Background: The metabolic syndrome encompasses a state of inflammation and metabolic dysfunction, possibly mediated via a disturbed intestinal barrier. Glucagon-like peptide-1 receptor agonists (GLP-1RAs), such as liraglutide, have shown promising anti-inflammatory effects beyond glucose lowering and weight loss, but the underlying mechanism remains to be elucidated. We hypothesised that GLP-1RAs improve the intestinal barrier function and overall inflammatory status by direct gene activation in mucus-secreting Brunner's glands in the mouse duodenum, known for their high density of glucagon-like peptide-1 receptors (GLP-1Rs).
Methods: Using bulk RNA sequencing, in situ hybridisation, and immunohistochemistry, we analysed the change in the genetic phenotype of mouse Brunner's gland cells following GLP-1R activation by liraglutide.
Results: We show that liraglutide induces a novel and robust upregulation of the gene for the Cystic fibrosis transmembrane conductance regulator, Cftr, in Brunner's glands as a part of an overall genetic phenotype involved in ion channel activity, mucus secretion, and hydration via GLP-1R activation. Additionally, we found a robust upregulation of the genes Muc5b, Il33, Ren1, and Vldlr in Brunner's glands.
Conclusion: Collectively, our results imply an enhanced mucus response from Brunner's glands following GLP-1R activation, which might play a role in the effect of GLP-1.
| [1] |
Paone P, Cani PD. Mucus barrier, mucins and gut microbiota: the expected slimy partners?. Gut. 2020; 69(12): 2232-2243.
|
| [2] |
Saklayen MG. The global epidemic of the metabolic syndrome. Curr Hypertens Rep. 2018; 20(2): 12.
|
| [3] |
Cherrington AD, Rajagopalan H, Maggs D, Devière J. Hydrothermal duodenal mucosal resurfacing: role in the treatment of metabolic disease. Gastrointest Endoscopy Clinics N Am. 2017; 27(2): 299-311.
|
| [4] |
Knudsen LB, Lau J. The discovery and development of liraglutide and semaglutide. Front Endocrinol. 2019; 10: 155.
|
| [5] |
Leite AR, Angélico-Gonçalves A, Vasques-Nóvoa F, et al. Effect of glucagon-like peptide-1 receptor agonists on cardiovascular events in overweight or obese adults without diabetes: a meta-analysis of placebo-controlled randomized trials. Diabetes Obes Metab. 2022; 24(8): 1676-1680.
|
| [6] |
Sjöström L, Peltonen M, Jacobson P, et al. Bariatric surgery and long-term cardiovascular events. JAMA. 2012; 307(1): 56-65.
|
| [7] |
Holst JJ. The physiology of glucagon-like peptide 1. Physiol Rev. 2007; 87(4): 1409-1439.
|
| [8] |
Grunddal KV, Jensen EP, Ørskov C, et al. Expression profile of the GLP-1 receptor in the gastrointestinal tract and pancreas in adult female mice. Endocrinology. 2021; 163(1): bqab216.
|
| [9] |
Korner M, Stockli M, Waser B, Reubi JC. GLP-1 receptor expression in human tumors and human normal tissues: potential for in vivo targeting. J Nucl Med. 2007; 48(5): 736-743.
|
| [10] |
Krause WJ. Brunner's glands: a structural, histochemical and pathological profile. Prog Histochem Cytochem. 2000; 35(4): 259-367.
|
| [11] |
Bansil R, Turner BS. The biology of mucus: composition, synthesis and organization. Adv Drug Deliv Rev. 2018; 124: 3-15.
|
| [12] |
Garcia MA, Yang N, Quinton PM. Normal mouse intestinal mucus release requires cystic fibrosis transmembrane regulator-dependent bicarbonate secretion. J Clin Invest. 2009; 119(9): 2613-2622.
|
| [13] |
Roy MG, Livraghi-Butrico A, Fletcher AA, et al. Muc5b is required for airway defence. Nature. 2014; 505(7483): 412-416.
|
| [14] |
Ermund A, Meiss LN, Rodriguez-Pineiro AM, et al. The normal trachea is cleaned by MUC5B mucin bundles from the submucosal glands coated with the MUC5AC mucin. Biochem Biophys Res Commun. 2017; 492(3): 331-337.
|
| [15] |
Collaco AM, Jakab RL, Hoekstra NE, Mitchell KA, Brooks A, Ameen NA. Regulated traffic of anion transporters in mammalian Brunner's glands: a role for water and fluid transport. Am J Physiol Gastrointest Liver Physiol. 2013; 305(3): G258-G275.
|
| [16] |
Johansson ME, Hansson GC. Immunological aspects of intestinal mucus and mucins. Nat Rev Immunol. 2016; 16(10): 639-649.
|
| [17] |
Gustafsson JK, Ermund A, Ambort D, et al. Bicarbonate and functional CFTR channel are required for proper mucin secretion and link cystic fibrosis with its mucus phenotype. J Exp Med. 2012; 209(7): 1263-1272.
|
| [18] |
Bahne E, Sun EWL, Young RL, et al. Metformin-induced glucagon-like peptide-1 secretion contributes to the actions of metformin in type 2 diabetes. JCI Insight. 2018; 3(23): e93936.
|
| [19] |
Voetmann LM, Underwood CR, Rolin B, et al. In vitro cell cultures of Brunner's glands from male mouse to study GLP-1 receptor function. Am J Physiol Cell Physiol. 2022; 322(6): C1260-c1269.
|
| [20] |
Bang-Berthelsen CH, Holm TL, Pyke C, et al. GLP-1 induces barrier protective expression in Brunner's glands and regulates colonic inflammation. Inflamm Bowel Dis. 2016; 22(9): 2078-2097.
|
| [21] |
Chen EYT, Yang N, Quinton PM, Chin W-C. A new role for bicarbonate in mucus formation. Am J Physiol-Lung Cell Mol Physiol. 2010; 299(4): L542-L549.
|
| [22] |
Bang-Berthelsen CH, Holm TL, Pyke C, et al. GLP-1 induces barrier protective expression in Brunner's glands and regulates colonic inflammation. Inflamm Bowel Dis. 2016; 22(9): 2078-2097.
|
| [23] |
Percie du Sert N, Hurst V, Ahluwalia A, et al. The ARRIVE guidelines 2.0: updated guidelines for reporting animal research. PLoS Biol. 2020; 18(7): e3000410.
|
| [24] |
Subramanian A, Tamayo P, Mootha VK, et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci. 2005; 102(43): 15545-15550.
|
| [25] |
Moore BA, Morris GP, Vanner S. A novel in vitro model of Brunner's gland secretion in the guinea pig duodenum. Am J Physiol Gastrointest Liver Physiol. 2000; 278(3): G477-G485.
|
| [26] |
Wright RD, Jennings MA, Florey HW, Lium R. The influence of nerves and drugs on secretion by the small intestine and an investigation of the enzymes in intestinal juice. Quart J expo Physiol. 1940; 30: 70-120.
|
| [27] |
Andersen DB, Grunddal KV, Pedersen J, et al. Using a reporter mouse to map known and novel sites of GLP-1 receptor expression in peripheral tissues of male mice. Endocrinology. 2020; 162(3): bqaa246.
|
| [28] |
Pyke C, Heller RS, Kirk RK, et al. GLP-1 receptor localization in monkey and human tissue: novel distribution revealed with extensively validated monoclonal antibody. Endocrinology. 2014; 155(4): 1280-1290.
|
| [29] |
Garg M, Angus PW, Burrell LM, Herath C, Gibson PR, Lubel JS. Review article: the pathophysiological roles of the renin-angiotensin system in the gastrointestinal tract. Aliment Pharmacol Ther. 2012; 35(4): 414-428.
|
| [30] |
Nishi T, Forgac M. The vacuolar (H+)-ATPases–nature's most versatile proton pumps. Nat Rev Mol Cell Biol. 2002; 3(2): 94-103.
|
| [31] |
Advani A, Kelly DJ, Cox AJ, et al. The (Pro) renin receptor: site-specific and functional linkage to the vacuolar H+-ATPase in the kidney. Hypertension. 2009; 54(2): 261-269.
|
| [32] |
García-Miranda P, Vázquez-Carretero MD, Sesma P, Peral MJ, Ilundain AA. Reelin is involved in the crypt-villus unit homeostasis. Tissue Eng Part A. 2013; 19(1-2): 188-198.
|
| [33] |
Tomkin GH. The intestine as a regulator of cholesterol homeostasis in diabetes. Atheroscler Suppl. 2008; 9(2): 27-32.
|
| [34] |
Alvarez F, Istomine R, Shourian M, et al. The alarmins IL-1 and IL-33 differentially regulate the functional specialisation of Foxp3+ regulatory T cells during mucosal inflammation. Mucosal Immunol. 2019; 12(3): 746-760.
|
| [35] |
Schiering C, Krausgruber T, Chomka A, et al. The alarmin IL-33 promotes regulatory T-cell function in the intestine. Nature. 2014; 513(7519): 564-568.
|
| [36] |
Buzzelli JN, Chalinor HV, Pavlic DI, et al. IL33 is a stomach Alarmin that initiates a skewed Th2 response to injury and infection. Cell Mol Gastroenterol Hepatol. 2015; 1(2): 203-221.e3.
|
| [37] |
Waddell A, Vallance JE, Hummel A, Alenghat T, Rosen MJ. IL-33 induces murine intestinal goblet cell differentiation indirectly via innate lymphoid cell IL-13 secretion. J Immunol. 2019; 202(2): 598-607.
|
| [38] |
Ämmälä C, Drury WJ, 3rdKnerr L, et al. Targeted delivery of antisense oligonucleotides to pancreatic β-cells. Sci Adv. 2018; 4(10): eaat3386.
|
| [39] |
Chassaing B, Koren O, Goodrich JK, et al. Dietary emulsifiers impact the mouse gut microbiota promoting colitis and metabolic syndrome. Nature. 2015; 519(7541): 92-96.
|
| [40] |
Zaki H, Khan S. High-sugar diet predisposes colitis via promoting mucus degrading bacteria in the gut. J Immunol. 2021; 206(1 Supplement): 113.09-.09.
|
| [41] |
Tomas J, Mulet C, Saffarian A, et al. High-fat diet modifies the PPAR-γ pathway leading to disruption of microbial and physiological ecosystem in murine small intestine. Proc Nat Acad Sci USA. 2016; 113(40): E5934-e5943.
|
| [42] |
Ougaard MKE, Kvist PH, Jensen HE, Hess C, Rune I, Sondergaard H. Murine nephrotoxic nephritis as a model of chronic kidney disease. Int J Nephrol. 2018; 2018: 8424502.
|
| [43] |
Tilg H, Zmora N, Adolph TE, Elinav E. The intestinal microbiota fuelling metabolic inflammation. Nat Rev Immunol. 2020; 20(1): 40-54.
|
| [44] |
Cani PD, Amar J, Iglesias MA, et al. Metabolic endotoxemia initiates obesity and insulin resistance. Diabetes. 2007; 56(7): 1761-1772.
|
| [45] |
Everett BM, Donath MY, Pradhan AD, et al. Anti-Inflammatory therapy with canakinumab for the prevention and management of diabetes. J Am Coll Cardiol. 2018; 71(21): 2392-2401.
|
| [46] |
Ridker PM, Everett BM, Thuren T, et al. Antiinflammatory therapy with canakinumab for atherosclerotic disease. N Engl J Med. 2017; 377(12): 1119-1131.
|
| [47] |
Lincoff AM, Brown-Frandsen K, Colhoun HM, et al. Semaglutide and cardiovascular outcomes in obesity without diabetes. N Engl J Med. 2023; 389(24): 2221-2232.
|
| [48] |
Kosiborod MN, Abildstrøm SZ, Borlaug BA, et al. Semaglutide in patients with heart failure with preserved ejection fraction and obesity. N Engl J Med. 2023; 389(12): 1069-1084.
|
| [49] |
Marso SP, Daniels GH, Brown-Frandsen K, et al. Liraglutide and cardiovascular outcomes in type 2 diabetes. N Engl J Med. 2016; 375(4): 311-322.
|
| [50] |
Rodbard HW, Rosenstock J, Canani LH, et al. Oral semaglutide versus empagliflozin in patients with type 2 diabetes uncontrolled on metformin: the PIONEER 2 trial. Diabetes Care. 2019; 42(12): 2272-2281.
|
| [51] |
Wilding JPH, Batterham RL, Davies M, et al. Weight regain and cardiometabolic effects after withdrawal of semaglutide: the STEP 1 trial extension. Diabetes Obes Metab. 2022; 24(8): 1553-1564.
|
| [52] |
Holst JJ, Andersen DB, Grunddal KV. Actions of glucagon-like peptide-1 receptor ligands in the gut. Br J Pharmacol. 2021; 179(4): 727-742. n/a(n/a).
|
| [53] |
Thazhath SS, Marathe CS, Wu T, et al. The glucagon-like peptide 1 receptor agonist exenatide inhibits small intestinal motility, flow, transit, and absorption of glucose in healthy subjects and patients with type 2 diabetes: a randomized controlled trial. Diabetes. 2016; 65(1): 269-275.
|
| [54] |
Wang Z, Saha S, Van Horn S, et al. Gut microbiome differences between metformin- and liraglutide-treated T2DM subjects. Endocrinol, Diab Metab. 2018; 1(1): e00009.
|
| [55] |
Yusta B, Baggio LL, Koehler J, et al. GLP-1R agonists modulate enteric immune responses through the intestinal intraepithelial lymphocyte GLP-1R. Diabetes. 2015; 64(7): 2537-2549.
|
| [56] |
Chen G, Zhang Z, Adebamowo SN, et al. Common and rare exonic MUC5B variants associated with type 2 diabetes in Han Chinese. PLoS One. 2017; 12(3): e0173784.
|
| [57] |
Lisowska A, Wójtowicz J, Walkowiak J. Small intestine bacterial overgrowth is frequent in cystic fibrosis: combined hydrogen and methane measurements are required for its detection. Acta Biochim Pol. 2009; 56(4): 631-634.
|
| [58] |
Hallberg K, Grzegorczyk A, Larson G, Strandvik B. Intestinal permeability in cystic fibrosis in relation to genotype. J Pediatr Gastroenterol Nutr. 1997; 25(3): 290-295.
|
| [59] |
O'Brien S, Mulcahy H, Fenlon H, et al. Intestinal bile acid malabsorption in cystic fibrosis. Gut. 1993; 34(8): 1137-1141.
|
| [60] |
Dalzell AM, Freestone NS, Billington D, Heaf DP. Small intestinal permeability and orocaecal transit time in cystic fibrosis. Arch Dis Child. 1990; 65(6): 585-588.
|
| [61] |
Hendriks HJ, van Kreel B, Forget PP. Effects of therapy with lansoprazole on intestinal permeability and inflammation in young cystic fibrosis patients. J Pediatr Gastroenterol Nutr. 2001; 33(3): 260-265.
|
| [62] |
Reims A, Strandvik B, Sjövall H. Epithelial electrical resistance as a measure of permeability changes in pediatric duodenal biopsies. J Pediatr Gastroenterol Nutr. 2006; 43(5): 619-623.
|
| [63] |
De Lisle RC, Mueller R, Boyd M. Impaired mucosal barrier function in the small intestine of the cystic fibrosis mouse. J Pediatr Gastroenterol Nutr. 2011; 53(4): 371-379.
|
| [64] |
Blotas C, Férec C, Moisan S. Tissue-specific regulation of CFTR gene expression. Int J Mol Sci. 2023; 24(13): 10678.
|
| [65] |
Jonckheere N, Velghe A, Ducourouble MP, Copin MC, Renes IB, Van Seuningen I. The mouse Muc5b mucin gene is transcriptionally regulated by thyroid transcription factor-1 (TTF-1) and GATA-6 transcription factors. Febs j. 2011; 278(2): 282-294.
|
| [66] |
Leung L, Kang J, Rayyan E, et al. Decreased basal chloride secretion and altered cystic fibrosis transmembrane conductance regulatory protein, Villin, GLUT5 protein expression in jejunum from leptin-deficient mice. Diabetes Metab Syndr Obes. 2014; 7: 321-330.
|
| [67] |
Seguella L, Pesce M, Capuano R, et al. High-fat diet impairs duodenal barrier function and elicits glia-dependent changes along the gut-brain axis that are required for anxiogenic and depressive-like behaviors. J Neuroinflam. 2021; 18(1): 115.
|
| [68] |
Takehara K, Tashima K, Takeuchi K. Alterations in duodenal bicarbonate secretion and mucosal susceptibility to acid in diabetic rats. Gastroenterology. 1997; 112(2): 418-428.
|
| [69] |
Cafferata EG, González-Guerrico AM, Giordano L, Pivetta OH, Santa-Coloma TA. Interleukin-1beta regulates CFTR expression in human intestinal T84 cells. Biochim Biophys Acta. 2000; 1500(2): 241-248.
|
| [70] |
Crites KS, Morin G, Orlando V, et al. CFTR knockdown induces proinflammatory changes in intestinal epithelial cells. J Inflamm (Lond). 2015; 12: 62.
|
| [71] |
Allen A, Flemstrom G. Gastroduodenal mucus bicarbonate barrier: protection against acid and pepsin. Am J Physiol Cell Physiol. 2005; 288(1): C1-19.
|
| [72] |
Choi W, Choe S, Lin J, et al. Exendin-4 restores airway mucus homeostasis through the GLP1R-PKA-PPARγ-FOXA2-phosphatase signaling. Mucosal Immunol. 2020; 13(4): 637-651.
|
| [73] |
Nohara H, Nakashima R, Kamei S, et al. Intratracheal GLP-1 receptor agonist treatment up-regulates mucin via p38 and exacerbates emphysematous phenotype in mucus hypersecretory obstructive lung diseases. Biochem Biophys Res Commun. 2020; 524(2): 332-339.
|
| [74] |
Viby NE, Isidor MS, Buggeskov KB, Poulsen SS, Hansen JB, Kissow H. Glucagon-like peptide-1 (GLP-1) reduces mortality and improves lung function in a model of experimental obstructive lung disease in female mice. Endocrinology. 2013; 154(12): 4503-4511.
|
| [75] |
Baekkevold ES, Roussigné M, Yamanaka T, et al. Molecular characterization of NF-HEV, a nuclear factor preferentially expressed in human high endothelial venules. Am J Pathol. 2003; 163(1): 69-79.
|
| [76] |
Bonilla WV, Fröhlich A, Senn K, et al. The alarmin interleukin-33 drives protective antiviral CD8+T cell responses. Science. 2012; 335(6071): 984-989.
|
| [77] |
Schmitz J, Owyang A, Oldham E, et al. IL-33, an interleukin-1-like cytokine that signals via the IL-1 receptor-related protein ST2 and induces T helper type 2-associated cytokines. Immunity. 2005; 23(5): 479-490.
|
| [78] |
Pichery M, Mirey E, Mercier P, et al. Endogenous IL-33 is highly expressed in mouse epithelial barrier tissues, lymphoid organs, brain, embryos, and inflamed tissues: in situ analysis using a novel Il-33–LacZ gene trap reporter strain. J Immunol. 2012; 188(7): 3488-3495.
|
| [79] |
Shao D, Perros F, Caramori G, et al. Nuclear IL-33 regulates soluble ST2 receptor and IL-6 expression in primary human arterial endothelial cells and is decreased in idiopathic pulmonary arterial hypertension. Biochem Biophys Res Commun. 2014; 451(1): 8-14.
|
| [80] |
Ali S, Mohs A, Thomas M, et al. The dual function cytokine IL-33 interacts with the transcription factor NF-κB to dampen NF-κB–stimulated gene transcription. J Immunol. 2011; 187(4): 1609-1616.
|
| [81] |
Fu AKY, Hung K-W, Yuen MYF, et al. IL-33 ameliorates Alzheimer's disease-like pathology and cognitive decline. Proc Natl Acad Sci. 2016; 113(19): E2705-E2713.
|
| [82] |
Perman JC, Boström P, Lindbom M, et al. The VLDL receptor promotes lipotoxicity and increases mortality in mice following an acute myocardial infarction. J Clin Invest. 2011; 121(7): 2625-2640.
|
| [83] |
Alexander A, Herz J, Calvier L. Reelin through the years: from brain development to inflammation. Cell Rep. 2023; 42(6): 112669.
|
| [84] |
Kounatidis D, Vallianou NG, Poulaki A, et al. ApoB100 and atherosclerosis: what's new in the 21st century? Metabolites. 2024; 14(2): 123.
|
| [85] |
Huang JK, Lee HC. Emerging evidence of pathological roles of very-low-density lipoprotein (VLDL). Int J Mol Sci. 2022; 23(8): 4300.
|
| [86] |
Wang N, Merits A, Veit M, et al. LDL receptor in alphavirus entry: structural analysis and implications for antiviral therapy. Nat Commun. 2024; 15(1): 4906.
|
| [87] |
Holm I, Ollo R, Panthier JJ, Rougeon F. Evolution of aspartyl proteases by gene duplication: the mouse renin gene is organized in two homologous clusters of four exons. Embo j. 1984; 3(3): 557-562.
|
| [88] |
Takahashi N, Lopez ML. Ren1c homozygous null mice are hypotensive and polyuric, but heterozygotes are indistinguishable from wild-type. J Am Soc Nephrol. 2005; 16(1): 125-132.
|
| [89] |
Vara E, Arias-Díaz J, Garcia C, Balibrea JL, Blázquez E. Glucagon-like peptide-1(7-36) amide stimulates surfactant secretion in human type II pneumocytes. Am J Respir Crit Care Med. 2001; 163(4): 840-846.
|
| [90] |
Voetmann LM, Rolin B, Kirk RK, Pyke C, Hansen AK. The intestinal permeability marker FITC-dextran 4 kDa should be dosed according to lean body mass in obese mice. Nutr Diab. 2023; 13(1): 1.
|
| [91] |
Zhao R, Watt AJ, Li J, et al. GATA6 is essential for embryonic development of the liver but dispensable for early heart formation. Mol Cell Biol. 2005; 25(7): 2622-2631.
|
| [92] |
Colledge WH, Abella BS, Southern KW, et al. Generation and characterization of a delta F508 cystic fibrosis mouse model. Nat Genet. 1995; 10(4): 445-452.
|
| [93] |
Wang F, Flanagan J, Su N, et al. RNAscope: a novel in situ RNA analysis platform for formalin-fixed, paraffin-embedded tissues. J Mol Diagn. 2012; 14(1): 22-29.
|
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