-06-30
Budget Start
2019-07-01
Budget End
2020-06-30
Support Year
2
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of California Davis
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
047120084
City
Davis
State
CA
Country
United States
Zip Code
95618