Epithelial-intrinsic nitric oxide synthase 2 sustains host-microbiota dynamics that promote colitis

Hashimoto-Hill, S.; Hunt, B.; Cole, S.; Negi, S.; Waddell, A.; Yang, Z. F.; Wayman, J.; Ollberding, N.; Miraldi, E.; Lim, H.-W.; Vazquez-Torres, A.; Morrison, T.; Denson, L.; Alenghat, T.

doi:10.1101/2025.09.23.677925

bioRxiv · 10.1101/2025.09.23.677925

Epithelial-intrinsic nitric oxide synthase 2 sustains host-microbiota dynamics that promote colitis

Hashimoto-Hill, S.·Hunt, B.·Cole, S.·Negi, S.·Waddell, A.·Yang, Z. F.·Wayman, J.·Ollberding, N.·Miraldi, E.·Lim, H.-W.·Vazquez-Torres, A.·Morrison, T.·Denson, L.·Alenghat, T.

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Abstract

The microbiota influence disease pathogenesis and treatment, however we have limited ability to assess patient status in relation to the microbiota. Here we find that the nitric oxide generating enzyme, nitric oxide synthase 2 (Nos2), is transcriptionally primed in intestinal epithelial cells (IECs), as opposed to immune cells, in inflammatory bowel disease (IBD) patients. Generation of IEC-specific Nos2 knockout mice revealed that epithelial Nos2 activity promoted susceptibility to intestinal disease and sustained a colitogenic microbiota. Epithelial Nos2 increased levels of nitric oxide-derived nitrates and nitrate-metabolizing bacteria in the intestine. Unexpectedly, extra-intestinal nitrates also reflected IEC-intrinsic Nos2 expression, and systemic nitrate concentrations in patients paralleled intestinal Nos2 activation. In fact, temporally inhibiting epithelial Nos2 was sufficient to alter intestinal nitrate homeostasis and inflammation in mice, as well as restrict nitrate production by human intestinal organoids. These data reveal that epithelial nitric oxide metabolism directs host-microbiota dynamics that can alter disease and that monitoring and targeting this axis may benefit patients with IBD.

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Hashimoto-Hill, S., Hunt, B., Cole, S., Negi, S., Waddell, A., Yang, Z. F., Wayman, J., Ollberding, N., Miraldi, E., Lim, H.-W., Vazquez-Torres, A., Morrison, T., Denson, L., Alenghat, T.. 2025-09-25. Epithelial-intrinsic nitric oxide synthase 2 sustains host-microbiota dynamics that promote colitis. https://doi.org/10.1101/2025.09.23.677925

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