This is a competing renewal application of R01 AI51622 entitled "Chemical Mycobacteriology". Mycobacterium tuberculosis (Mtb) infections are difficult to treat owing to the requirement of multiple drugs administered over many months, the emergence of drug-resistant strains, and a complex lifecycle that can include a drug-refractory latent stage. Mtb adapts to diverse environments during disease progression by influencing host cells and altering its own metabolic state. Glycolipids of the outermost capsular layer are thought to contribute to host-pathogen interactions;their underlying biosynthetic machineries might offer new targets for Mtb therapy. Metabolic pathways essential for survival during latency are also attractive drug targets. The broad objectives of this program are (1) to investigate the functions of mycobacterial cell wall glycolipids, and (2) to explore sulfur metabolism as a new niche for drug discovery. During the last granting period we focused our studies on the Mtb-specific trehalose glycolipid sulfolipid-1 (SL-1), a putative Mtb virulence factor. We elucidated the complete genetic and biosynthetic machinery underlying SL-1 and probed its functions in host cells and animals. In the course of these studies we discovered a novel sulfated menaquinone metabolite in Mtb, termed S881, disruption of which produces a hypervirulent phenotype in the mouse infection model. We also validated ATP sulfurylase, the enzyme catalyzing the first committed step in sulfate assimilation, as an attractive drug target. The next granting period will focus on four specific aims, the first two of which builds directly from previous work.
In Aim 1 we will develop small molecule inhibitors of ATP sulfurylase as anti-tuberculosis drugs with possible application toward latent Mtb.
In Aim 2 we will investigate the biosynthesis and function of S881, which we propose to play a role in adaptive respiration. We will also initiate two new research directions.
In Aim 3 we will study trehalose glycolipids in the Mtb relative Mycobacterium marinum, exploiting the zebrafish infection model to profile their expression during infection. Two emerging technologies, MALDI mass spectrometry imaging and metabolic/bioorthogonal labeling for fluorescence imaging, will be employed to probe the dynamics of trehalose glycolipids during the course of disease. Finally, in Aim 4 we will investigate pathways that modulate cell wall structure in response to osmotic stress. A chemical approach to imaging peptidoglycan in vivo will be developed to facilitate these studies.

Public Health Relevance

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is a global pathogen that is thought to infect one-third of the world's population. New treatments for TB are urgently needed, in part because of prevalence of drug-resistant strains and the preponderance of latent TB, which does not respond to any current therapy. This project seeks to understand how the molecules of the mycobacterial cell wall and biosynthetic pathways within the cell contribute to infection. The knowledge from this research will facilitate the development of new anti-TB drugs.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01AI051622-13
Application #
8720665
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Lacourciere, Karen A
Project Start
Project End
Budget Start
Budget End
Support Year
13
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of California Berkeley
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
Berkeley
State
CA
Country
United States
Zip Code
94704
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Meniche, Xavier; Otten, Renee; Siegrist, M Sloan et al. (2014) Subpolar addition of new cell wall is directed by DivIVA in mycobacteria. Proc Natl Acad Sci U S A 111:E3243-51
Siegrist, M Sloan; Bertozzi, Carolyn R (2014) Mycobacterial lipid logic. Cell Host Microbe 15:1-2
Smith, Elizabeth L; Bertozzi, Carolyn R; Beatty, Kimberly E (2014) An expanded set of fluorogenic sulfatase activity probes. Chembiochem 15:1101-5
Beahm, Brendan J; Dehnert, Karen W; Derr, Nicolas L et al. (2014) A visualizable chain-terminating inhibitor of glycosaminoglycan biosynthesis in developing zebrafish. Angew Chem Int Ed Engl 53:3347-52
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Sogi, Kimberly M; Gartner, Zev J; Breidenbach, Mark A et al. (2013) Mycobacterium tuberculosis Rv3406 is a type II alkyl sulfatase capable of sulfate scavenging. PLoS One 8:e65080
Siegrist, M Sloan; Whiteside, Sarah; Jewett, John C et al. (2013) (D)-amino acid chemical reporters reveal peptidoglycan dynamics of an intracellular pathogen. ACS Chem Biol 8:500-5
Seeliger, Jessica C; Holsclaw, Cynthia M; Schelle, Michael W et al. (2012) Elucidation and chemical modulation of sulfolipid-1 biosynthesis in Mycobacterium tuberculosis. J Biol Chem 287:7990-8000
Gilmore, Sarah A; Schelle, Michael W; Holsclaw, Cynthia M et al. (2012) Sulfolipid-1 biosynthesis restricts Mycobacterium tuberculosis growth in human macrophages. ACS Chem Biol 7:863-70

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