BEST4⁺ cells: a potential hub of intestinal ion transport and diarrhea manipulation

Abstract

BEST4⁺ cells are a recently identified, functionally specialized intestinal epithelial cell population characterized through single-cell and spatial transcriptomic analyses in humans, pigs, rats, and other vertebrates. These cells characteristically express BEST4, OTOP2, CA7, and GUCY2C, and exhibit particularly high CFTR expression in the small intestine, supporting their roles in luminal pH sensing, fluid-electrolyte homeostasis, and mucus hydration. Emerging evidence from organoid and cross-species studies supports a predominant role for the NOTCH–SPIB signaling axis in driving BEST4⁺ cell differentiation, but their lineage identity and regulatory mechanisms remain incompletely resolved. This review focuses on intestinal epithelial BEST4⁺ cells as a critical cellular hub that connects ion transport physiology with the pathophysiology of secretory diarrhea. We summarize the molecular identity, regional distribution, and cross-species conservation of BEST4⁺ cells. We then evaluate their roles in luminal pH regulation, electrogenic fluid secretion, and mucus barrier integrity. We detail how bacterial enterotoxins activate the GC-C/cGMP/CFTR and cAMP/PKA/CFTR pathways in BEST4⁺ cells to drive pathological fluid hypersecretion, and discuss how viral infections may indirectly engage or amplify BEST4⁺ cell-associated ion-transport pathways. We also outline their relevance to viral diarrhea, inflammatory bowel disease, and cystic fibrosis-associated intestinal dysfunction. Finally, we assess established antidiarrheal approaches and investigational strategies that modulate BEST4⁺ cell-associated pathways, and emphasize the need for cell-type-specific validation in physiologically relevant in vivo models.

Keywords

BEST4⁺ cells / Intestinal epithelium / Ion transport / GC-C/CFTR axis / Secretory diarrhea

References

[1] Abu-Siniyeh A, Khataibeh M, Al-Zyoud W, Al HM. Zebrafish as a model for human epithelial pathology. Lab Anim Res, 2025, 41: 6

[2] Amarachintha S, Harmel-Laws E, Steinbrecher KA. Guanylate cyclase C reduces invasion of intestinal epithelial cells by bacterial pathogens. Sci Rep, 2018, 8 ArticleID: 1521

[3] Ameen NA, Ardito T, Kashgarian M, Marino CR. A unique subset of rat and human intestinal villus cells express the cystic fibrosis transmembrane conductance regulator. Gastroenterology, 1995, 108: 1016-1023

[4] Amiri M, Salari A, Seidler U. Distinct roles of SLC26A3 and CFTR in surface pH regulation and bicarbonate secretion in human intestinal epithelium. Acta Physiol (Oxf), 2026, 242 ArticleID: e70157

[5] Atanga R, Singh V, In JG. Intestinal Enteroendocrine Cells: Present and Future Druggable Targets. Int J Mol Sci, 2023, 24: 8836

[6] Baird AW. Coordinated regulation of colonic fluid and mucus secretion. Front Pharmacol, 2026, 17: 1757501

[7] Barreto e Barreto L, Rattes IC, da Costa AV, Gama P. Paneth cells and their multiple functions. Cell Biol Int, 2022, 46: 701-710

[8] Beumer J, Clevers H. Cell fate specification and differentiation in the adult mammalian intestine. Nat Rev Mol Cell Biol, 2021, 22: 39-53

[9] Bijvelds MJ, Loos M, Bronsveld I, Hellemans A, Bongartz JP, Ver Donck L, et al.. Inhibition of Heat-Stable Toxin-Induced Intestinal Salt and Water Secretion by a Novel Class of Guanylyl Cyclase C Inhibitors. J Infect Dis, 2015, 212: 1806-1815

[10] Bulcaen M, Kortleven P, Liu RB, Maule G, Dreano E, Kelly M, et al.. Prime editing functionally corrects cystic fibrosis-causing CFTR mutations in human organoids and airway epithelial cells. Cell Rep Med, 2024, 5 ArticleID: 101544

[11] Burclaff J, Bliton RJ, Breau KA, Ok MT, Gomez-Martinez I, Ranek JS, et al.. A Proximal-to-Distal Survey of Healthy Adult Human Small Intestine and Colon Epithelium by Single-Cell Transcriptomics. Cell Mol Gastroenterol Hepatol, 2022, 13: 1554-1589

[12] Burton SJ, Hachem C, Abraham JM. Luminal Gastrointestinal Manifestations of Cystic Fibrosis. Curr Gastroenterol Rep, 2021, 23: 4

[13] Cézard JP, Duhamel JF, Meyer M, Pharaon I, Bellaiche M, Maurage C, et al.. Efficacy and tolerability of racecadotril in acute diarrhea in children. Gastroenterology, 2001, 120: 799-805

[14] Chang-Graham AL, Perry JL, Engevik MA, Engevik KA, Scribano FJ, Gebert JT, et al.. Rotavirus induces intercellular calcium waves through ADP signaling. Science, 2020, 370 ArticleID: eabc3621

[15] Chen Y, Wu X, Lai J, Li P, Liu Y, Liu Z, et al.. Single-cell atlas in the living fossil Yangtze sturgeon provides insight into the evolution of fish. Genome Biol, 2025, 26 ArticleID: 277

[16] Choi J, Augenlicht LH. Intestinal stem cells: guardians of homeostasis in health and aging amid environmental challenges. Exp Mol Med, 2024, 56: 495-500

[17] Clevers H. Shifting paradigms in tissue stem cell biology: Insights from the intestine. Cell, 2026, 189: 706-724

[18] Ding S, Song Y, Brulois KF, Pan J, Co JY, Ren L, et al.. Retinoic acid and lymphotoxin signaling promote differentiation of human intestinal M cells. Gastroenterology, 2020, 159: 214-26.e1

[19] Dos Reis DC, Jin J, Santos A, Dastoor P, Muiler C, Zagoren E, et al.. CFTR high expresser BEST4+ cells are pH-sensing neuropod cells: new implications for intestinal physiology and cystic fibrosis disease. Am J Physiol Cell Physiol, 2025, 329: C1411-C1428

[20] Dreano E, Bacchetta M, Simonin J, Galmiche L, Usal C, Slimani L, et al.. Characterization of two rat models of cystic fibrosis-KO and F508del CFTR-generated by CRISPR-Cas9. Anim Model Exp Med, 2019, 2: 297-311

[21] Du N, Zhang K, Guo J, Yan W, Xu L. From mucus secretion to immune surveillance: the evolving roles of goblet cells in intestinal homeostasis and inflammation. Front Immunol, 2026, 17 ArticleID: 1735230

[22] Dupont C, Vernisse B. Anti-diarrheal effects of diosmectite in the treatment of acute diarrhea in children: a review. Paediatr Drugs, 2009, 11: 89-99

[23] Elmentaite R, Kumasaka N, Roberts K, Fleming A, Dann E, King HW, et al.. Cells of the human intestinal tract mapped across space and time. Nature, 2021, 597: 250-255

[24] Enns CB, Harding JCS, Loewen ME. Porcine epidemic diarrhea virus, a member of the Coronaviridae family, increases epithelial secretion in the jejunum by up-regulation of KCNN4. FASEB J, 2018, 32(S1 ArticleID: 750.28

[25] Ettayebi K, Crawford SE, Murakami K, Broughman JR, Karandikar U, Tenge VR, et al.. Replication of human noroviruses in stem cell-derived human enteroids. Science, 2016, 353: 1387-1393

[26] Faizo NL. The intestinal stem cell as a target: a review. Medicine (Baltimore), 2024, 103 ArticleID: e39456

[27] Fang X, Mou Y, Huang Z, Li Y, Han L, Zhang Y, et al.. The sequence and analysis of a Chinese pig genome. Gigascience, 2012, 1 ArticleID: 16

[28] Feng X, Flüchter P, De Tenorio JC, Schneider C. Tuft cells in the intestine, immunity and beyond. Nat Rev Gastroenterol Hepatol, 2024, 21: 852-868

[29] Fischbach W, Andresen V, Eberlin M, Mueck T, Layer P. A Comprehensive Comparison of the Efficacy and Tolerability of Racecadotril with Other Treatments of Acute Diarrhea in Adults. Front Med (Lausanne), 2016, 3: 44

[30] Gan N, Zeng W, Han Y, Chen Q, Jiang Y. Structural mechanism of proton conduction in otopetrin proton channel. Nat Commun, 2024, 15: 7250

[31] Gao X, Yeh HI, Yang Z, Fan C, Jiang F, Howard RJ, et al.. Allosteric inhibition of CFTR gating by CFTRinh-172 binding in the pore. Nat Commun, 2024, 15: 6668

[32] Green KY, Kaufman SS, Nagata BM, Chaimongkol N, Kim DY, Levenson EA, et al.. Human norovirus targets enteroendocrine epithelial cells in the small intestine. Nat Commun, 2020, 11: 2759

[33] Gustafsson JK, Johansson MEV. The role of goblet cells and mucus in intestinal homeostasis. Nat Rev Gastroenterol Hepatol, 2022, 19: 785-803

[34] Gustafsson JK, Lindén SK, Alwan AH, Scholte BJ, Hansson GC, Sjövall H. Carbachol-induced colonic mucus formation requires transport via NKCC1, K⁺ channels and CFTR. Pflugers Arch, 2015, 467: 1403-1415

[35] Hagbom M, Istrate C, Engblom D, Karlsson T, Rodriguez-Diaz J, Buesa J, et al.. Rotavirus stimulates release of serotonin (5-HT) from human enterochromaffin cells and activates brain structures involved in nausea and vomiting. PLoS Pathog, 2011, 7 ArticleID: e1002115

[36] Han X, Mann E, Gilbert S, Guan Y, Steinbrecher KA, Montrose MH, et al.. Loss of guanylyl cyclase C (GCC) signaling leads to dysfunctional intestinal barrier. PLoS ONE, 2011, 6 ArticleID: e16139

[37] Harnik Y, Yakubovsky O, Hoefflin R, Novoselsky R, Bahar Halpern K, Barkai T, et al.. A spatial expression atlas of the adult human proximal small intestine. Nature, 2024, 632: 1101-1109

[38] Hickey JW, Becker WR, Nevins SA, Horning A, Perez AE, Zhu C, et al.. Organization of the human intestine at single-cell resolution. Nature, 2023, 619: 572-584

[39] Højberg O, Canibe N, Poulsen HD, Hedemann MS, Jensen BB. Influence of dietary zinc oxide and copper sulfate on the gastrointestinal ecosystem in newly weaned piglets. Appl Environ Microbiol, 2005, 71: 2267-2277

[40] Hong J, Tallaksen J, Urriola PE. Bridging Nutritional and Environmental Assessment Tools: A One Health Integration Using Zinc Supplementation in Weaned Pigs. Environments, 2025, 12: 279

[41] Huang C, Zhang M, Zheng Q, Yu Q, Yang G, Ren W, et al.. The High-Altitude Adaptation Characteristics of Microbiota-Host Cross-Talk in Yak Gastrointestinal Track. Adv Sci (Weinh), 2026, 13 ArticleID: e14862

[42] Ito G, Okamoto R, Murano T, Shimizu H, Fujii S, Nakata T, et al.. Lineage-specific expression of bestrophin-2 and bestrophin-4 in human intestinal epithelial cells. PLoS ONE, 2013, 8 ArticleID: e79693

[43] Jakab RL, Collaco AM, Ameen NA. Characterization of CFTR high expresser cells in the intestine. Am J Physiol Gastrointest Liver Physiol, 2013, 305: G453-G465

[44] Jones LO, Willms RJ, Xu X, Graham RDV, Eklund M, Shin M, et al.. Single-cell resolution of the adult zebrafish intestine under conventional conditions and in response to an acute Vibrio cholerae infection. Cell Rep, 2023, 42 ArticleID: 113407

[45] Keely SJ, Barrett KE. Intestinal secretory mechanisms and diarrhea. Am J Physiol Gastrointest Liver Physiol, 2022, 322: G405-G420

[46] Kelly M, Trudel S, Brouillard F, Bouillaud F, Colas J, Nguyen-Khoa T, et al.. Cystic fibrosis transmembrane regulator inhibitors CFTR(inh)-172 and GlyH-101 target mitochondrial functions, independently of chloride channel inhibition. J Pharmacol Exp Ther, 2010, 333: 60-69

[47] Kim J, Park H, Park NY, Hwang SI, Kim YE, Sung SI, et al.. Functional maturation of preterm intestinal epithelium through CFTR activation. Commun Biol, 2025, 8: 540

[48] Knoll RL, Brauny MM, Robert E, Cloos L, Waser L, Hilbert K, et al.. CFTR modulator therapy drives microbiome restructuring through improved host physiology in cystic fibrosis: the IMMProveCF phase IV trial. Nat Commun, 2025, 16: 10111

[49] Kolev HM, Kaestner KH. Mammalian Intestinal Development and Differentiation-The State of the Art. Cell Mol Gastroenterol Hepatol, 2023, 16: 809-821

[50] Li H, Wang X, Wang Y, Zhang M, Hong F, Wang H, et al.. Cross-species single-cell transcriptomic analysis reveals divergence of cell composition and functions in mammalian ileum epithelium. Cell Regen, 2022, 11: 19

[51] Ljungholm PL, Ermund A, Söderlund Garsveden MM, Pettersson VL, Gustafsson JK. The anion exchanger slc26a3 regulates colonic mucus expansion during steady state and in response to prostaglandin E2, while Cftr regulates de novo mucus release in response to carbamylcholine. Pflugers Arch, 2024, 476: 1209-1219

[52] Ma T, Thiagarajah JR, Yang H, Sonawane ND, Folli C, Galietta LJ, et al.. Thiazolidinone CFTR inhibitor identified by high-throughput screening blocks cholera toxin-induced intestinal fluid secretion. J Clin Invest, 2002, 110: 1651-1658

[53] Malonga T, Vialaneix N, Beaumont M. BEST4+ cells in the intestinal epithelium. Am J Physiol Cell Physiol, 2024, 326: C1345-C1352

[54] Malonga T, Knudsen C, Alberge J, Lhuillier E, Aymard P, Jones E, et al.. A Single-Cell Atlas of Transcriptome Changes in the Intestinal Epithelium at the Suckling-to-Weaning Transition in Male Rabbits. Cell Mol Gastroenterol Hepatol, 2026, 20 ArticleID: 101628

[55] Manneck D, Manz G, Braun HS, Rosendahl J, Stumpff F. The TRPA1 agonist cinnamaldehyde induces the secretion of HCO3- by the porcine colon. Int J Mol Sci, 2021, 22 ArticleID: 5198

[56] Marasco G, Meacci D, Sarnelli G, Tosetti C, Cremon C, Savarino EV, et al.. Diarrhea management: from pathophysiology to microbiota modulation. Ther Adv Gastroenterol, 2026, 19: 17562848261424324

[57] Melis N, Tauc M, Cougnon M, Bendahhou S, Giuliano S, Rubera I, et al.. Revisiting CFTR inhibition: a comparative study of CFTRinh-172 and GlyH-101 inhibitors. Br J Pharmacol, 2014, 171: 3716-3727

[58] Milenkovic A, Weber BHF. Allele-specific suppression of pathogenic bestrophin-1 transcripts by CRISPR/Cas9-mediated genome editing. Genome Med, 2026, 18 ArticleID: 45

[59] Mitrofanova O, Nikolaev M, Xu Q, Broguiere N, Cubela I, Camp JG, et al.. Bioengineered human colon organoids with in vivo-like cellular complexity and function. Cell Stem Cell, 2024, 31: 1175-1186

[60] Motyka NI, Stewart SR, Porretta CP, Hollifield IE, Bauer DL, Bitoun JP. Enterotoxigenic Escherichia coli Enterotoxins Regulate Epithelial to Immune Relay of IL-33 and IL-1Ra Cytokines. Infect Immun, 2022, 90 ArticleID: e0063721

[61] Muanprasat C, Sonawane ND, Salinas D, Taddei A, Galietta LJ, Verkman AS. Discovery of glycine hydrazide pore-occluding CFTR inhibitors: mechanism, structure-activity analysis, and in vivo efficacy. J Gen Physiol, 2004, 124: 125-137

[62] Mullaney JA, Roy NC, Halliday C, Young W, Altermann E, Kruger MC, et al.. Effects of early postnatal life nutritional interventions on immune-microbiome interactions in the gastrointestinal tract and implications for brain development and function. Front Microbiol, 2022, 13 ArticleID: 960492

[63] Neurath MF, Artis D, Becker C. The intestinal barrier: a pivotal role in health, inflammation, and cancer. Lancet Gastroenterol Hepatol, 2025, 10: 573-592

[64] Nylander O, Sjöblom M, Sedin J, Dahlgren D. Effects of α2-adrenoceptor stimulation on luminal alkalinisation and net fluid flux in rat duodenum. PLoS ONE, 2022, 17 ArticleID: e0273208

[65] Ota H, Suzuki M, Tanabe H, Kusama Y, Seki A, Hara E, et al.. Impaired bicarbonate transport via SLC26A3 and CFTR downregulation promotes mucous cap formation in sessile serrated lesions. Dig Dis Sci, 2026, 71: 941-947

[66] Owji AP, Kittredge A, Zhang Y, Yang T. Structure and function of the bestrophin family of calcium-activated chloride channels. Channels, 2021, 15: 604-623

[67] Owji AP, Wang J, Kittredge A, Clark Z, Zhang Y, Hendrickson WA, et al.. Structures and gating mechanisms of human bestrophin anion channels. Nat Commun, 2022, 13 ArticleID: 3836

[68] Parikh K, Antanaviciute A, Fawkner-Corbett D, Jagielowicz M, Aulicino A, Lagerholm C, et al.. Colonic epithelial cell diversity in health and inflammatory bowel disease. Nature, 2019, 567: 49-55

[69] Pawłowska N, Durda-Masny M, Cofta S, Springer D, Szwed A. Gut dysbiosis driven by CFTR gene mutations in cystic fibrosis patients: from genetic disruption to multisystem consequences and microbiota modulation. Genes (Basel), 2025, 16 ArticleID: 1049

[70] Peters A, Merrington G, Stapleton K, Lofts S. Environmental risk assessment of the use of zinc oxide medicated feeds for weaning piglets in the UK. Environ Sci Adv, 2024, 3: 706-716

[71] Prasad H, Mathew JKK, Visweswariah SS. Receptor Guanylyl Cyclase C and Cyclic GMP in Health and Disease: Perspectives and Therapeutic Opportunities. Front Endocrinol (Lausanne), 2022, 13 ArticleID: 911459

[72] Qiao Y, He C, Xia Y, Ocansey DKW, Mao F. Intestinal mucus barrier: a potential therapeutic target for IBD. Autoimmun Rev, 2025, 24 ArticleID: 103717

[73] Quintero M, Samuelson LC. Paneth Cells: Dispensable yet Irreplaceable for the Intestinal Stem Cell Niche. Cell Mol Gastroenterol Hepatol, 2025, 19 ArticleID: 101443

[74] Read LT, Hahn RW, Thompson CC, Bauer DL, Norton EB, Clements JD. Simultaneous exposure to Escherichia coli heat-labile and heat-stable enterotoxins increases fluid secretion and alters cyclic nucleotide and cytokine production by intestinal epithelial cells. Infect Immun, 2014, 82: 5308-5316

[75] Rebucci R, Staurenghi V, Marchetti L, Giromini C, Bontempo V. Effects of nature identical essential oils (carvacrol, thymol and cinnamaldehyde) on growth performance of piglets and non-invasive markers of antioxidant status and calprotectin release. Livest Sci, 2022, 263 ArticleID: 104959

[76] Schaaf CR, Polkoff KM, Carter A, Stewart AS, Sheahan B, Freund J, et al.. A LGR5 reporter pig model closely resembles human intestine for improved study of stem cells in disease. FASEB J, 2023, 37 ArticleID: e22975

[77] Schiller LR, Santa Ana CA, Morawski SG, Fordtran JS. Mechanism of the antidiarrheal effect of loperamide. Gastroenterology, 1984, 86: 1475-1480

[78] Schiller LR, Santa Ana CA, Morawski SG, Fordtran JS. Studies of the antidiarrheal action of clonidine: effects on motility and intestinal absorption. Gastroenterology, 1985, 89: 982-988

[79] Schuier M, Sies H, Illek B, Fischer H. Cocoa-related flavonoids inhibit CFTR-mediated chloride transport across T84 human colon epithelia. J Nutr, 2005, 135: 2320-2325

[80] Serigado JM, Foulke-Abel J, Hines WC, Hanson JA, In J, Kovbasnjuk O. Ulcerative Colitis: Novel Epithelial Insights Provided by Single Cell RNA Sequencing. Front Med (Lausanne), 2022, 9 ArticleID: 868508

[81] Sheikh A, Tumala B, Vickers TJ, Martin JC, Rosa BA, Sabui S, et al.. Enterotoxigenic Escherichia coli heat-labile toxin drives enteropathic changes in small intestinal epithelia. Nat Commun, 2022, 13: 6886

[82] Sonawane ND, Muanprasat C, Nagatani R, Song Y, Verkman AS. In vivo pharmacology and antidiarrheal efficacy of a thiazolidinone CFTR inhibitor in rodents. J Pharm Sci, 2005, 94: 134-143

[83] Sonawane ND, Hu J, Muanprasat C, Verkman AS. Luminally active, nonabsorbable CFTR inhibitors as potential therapy to reduce intestinal fluid loss in cholera. FASEB J, 2006, 20: 130-132

[84] Song C, Chai Z, Chen S, Zhang H, Zhang X, Zhou Y. Intestinal mucus components and secretion mechanisms: what we do and do not know. Exp Mol Med, 2023, 55: 681-691

[85] Stanforth KJ, Zakhour MI, Chater PI, Wilcox MD, Adamson B, Robson NA, et al.. The MUC2 Gene Product: Polymerisation and Post-Secretory Organisation-Current Models. Polymers (Basel), 2024, 16: 1663

[86] Stas EB, Chen Y, Wolfenden R, Woodworth JC, Goodband RD, Tokach MD, et al.. Effects of butyrate glyceride-based feed ingredient supplementation on weanling pig performance, fecal dry matter, serum chemistry, and intestinal short chain fatty acid concentration. J Anim Sci, 2025, 103 ArticleID: skaf198

[87] Steinhoff MC, Douglas RG, Greenberg HB, Callahan DR. Bismuth subsalicylate therapy of viral gastroenteritis. Gastroenterology, 1980, 78: 1495-1499

[88] Sur A, Segal EX, Nunneley MP, Sinclair JW, Prochaska MK, Dye LE, et al.. Developmental regulation of intestinal BEST4+ cells. bioRxiv, 2025

[89] Tang W, Zhong Y, Wei Y, Deng Z, Mao J, Liu J, et al.. Ileum tissue single-cell mRNA sequencing elucidates the cellular architecture of pathophysiological changes associated with weaning in piglets. BMC Biol, 2022, 20: 123

[90] Tang X, Xiong K, Zeng Y, Fang R. The Mechanism of Zinc Oxide in Alleviating Diarrhea in Piglets after Weaning: A Review from the Perspective of Intestinal Barrier Function. Int J Mol Sci, 2024, 25: 10040

[91] Teng B, Kaplan JP, Liang Z, Krieger Z, Tu YH, Burendei B, et al.. Structural motifs for subtype-specific pH-sensitive gating of vertebrate otopetrin proton channels. Elife, 2022, 11 ArticleID: e77946

[92] Thiagarajah JR, Broadbent T, Hsieh E, Verkman AS. Prevention of toxin-induced intestinal ion and fluid secretion by a small-molecule CFTR inhibitor. Gastroenterology, 2004, 126: 511-519

[93] Thiagarajah JR, Ko EA, Tradtrantip L, Donowitz M, Verkman AS. Discovery and development of antisecretory drugs for treating diarrheal diseases. Clin Gastroenterol Hepatol, 2014, 12: 204-209

[94] Tian X, Michal AM, Li P, Wolfe HR, Waldman SA, Wickstrom E. STa peptide analogs for probing guanylyl cyclase C. Biopolymers, 2008, 90: 713-723

[95] Tradtrantip L, Namkung W, Verkman AS. Crofelemer, an antisecretory antidiarrheal proanthocyanidin oligomer extracted from Croton lechleri, targets two distinct intestinal chloride channels. Mol Pharmacol, 2010, 77: 69-78

[96] Triana S, Stanifer ML, Metz-Zumaran C, Shahraz M, Mukenhirn M, Kee C, et al.. Single-cell transcriptomics reveals immune response of intestinal cell types to viral infection. Mol Syst Biol, 2021, 17 ArticleID: MSB20209833

[97] Troeger H, Loddenkemper C, Schneider T, Schreier E, Epple HJ, Zeitz M, et al.. Structural and functional changes of the duodenum in human norovirus infection. Gut, 2009, 58: 1070-1077

[98] Tuggle KL, Birket SE, Cui X, Hong J, Warren J, Reid L, et al.. Characterization of defects in ion transport and tissue development in cystic fibrosis transmembrane conductance regulator (CFTR)-knockout rats. PLoS ONE, 2014, 9 ArticleID: e91253

[99] Tümmler B, Pallenberg ST, Dittrich AM, Graeber SY, Naehrlich L, Sommerburg O, et al.. Progress of personalized medicine of cystic fibrosis in the times of efficient CFTR modulators. Mol Cell Pediatr, 2025, 12: 6

[100] van Vugt AHM, Bijvelds MJC, de Jonge HR, Meijsen KF, Restin T, Bryant MB, et al.. A potential treatment of congenital sodium diarrhea in patients with activating GUCY2C mutations. Clin Transl Gastroenterol, 2021, 12 ArticleID: e00427

[101] Wakisaka Y, Sugimoto S, Oda M, Pastuhov S, Fujii M, Sato T, et al.. Understanding the Differentiation Pathway to Bestrophin 4-Positive Cells in Human Colonic Epithelium. Gastroenterology, 2025, 169: 154-157

[102] Wang Z, Xia B, Qi S, Zhang X, Zhang X, Li Y, et al.. Bestrophin-4 relays HES4 and interacts with TWIST1 to suppress epithelial-to-mesenchymal transition in colorectal cancer cells. Elife, 2024, 12 ArticleID: RP88879

[103] Wang D, Spoelstra WK, Lin L, Akkerman N, Krueger D, Dayton T, et al.. Interferon-responsive intestinal BEST4/CA7+ cells are targets of bacterial diarrheal toxins. Cell Stem Cell, 2025, 32: 598-612

[104] Wang D, Lim S, van de Wetering WJ, Lopez-Iglesias C, Okura Y, Teranishi-Ikawa Y, et al.. Human gut M cells resemble dendritic cells and present gluten antigen. Nature, 2026, 650: 251-260

[105] Wang Y, Li X, Wang Y, Yang C, Gao X, Zhu K, et al.. DR5 governs compound exocytosis in colonic goblet cells via TATA-box binding protein-dependent bestrophin-2 transcriptional regulation. Adv Sci, 2026, 13 ArticleID: e16789

[106] Warr A, Affara N, Aken B, Beiki H, Bickhart DM, Billis K, et al.. An improved pig reference genome sequence to enable pig genetics and genomics research. Gigascience, 2020, 9 ArticleID: giaa051

[107] Westfall AK, Gopalan SS, Kay JC, Tippetts TS, Cervantes MB, Lackey K, et al.. Single-cell resolution of intestinal regeneration in pythons without crypts illuminates conserved vertebrate regenerative mechanisms. Proc Natl Acad Sci U S A, 2024, 121 ArticleID: e2405463121

[108] Wiarda JE, Becker SR, Sivasankaran SK, Loving CL. Regional epithelial cell diversity in the small intestine of pigs. J Anim Sci, 2023, 101 ArticleID: skac318

[109] Willms RJ, McCabe T, Jones LO, Schade R, Foley E. Rank-dependent control of tuft and BEST4 cell development in the intestine. Nat Commun, 2026

[110] Willms RJ, Foley E. Mechanisms of epithelial growth and development in the zebrafish intestine. Biochem Soc Trans, 2023, 51: 1213-1224

[111] Wu D, Su S, Zhang P, Zha X, Wei Y, Zhang T, et al.. TIGAR regulates intestinal mucus barrier integrity by inhibiting lactylation of G6PD/6PGD in ulcerative colitis. Nat Commun, 2026, 17 ArticleID: 3382

[112] Xia H, Chen H, Cheng X, Yin M, Yao X, Ma J, et al.. Zebrafish: an efficient vertebrate model for understanding role of gut microbiota. Mol Med, 2022, 28: 161

[113] Xiao YY, Zhang Q, Huang F, Rao L, Yao TX, Yang SY, et al.. Single-cell profiling of the pig cecum at various developmental stages. Zool Res, 2024, 45: 55-68

[114] Xiong S, Li T, Chen ZJ, Ma ZC, Lin L, Shi GS, et al.. BEST1-mediated tonic excitation facilitates anxiety-like behaviors in a mouse model of chronic neuropathic pain. Cell Rep, 2026, 45 ArticleID: 117229

[115] Yang N, Garcia MA, Quinton PM. Normal mucus formation requires cAMP-dependent HCO3- secretion and Ca2+-mediated mucin exocytosis. J Physiol, 2013, 591: 4581-4593

[116] Yang L, Wang X, Zhou X, Chen H, Song S, Deng L, et al.. A tunable human intestinal organoid system achieves controlled balance between self-renewal and differentiation. Nat Commun, 2025, 16: 315

[117] Yi H, Wang Z, Yang B, Yang X, Gao K, Xiong Y, et al.. Effects of zinc oxide and condensed tannins on the growth performance and intestinal health of weaned piglets in ETEC-challenged environment. Front Microbiol, 2023, 14: 1181519

[118] Young PG, Levring J, Fiedorczuk K, Blanchard SC, Chen J. Structural basis for CFTR inhibition by CFTRinh-172. Proc Natl Acad Sci U S A, 2024, 121 ArticleID: e2316675121

[119] Yu P, Xie Q, Cai X, Zhong Z, Wei R, Han H, et al.. IPGCA: A Comprehensive Single Cell Atlas of 1 074 127 Porcine Intestinal Cells Revealing Cellular Dynamics, Genetic Regulation, and Cross-Species Conservation. Adv Sci (Weinh), 2025, 12 ArticleID: e07882

[120] Yuan C, Lyu J, Sun X, Wu J, Liu Y. Congenital diarrhea and enteropathies caused by a heterozygous mutation in the GUCY2C gene: a rare case report. Front Pediatr, 2025, 13 ArticleID: 1649272

[121] Zagoren E, Santos AK, Ameen NA, Sumigray K. Generation and manipulation of rat intestinal organoids. J vis Exp, 2023, 196 ArticleID: e65343

[122] Zagoren E, Dias N, Santos AK, Smith ZD, Ameen NA, Sumigray K. Evidence of secondary Notch signaling within the rat small intestine. Development, 2025, 152 ArticleID: dev204277

[123] Zhang W, Sack DA. Recent progress in enterotoxigenic Escherichia coli vaccine research and development. Infect Immun, 2025, 93 ArticleID: e0036825

[124] Zhang J, Zhao D, Yi D, Wu M, Chen H, Wu T, et al.. Microarray analysis reveals the inhibition of intestinal expression of nutrient transporters in piglets infected with porcine epidemic diarrhea virus. Sci Rep, 2019, 9: 19798

[125] Zhang TT, Lei QQ, He J, Guan X, Zhang X, Huang Y, et al.. Bestrophin3 Deficiency in Vascular Smooth Muscle Cells Activates MEKK2/3-MAPK Signaling to Trigger Spontaneous Aortic Dissection. Circulation, 2023, 148: 589-606

[126] Zhang Y, Zhang S, Sun Z, Liu X, Liao G, Niu Z, et al.. Porcine epidemic diarrhea virus causes diarrhea by activating EGFR to regulates NHE3 activity and mobility on plasma membrane. Front Microbiol, 2023, 14 ArticleID: 1237913

[127] Zhao BC, Wang TH, Chen J, Qiu BH, Xu YR, Zhang Q, et al.. Effects of dietary supplementation with a carvacrol-cinnamaldehyde-thymol blend on growth performance and intestinal health of nursery pigs. Porcine Health Manag, 2023, 9: 24

Funding

National Key Research and Development Program of China(2023YFE0124400)

National Natural Science Foundation of China(32573261)

Science and Technology Program of Guangzhou(2024B03J1267)

RIGHTS & PERMISSIONS

The Author(s)