Enteropathogenic E. coli (EPEC) are common causes of gastrointestinal illness in very young children in developing nations. These and the related murine pathogen, Citrobacter rodentium, are classified as attaching and effacing (A/E) bacteria because upon ingestion, they intimately attach to and cause effacement of host intestinal cells. A/E bacteria induce shifts in the bacteria groups typically present naturally in the gastrointestinl tract, resulting in dysbiosis and disease. While the molecular basis of A/E bacteria-induced pathology is well studied, the mechanisms that enable these pathogens to successfully compete for resources and drive dysbiosis are not well understood. Additionally, the fitness benefit of intimate adherence to host cells through formation of pedestal-like structures, a feature unique to A/E pathogens, is currently unknown. The long-term objectives of this study are to define the mechanisms that result in pathogen-induced intestinal dysbiosis. Specifically, we aim to 1) illustrate the role of respiration to outgrowth of A/E pathogens during infection and 2) define the mechanisms that govern the fitness benefit provided by C. rodentium intimate attachment to host cells. Our hypothesis states that close adherence to epithelial cells provides A/E pathogens with access to oxygen for respiration, therefore allowing for more efficient energy generation over fermenting commensal microbes. To address our hypothesis, C. rodentium strains will be constructed with disruptions in key respiratory enzymes (bd oxidase and formate dehydrogenase) and attachment genes (pedestal-inducing effector protein EspH) and characterized in vitro with a combination of growth assays, tissue culture assays, and confocal microscopy. Mice will then be orally gavaged with single or mixed inocula for competitive infections to test the benefit derived from respiration and attachment in vivo. Germ-free mouse infections will then allow us to isolate interactions between pathogen and specific members of the microbiota. Quantitative PCR on host and pathogen genes will provide information on the levels of cytokine expression as part of the host response as well as a gauge to judge the metabolic state of C. rodentium under different experimental conditions. Completion of this study will provide valuable information on the mechanisms that result in pathogen-induced gut dysbiosis and a strong foundation on which to develop future strategies to prevent A/E pathogen transmission and disease.

Public Health Relevance

Food-borne pathogenic bacteria contribute significantly to worldwide illness and are of key concern to public health. This project aims to explore how one class of disease-inducing bacteria, one that includes pathogenic E. coli, can invade and successfully colonize the mammalian intestinal tract. By fully understanding the complex interactions between invading pathogens, the normal gut microbes, and the mammalian host we hope to provide the foundation for future therapeutic and preventative strategies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
1F31AI112241-01
Application #
8717328
Study Section
Special Emphasis Panel (ZRG1-F13-C (20))
Program Officer
Adger-Johnson, Diane S
Project Start
2014-02-01
Project End
2017-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
1
Fiscal Year
2014
Total Cost
$35,832
Indirect Cost
Name
University of California Davis
Department
Microbiology/Immun/Virology
Type
Other Domestic Higher Education
DUNS #
047120084
City
Davis
State
CA
Country
United States
Zip Code
95618
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Velazquez, Eric M; Rivera-Chávez, Fabian; Bäumler, Andreas J (2017) Spore Preparation Protocol for Enrichment of Clostridia from Murine Intestine. Bio Protoc 7:
Rivera-Chávez, Fabian; Lopez, Christopher A; Zhang, Lillian F et al. (2016) Energy Taxis toward Host-Derived Nitrate Supports a Salmonella Pathogenicity Island 1-Independent Mechanism of Invasion. MBio 7:
Lopez, Christopher A; Miller, Brittany M; Rivera-Chávez, Fabian et al. (2016) Virulence factors enhance Citrobacter rodentium expansion through aerobic respiration. Science 353:1249-53
Faber, Franziska; Tran, Lisa; Byndloss, Mariana X et al. (2016) Host-mediated sugar oxidation promotes post-antibiotic pathogen expansion. Nature 534:697-9
Rivera-Chávez, Fabian; Zhang, Lillian F; Faber, Franziska et al. (2016) Depletion of Butyrate-Producing Clostridia from the Gut Microbiota Drives an Aerobic Luminal Expansion of Salmonella. Cell Host Microbe 19:443-54
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