Abstract
Enterocytes mediate both nutrient absorption and antimicrobial peptide (AMP) secretion while constantly exposed to osmotic fluctuations. However, the role of volume-regulated anion channel (VRAC), a key osmo-sensitive ion channel, in enterocytes and inflammatory bowel disease (IBD) remains unclear. Here, we show that intestinal epithelial cell (IEC)-specific knockout of LRRC8A, the essential VRAC subunit, exacerbates colitis and inflammation-associated colorectal cancer. VRAC deficiency specifically disrupts enterocyte maturation and zonation, expanding AMP-producing enterocytes while reducing enterocytes responsible for nutrient absorption. Retinoic acid metabolism emerges as the most affected nutrient pathway in VRAC-deficient IECs, with reduced Adh1, Aldh1a2 expression and aldehyde dehydrogenase activity. Supplementation with retinoic acid reversed inflammation caused by VRAC deficiency. Conversely, VRAC deficiency induced gut microbiota dysbiosis, while administration of Lactobacillus species effectively restored microbial balance and alleviated inflammation. This study delineates the role of VRAC in balancing nutrient absorption and antimicrobial defense in enterocytes to safeguard gut homeostasis.
Data availability
Bulk RNA-sequencing data has been deposited in the Genome Sequence Archive with the identifier CRA036214. Single-cell sequencing data has been deposited in the Genome Sequence Archive with the identifier CRA035498. 16S rRNA data has been deposited in Genome Sequence Archive with the identifier CRA038364. Metabolomic data has been deposited in the OMIX, China National Center for Bioinformation / Beijing Institute of Genomics, Chinese Academy of Sciences (https://ngdc.cncb.ac.cn/omix: accession no. OMIX014858; no. OMIX014892; no. OMIX014902). All other data are available within the paper and its supplementary information. Source data are provided with this paper.
References
Neurath, M. F. Current and emerging therapeutic targets for IBD. Nat. Rev. Gastroenterol. Hepatol. 14, 269–278 (2017).
Kobayashi, T. et al. Ulcerative colitis. Nat. Rev. Dis. Prim. 6, 74 (2020).
Peterson, L. W. & Artis, D. Intestinal epithelial cells: regulators of barrier function and immune homeostasis. Nat. Rev. Immunol. 14, 141–153 (2014).
Caruso, R., Lo, B. C. & Núñez, G. Host-microbiota interactions in inflammatory bowel disease. Nat. Rev. Immunol. 20, 411–426 (2020).
Zheng, D., Liwinski, T. & Elinav, E. Interaction between microbiota and immunity in health and disease. Cell Res. 30, 492–506 (2020).
Liu, C. et al. Targeting P2Y14R protects against necroptosis of intestinal epithelial cells through PKA/CREB/RIPK1 axis in ulcerative colitis. Nat. Commun. 15, 2083 (2024).
Massironi, S. et al. Inflammation and malnutrition in inflammatory bowel disease. Lancet Gastroenterol. Hepatol. 8, 579–590 (2023).
Adolph, T. E. et al. The metabolic nature of inflammatory bowel diseases. Nat. Rev. Gastroenterol. Hepatol. 19, 753–767 (2022).
Allaire, J. M. et al. The intestinal epithelium: central coordinator of mucosal immunity. Trends Immunol. 39, 677–696 (2018).
Schwärzler, J., Mayr, L., Grabherr, F., Tilg, H. & Adolph, T. E. Epithelial metabolism as a rheostat for intestinal inflammation and malignancy. Trends Cell Biol. 34, 913–927 (2024).
Moor, A. E. et al. Spatial reconstruction of single enterocytes uncovers broad zonation along the intestinal villus axis. Cell 175, 1156–1167.e15 (2018).
Zwick, R. K. et al. Epithelial zonation along the mouse and human small intestine defines five discrete metabolic domains. Nat. Cell Biol. 26, 250–262 (2024).
Hoffmann, E. K., Lambert, I. H. & Pedersen, S. F. Physiology of cell volume regulation in vertebrates. Physiol. Rev. 89, 193–277 (2009).
Jentsch, T. J. VRACs and other ion channels and transporters in the regulation of cell volume and beyond. Nat. Rev. Mol. Cell Biol. 17, 293–307 (2016).
Voss, F. K. et al. Identification of LRRC8 heteromers as an essential component of the volume-regulated anion channel VRAC. Science 344, 634–638 (2014).
Qiu, Z. et al. SWELL1, a plasma membrane protein, is an essential component of volume-regulated anion channel. Cell 157, 447–458 (2014).
Schober, A. L., Wilson, C. S. & Mongin, A. A. Molecular composition and heterogeneity of the LRRC8-containing swelling-activated osmolyte channels in primary rat astrocytes. J. Physiol. 595, 6939–6951 (2017).
Tucker, B. & Olson, J. E. Glutamate receptor-mediated taurine release from the hippocampus during oxidative stress. J. Biomed. Sci. 17 (Suppl 1), S10 (2010).
Yang, J. et al. Glutamate-releasing SWELL1 channel in astrocytes modulates synaptic transmission and promotes brain damage in stroke. Neuron 102, 813–827.e6 (2019).
Zhou, C. et al. Transfer of cGAMP into bystander cells via LRRC8 volume-regulated anion channels augments STING-mediated interferon responses and anti-viral immunity. Immunity 52, 767–781.e6 (2020).
Lahey, L. J. et al. LRRC8A:C/E heteromeric channels are ubiquitous transporters of cGAMP. Mol. cell 80, 578–591.e5 (2020).
Lutter, D., Ullrich, F., Lueck, J. C., Kempa, S. & Jentsch, T. J. Selective transport of neurotransmitters and modulators by distinct volume-regulated LRRC8 anion channels. J. Cell Sci. 130, 1122–1133 (2017).
Zhou, P., Polovitskaya, M. M. & Jentsch, T. J. LRRC8 N termini influence pore properties and gating of volume-regulated anion channels (VRACs). J. Biol. Chem. 293, 13440–13451 (2018).
Samak, G., Narayanan, D., Jaggar, J. H. & Rao, R. CaV1.3 channels and intracellular calcium mediate osmotic stress-induced N-terminal c-Jun kinase activation and disruption of tight junctions in Caco-2 CELL MONOLAYERS. J. Biol. Chem. 286, 30232–30243 (2011).
Lim, C. H., Bot, A. G. M., de Jonge, H. R. & Tilly, B. C. Osmosignaling and volume regulation in intestinal epithelial cells. Methods Enzymol. 428, 325–342 (2007).
Madison, B. B. et al. Cis elements of the villin gene control expression in restricted domains of the vertical (crypt) and horizontal (duodenum, cecum) axes of the intestine. J. Biol. Chem. 277, 33275–33283 (2002).
Wirtz, S. et al. Chemically induced mouse models of acute and chronic intestinal inflammation. Nat. Protoc. 12, 1295–1309 (2017).
Concepcion, A. R. et al. The volume-regulated anion channel LRRC8C suppresses T cell function by regulating cyclic dinucleotide transport and STING-p53 signaling. Nat. Immunol. 23, 287–302 (2022).
Wu, X. et al. The volume regulated anion channel VRAC regulates NLRP3 inflammasome by modulating itaconate efflux and mitochondria function. Pharmacol. Res. 198, 107016 (2023).
Grivennikov, S. I., Greten, F. R. & Karin, M. Immunity, inflammation, and cancer. Cell 140, 883–899 (2010).
Smillie, C. S. et al. Intra- and inter-cellular rewiring of the human colon during ulcerative colitis. Cell 178, 714–730.e22 (2019).
Planells-Cases, R. et al. Subunit composition of VRAC channels determines substrate specificity and cellular resistance to Pt-based anti-cancer drugs. EMBO J. 34, 2993–3008 (2015).
Nakamura, R. et al. Cryo-EM structure of the volume-regulated anion channel LRRC8D isoform identifies features important for substrate permeation. Commun. Biol. 3, 240 (2020).
López-Cayuqueo, K. I. et al. Renal deletion of LRRC8/VRAC channels induces proximal tubulopathy. J. Am. Soc. Nephrology 33, 1528–1545 (2022).
Berková, L. et al. Terminal differentiation of villus tip enterocytes is governed by distinct Tgfβ superfamily members. EMBO Rep. 24, e56454 (2023).
Lukonin, I. et al. Phenotypic landscape of intestinal organoid regeneration. Nature 586, 275–280 (2020).
Haber, A. L. et al. A single-cell survey of the small intestinal epithelium. Nature 551, 333–339 (2017).
Zhang, T. et al. m6A mRNA modification maintains colonic epithelial cell homeostasis via NF-κB-mediated antiapoptotic pathway. Sci. Adv. 8, eabl5723 (2022).
Beumer, J. & Clevers, H. Cell fate specification and differentiation in the adult mammalian intestine. Nat. Rev. Mol. Cell Biol. 22, 39–53 (2021).
Beumer, J. et al. BMP gradient along the intestinal villus axis controls zonated enterocyte and goblet cell states. Cell Rep. 38, 110438 (2022).
Duester, G. Retinoic acid synthesis and signaling during early organogenesis. Cell 134, 921–931 (2008).
Cao, Y. G. et al. Faecalibaculum rodentium remodels retinoic acid signaling to govern eosinophil-dependent intestinal epithelial homeostasis. Cell Host Microbe 30, 1295–1310.e8 (2022).
Bang, Y.-J. et al. Serum amyloid A delivers retinol to intestinal myeloid cells to promote adaptive immunity. Science 373, eabf9232 (2021).
Mielke, L. A. et al. Retinoic acid expression associates with enhanced IL-22 production by γδ T cells and innate lymphoid cells and attenuation of intestinal inflammation. J. Exp. Med. 210, 1117–1124 (2013).
Mucida, D. et al. Reciprocal TH17 and regulatory T cell differentiation mediated by retinoic acid. Science 317, 256–260 (2007).
Woo, V. et al. Commensal segmented filamentous bacteria-derived retinoic acid primes host defense to intestinal infection. Cell Host Microbe 29, 1744–1756.e5 (2021).
Czarnewski, P. et al. Conserved transcriptomic profile between mouse and human colitis allows unsupervised patient stratification. Nat. Commun. 10, 2892 (2019).
Schlößer, S. et al. Salmonella infection accelerates postnatal maturation of the intestinal epithelium. Proc. Natl. Acad. Sci. USA 122, e2403344122 (2025).
Rosay, T., Jimenez, A. G. & Sperandio, V. Glucuronic acid confers colonization advantage to enteric pathogens. Proc. Natl. Acad. Sci. USA 121, e2400226121 (2024).
Lindemans, C. A. et al. Interleukin-22 promotes intestinal-stem-cell-mediated epithelial regeneration. Nature 528, 560–564 (2015).
Jarade, A. et al. Inflammation triggers ILC3 patrolling of the intestinal barrier. Nat. Immunol. 23, 1317–1323 (2022).
González-Loyola, A. et al. c-MAF coordinates enterocyte zonation and nutrient uptake transcriptional programs. J. Exp. Med. https://doi.org/10.1084/jem.20212418 (2022).
Li, C., Peng, K., Xiao, S., Long, Y. & Yu, Q. The role of Lactobacillus in inflammatory bowel disease: from actualities to prospects. Cell Death Discov. 9, 361 (2023).
Cosovanu, C. et al. Intestinal epithelial c-Maf expression determines enterocyte differentiation and nutrient uptake in mice. J. Exp. Med. https://doi.org/10.1084/jem.20220233 (2022).
Seiler, K. M. et al. Single-cell analysis reveals regional reprogramming during adaptation to massive small bowel resection in mice. Cell. Mol. Gastroenterol. Hepatol. 8, 407–426 (2019).
Hashimoto, T. et al. ACE2 links amino acid malnutrition to microbial ecology and intestinal inflammation. Nature 487, 477–481 (2012).
Jang, K. K. et al. Antimicrobial overproduction sustains intestinal inflammation by inhibiting Enterococcus colonization. Cell Host Microbe 31, 1450–1468.e8 (2023).
Wester, R. A. et al. Retinoic acid signaling drives differentiation toward the absorptive lineage in colorectal cancer. iScience 24, 103444 (2021).
Zhi, Y. et al. Novel DCPIB analogs as dual inhibitors of VRAC/TREK1 channels reduced cGAS-STING mediated interferon responses. Biochem. Pharmacol. 199, 114988 (2022).
Nilius, B. et al. Properties of volume-regulated anion channels in mammalian cells. Prog. Biophys. Mol. Biol. 68, 69–119 (1997).
Jackson, P. S. & Strange, K. Characterization of the voltage-dependent properties of a volume-sensitive anion conductance. J. Gen. Physiol. 105, 661–676 (1995).
Ullrich, F., Reincke, S. M., Voss, F. K., Stauber, T. & Jentsch, T. J. Inactivation and anion selectivity of volume-regulated anion channels (VRACs) depend on C-terminal residues of the first extracellular loop. J. Biol. Chem. 291, 17040–17048 (2016).
Malik, A. et al. SYK-CARD9 signaling axis promotes gut fungi-mediated inflammasome activation to restrict colitis and colon cancer. Immunity 49, 515–530.e5 (2018).
Sato, T. et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459, 262–265 (2009).
Jiang, S. et al. Generic diagramming platform (GDP): a comprehensive database of high-quality biomedical graphics. Nucleic Acids Res. 53, D1670–D1676 (2025).
Acknowledgements
This work was funded by the National Natural Science Foundation of China (82361168640; 82273932; 82574412), Natural Science Foundation of Guangdong Province (2024A1515010561) and Guangzhou Municipal Science and Technology Bureau Guangzhou Key Research and Development Program (2025B03J0074) to P.Z.; Natural Science Foundation of China (82574419) and Major scientific and technological projects of Guangdong Province (2023B1111050008) to S.W.L.; National Natural Science Cross disciplinary Major Research Program (92374203) to G.C.; National Natural Science Foundation of China (82504866) to X.W.; and National Natural Science Foundation of China/Research Grants Council Joint Research Scheme (N_EdUHK205/23) and Guangdong Basic and Applied Basic Research Foundation (2022B151513007) to K.K.L.Y. Figures 4E and 5B were created in BioRender; Figs. 5D, 7A, E, 8B, Figs. S2A, S3A, S7E and S12A were created in BioGDP65.
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Conceptualization, P.Z., K.K.L.Y., and S.W.L.; Investigation, X.Y., S.Z., and X.G.; methodology, X.W., L.M., Y.X., S.L., J.H., Y.Z., and Y.C.; writing—original draft, X.Y. and P.Z.; writing—review & editing, P.Z., K.K.L.Y., X.Y., and S.Z.; funding acquisition, P.Z. and K.K.L.Y.; resources, X.C., F.L., G.C., and S.W.L.; supervision, P.Z., K.K.L.Y., and S.W.L.
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Yi, X., Zhang, S., Gu, X. et al. VRAC coordinates the trade-off between nutrient absorption and antimicrobial defense in enterocytes against inflammation. Nat Commun (2026). https://doi.org/10.1038/s41467-026-69963-3
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DOI: https://doi.org/10.1038/s41467-026-69963-3
