Dowdell, A. S.; Roer, R. L.; Bhagavatula, G.; Cartwright, I. M.; Cohen, R. H.; Countess, J. A.; Koch, S. D.; Lee, J. S.; Steiner, C. A.; Thompson, N. T.; Villamaria, Z. F.; Welch, N. M.; Worledge, C. S.; Zhou, L.; Colgan, S. P.

The gastrointestinal tract is home to trillions of microorganisms that interact with their host in profound ways, including regulation of immune, endocrine, and neurological functions. One mechanism by which these microbes interact with their eukaryotic host is through the generation of short-chain fatty acids (SCFAs), which are metabolized by the intestinal epithelium creating a state of \"physiologic hypoxia\". This hypoxia, in turn, results in stabilization and activation of hypoxia-inducible factor (HIF), a transcription factor family shown to support gut barrier function and homeostasis, in the intestinal epithelium. The association between HIF and intestinal homeostasis has been long understood, as both genetic and pharmacologic potentiation of the HIF signaling pathway has been shown to promote barrier function both in vitro and in vivo. Although it has been previously established that pathogenic bacteria regulate HIF stabilization and activity in the intestinal epithelium independent of SCFA metabolism, it is not clear whether this property extends to noninfectious and/or commensal bacterial species. Here, we demonstrate that nonpathogenic, commensal strains of Escherichia coli stabilize HIF in intestinal epithelial cells in vitro. Further, we show that HIF is transcriptionally active in these cells and drives a \"pro-barrier\" transcriptional program. This property was found to be dependent on bacterial aerobic respiration, as genetic elimination of E. coli aerobic respiration abolished HIF stabilization and the subsequent transcriptional phenotype. Finally, we observed induction of tissue hypoxia in vivo using antibiotic-treated mice colonized with wild-type, but not respiration-deficient, E. coli. These findings demonstrate a novel ability for probiotic E. coli to regulate intestinal homeostasis through activation of HIF and suggest that this mechanism might be leveraged in as a novel therapeutic to combat intestinal inflammation, such as that observed during inflammatory bowel disease (IBD).