Gut Microbial Trimethylamine is Elevated in Alcohol-Associated Hepatitis and Contributes to Ethanol-Induced Liver Injury in Mice

Helsley, R. N.; Miyata, T.; Kadam, A.; Venkateshwari, V.; Sangwan, N.; Huang, E. C.; Banerjee, R.; Brown, A. L.; Fung, K. K.; Massey, W.; Neumann, C.; Danny, O.; Osborn, L. J.; Schugar, R. C.; McMullen, M. R.; Bellar, A.; Poulsen, K. L.; Kim, A.; Pathak, V.; Mrdjen, M.; Anderson, J. T.; Willard, B.; McClain, C. J.; Mitchell, M.; McCullough, A. J.; Radaeva, S.; Barton, B.; Szabo, G.; Dasarathy, S.; Garcia-Garcia, J. C.; Rotroff, D. M.; Allende, D. S.; Wang, Z.; Hazen, S. L.; Nagy, L. E.; Brown, J. M.

doi:10.1101/2021.12.31.474633

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bioRxiv · 10.1101/2021.12.31.474633

Gut Microbial Trimethylamine is Elevated in Alcohol-Associated Hepatitis and Contributes to Ethanol-Induced Liver Injury in Mice

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Abstract

BACKGROUNDThere is mounting evidence that microbes resident in the human intestine contribute to diverse alcohol-associated liver diseases (ALD) including the most deadly form known as alcohol-associated hepatitis (AH). However, mechanisms by which gut microbes synergize with excessive alcohol intake to promote liver injury are poorly understood. Furthermore, whether drugs that selectively target gut microbial metabolism can improve ALD has never been tested. METHODSWe used liquid chromatography tandem mass spectrometry to quantify the levels of microbe and host choline co-metabolites in healthy controls and AH patients, finding elevated levels of the microbial metabolite trimethylamine (TMA) in AH. In subsequent studies, we treated mice with non-lethal bacterial choline TMA lyase (CutC/D) inhibitors to blunt gut microbe-dependent production of TMA in the context of chronic ethanol administration. Indices of liver injury were quantified by complementary RNA sequencing, biochemical, and histological approaches. In addition, we examined the impact of ethanol consumption and TMA lyase inhibition on gut microbiome structure via 16S rRNA sequencing. RESULTSWe show the gut microbial choline metabolite trimethylamine (TMA) is elevated in AH patients and correlates with reduced hepatic expression of the TMA oxygenase flavin-containing monooxygenase 3 (FMO3). Provocatively, we find that small molecule inhibition of gut microbial CutC/D activity protects mice from ethanol-induced liver injury. CutC/D inhibitor-driven improvement in ethanol-induced liver injury is associated with distinct reorganization of the gut microbiome and host liver transcriptome. CONCLUSIONSThe microbial metabolite TMA is elevated in patients with AH, and inhibition of TMA production from gut microbes can protect mice from ethanol-induced liver injury.

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Helsley, R. N., Miyata, T., Kadam, A., Venkateshwari, V., Sangwan, N., Huang, E. C., Banerjee, R., Brown, A. L., Fung, K. K., Massey, W., Neumann, C., Danny, O., Osborn, L. J., Schugar, R. C., McMullen, M. R., Bellar, A., Poulsen, K. L., Kim, A., Pathak, V., Mrdjen, M., Anderson, J. T., Willard, B., McClain, C. J., Mitchell, M., McCullough, A. J., Radaeva, S., Barton, B., Szabo, G., Dasarathy, S., Garcia-Garcia, J. C., Rotroff, D. M., Allende, D. S., Wang, Z., Hazen, S. L., Nagy, L. E., Brown, J. M.. 2022-01-01. Gut Microbial Trimethylamine is Elevated in Alcohol-Associated Hepatitis and Contributes to Ethanol-Induced Liver Injury in Mice. https://doi.org/10.1101/2021.12.31.474633

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