TLR4 signals through islet macrophages to alter cytokine secretion during diabetes

in Journal of Endocrinology
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  • 1 Lee Kong Chian School of Medicine, Nanyang Technological University Singapore, Singapore, Singapore
  • 2 Singapore Eye Research Institute (SERI), Singapore General Hospital, Singapore, Singapore

Correspondence should be addressed to Y Ali: yusuf.ali@ntu.edu.sg

*(Y Ali is the lead author)

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Toll-like receptors (TLRs), particularly TLR4, may act as immune sensors for metabolic stress signals such as lipids and link tissue metabolic changes to innate immunity. TLR signalling is not only tissue-dependent but also cell-type dependent and recent studies suggest that TLRs are not restricted to innate immune cells alone. Pancreatic islets, a hub of metabolic hormones and cytokines, respond to TLR signalling. However, the source of TLR signalling within the islet remain poorly understood. Uncovering the specific cell source and its role in mediating TLR signalling, especially within type 2 diabetes (T2D) islet will yield new targets to tackle islet inflammation, hormone secretion dysregulation and ultimately diabetes. In the present study, we immuno-characterised TLRs linked to pancreatic islets in both healthy and obese diabetic mice. We found that while TLRs1–4 and TLR9 were expressed in mouse islets, these TLRs did not co-localise with insulin-producing β-cells. β-Cells from obese diabetic mice were also devoid of these TLRs. While TLR immunoreactivity in obese mice islets increased, this was driven mostly by increased islet endothelial cell and islet macrophage presence. Analysis of human islet single-cell RNA-seq databases revealed that macrophages were an important source of islet TLRs. However, only TLR4 and TLR8 showed variation and cell-type specificity in their expression patterns. Cell depletion experiments in isolated mouse islets showed that TLR4 signalled through macrophages to alter islet cytokine secretome. Together, these studies suggest that islet macrophages are a dominant source of TLR4-mediated signalling in both healthy and diabetic islets.

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  • Akira S, Takeda K & Kaisho T 2001 Toll-like receptors: critical proteins linking innate and acquired immunity. Nature Immunology 2 675680. (https://doi.org/10.1038/90609)

    • Search Google Scholar
    • Export Citation
  • Baskin DG 2015 A historical perspective on the identification of cell types in pancreatic islets of Langerhans by staining and histochemical techniques. Journal of Histochemistry and Cytochemistry 63 543558. (https://doi.org/10.1369/0022155415589119)

    • Search Google Scholar
    • Export Citation
  • Brissova M, Shostak A, Shiota M, Wiebe PO, Poffenberger G, Kantz J, Chen Z, Carr C, Jerome WG, Chen J, 2006 Pancreatic islet production of vascular endothelial growth factor – a is essential for islet vascularization, revascularization, and function. Diabetes 55 29742985. (https://doi.org/10.2337/db06-0690)

    • Search Google Scholar
    • Export Citation
  • Burrows MP, Volchkov P, Kobayashi KS & Chervonsky AV 2015 Microbiota regulates type 1 diabetes through toll-like receptors. PNAS 112 99739977. (https://doi.org/10.1073/pnas.1508740112)

    • Search Google Scholar
    • Export Citation
  • Carrero JA, Mccarthy DP, Ferris ST, Wan X, Hu H, Zinselmeyer BH, Vomund AN & Unanue ER 2017 Resident macrophages of pancreatic islets have a seminal role in the initiation of autoimmune diabetes of NOD mice. PNAS 114 E10418E10427. (https://doi.org/10.1073/pnas.1713543114)

    • Search Google Scholar
    • Export Citation
  • Carter JD, Dula SB, Corbin KL, Wu R & Nunemaker CS 2009 A practical guide to rodent islet isolation and assessment. Biological Procedures Online 11 331. (https://doi.org/10.1007/s12575-009-9021-0)

    • Search Google Scholar
    • Export Citation
  • Chittezhath M, Gunaseelan D, Zheng X, Hasan R, Tay VSY, Lim ST, Wang X, Berggren PO, Bornstein S, Boehm B, 2019 Islet macrophages are associated with islet vascular remodeling and compensatory hyperinsulinemia during diabetes. American Journal of Physiology: Endocrinology and Metabolism 317 E1108E1120. (https://doi.org/10.1152/ajpendo.00248.2019)

    • Search Google Scholar
    • Export Citation
  • Cucak H, Mayer C, Tonnesen M, Thomsen LH, Grunnet LG & Rosendahl A 2014 Macrophage contact dependent and independent TLR4 mechanisms induce beta-cell dysfunction and apoptosis in a mouse model of type 2 diabetes. PLoS ONE 9 e90685. (https://doi.org/10.1371/journal.pone.0090685)

    • Search Google Scholar
    • Export Citation
  • Dasu MR, Devaraj S, Park S & Jialal I 2010 Increased toll-like receptor (TLR) activation and TLR ligands in recently diagnosed type 2 diabetic subjects. Diabetes Care 33 861868. (https://doi.org/10.2337/dc09-1799)

    • Search Google Scholar
    • Export Citation
  • De Groot D, Hoefer IE, Grundmann S, Schoneveld A, Haverslag RT, Van Keulen JK, Bot PT, Timmers L, Piek JJ, Pasterkamp G, 2011 Arteriogenesis requires toll-like receptor 2 and 4 expression in bone-marrow derived cells. Journal of Molecular and Cellular Cardiology 50 2532. (https://doi.org/10.1016/j.yjmcc.2010.08.006)

    • Search Google Scholar
    • Export Citation
  • Devaraj S, Dasu MR, Rockwood J, Winter W, Griffen SC & Jialal I 2008 Increased toll-like receptor (TLR) 2 and TLR4 expression in monocytes from patients with type 1 diabetes: further evidence of a proinflammatory state. Journal of Clinical Endocrinology and Metabolism 93 578583. (https://doi.org/10.1210/jc.2007-2185)

    • Search Google Scholar
    • Export Citation
  • Devaraj S, Jialal I, Yun JM & Bremer A 2011 Demonstration of increased toll-like receptor 2 and toll-like receptor 4 expression in monocytes of type 1 diabetes mellitus patients with microvascular complications. Metabolism: Clinical and Experimental 60 256259. (https://doi.org/10.1016/j.metabol.2010.01.005)

    • Search Google Scholar
    • Export Citation
  • Dogusan Z, Garcia M, Flamez D, Alexopoulou L, Goldman M, Gysemans C, Mathieu C, Libert C, Eizirik DL & Rasschaert J 2008 Double-stranded RNA induces pancreatic beta-cell apoptosis by activation of the toll-like receptor 3 and interferon regulatory factor 3 pathways. Diabetes 57 12361245. (https://doi.org/10.2337/db07-0844)

    • Search Google Scholar
    • Export Citation
  • Donath MY 2014 Targeting inflammation in the treatment of type 2 diabetes: time to start. Nature Reviews: Drug Discovery 13 465476. (https://doi.org/10.1038/nrd4275)

    • Search Google Scholar
    • Export Citation
  • Garay-Malpartida HM, Mourao RF, Mantovani M, Santos IA, Sogayar MC & Goldberg AC 2011 Toll-like receptor 4 (TLR4) expression in human And murine pancreatic beta-cells affects cell viability and insulin homeostasis. BMC Immunology 12 18. (https://doi.org/10.1186/1471-2172-12-18)

    • Search Google Scholar
    • Export Citation
  • Goldberg A, Parolini M, Chin BY, Czismadia E, Otterbein LE, Bach FH & Wang H 2007 Toll-like receptor 4 suppression leads to islet allograft survival. FASEB Journal 21 28402848. (https://doi.org/10.1096/fj.06-7910com)

    • Search Google Scholar
    • Export Citation
  • Grun D, Muraro MJ, Boisset JC, Wiebrands K, Lyubimova A, Dharmadhikari G, Van Den Born M, Van Es J, Jansen E, Clevers H, 2016 De novo prediction of stem cell identity using single-cell transcriptome data. Cell Stem Cell 19 266277. (https://doi.org/10.1016/j.stem.2016.05.010)

    • Search Google Scholar
    • Export Citation
  • Ha H, Lee JH, Kim HN, Kwak HB, Kim HM, Lee SE, Rhee JH, Kim HH & Lee ZH 2008 Stimulation by TLR5 modulates osteoclast differentiation through STAT1/IFN-beta. Journal of Immunology 180 13821389. (https://doi.org/10.4049/jimmunol.180.3.1382)

    • Search Google Scholar
    • Export Citation
  • He W, Yuan T, Choezom D, Hunkler H, Annamalai K, Lupse B & Maedler K 2018 Ageing potentiates diet-induced glucose intolerance, beta-cell failure and tissue inflammation through TLR4. Scientific Reports 8 2767. (https://doi.org/10.1038/s41598-018-20909-w)

    • Search Google Scholar
    • Export Citation
  • He W, Rebello O, Savino R, Terracciano R, Schuster-Klein C, Guardiola B & Maedler K 2019 TLR4 triggered complex inflammation in human pancreatic islets. Biochimica et Biophysica Acta: Molecular Basis of Disease 1865 8697. (https://doi.org/10.1016/j.bbadis.2018.09.030)

    • Search Google Scholar
    • Export Citation
  • Helminen O, Huhta H, Kauppila JH, Lehenkari PP, Saarnio J & Karttunen TJ 2017 Localization of nucleic acid-sensing toll-like receptors in human and mouse pancreas. APMIS 125 8592. (https://doi.org/10.1111/apm.12632)

    • Search Google Scholar
    • Export Citation
  • Himes RW & Smith CW 2010 Tlr2 is critical for diet-induced metabolic syndrome in a murine model. FASEB Journal 24 731739. (https://doi.org/10.1096/fj.09-141929)

    • Search Google Scholar
    • Export Citation
  • Ilvesaro JM, Merrell MA, Li L, Wakchoure S, Graves D, Brooks S, Rahko E, Jukkola-Vuorinen A, Vuopala KS, Harris KW, 2008 Toll-like receptor 9 mediates CpG oligonucleotide-induced cellular invasion. Molecular Cancer Research 6 15341543. (https://doi.org/10.1158/1541-7786.MCR-07-2005)

    • Search Google Scholar
    • Export Citation
  • Ji Y, Sun S, Shrestha N, Darragh LB, Shirakawa J, Xing Y, He Y, Carboneau BA, Kim H, An D, 2019 Toll-like receptors TLR2 and TLR4 block the replication of pancreatic beta cells in diet-induced obesity. Nature Immunology 20 677686. (https://doi.org/10.1038/s41590-019-0396-z)

    • Search Google Scholar
    • Export Citation
  • Jia L, Vianna CR, Fukuda M, Berglund ED, Liu C, Tao C, Sun K, Liu T, Harper MJ, Lee CE, 2014 Hepatocyte toll-like receptor 4 regulates obesity-induced inflammation and insulin resistance. Nature Communications 5 3878. (https://doi.org/10.1038/ncomms4878)

    • Search Google Scholar
    • Export Citation
  • Kim F, Pham M, Luttrell I, Bannerman DD, Tupper J, Thaler J, Hawn TR, Raines EW & Schwartz MW 2007 Toll-like receptor-4 mediates vascular inflammation and insulin resistance in diet-induced obesity. Circulation Research 100 15891596. (https://doi.org/10.1161/CIRCRESAHA.106.142851)

    • Search Google Scholar
    • Export Citation
  • Konner AC & Bruning JC 2011 Toll-Like receptors: linking inflammation to metabolism. Trends in Endocrinology and Metabolism 22 1623. (https://doi.org/10.1016/j.tem.2010.08.007)

    • Search Google Scholar
    • Export Citation
  • Kraakman MJ, Murphy AJ, Jandeleit-Dahm K & Kammoun HL 2014 Macrophage polarization in obesity and type 2 diabetes: weighing down our understanding of macrophage function? Frontiers in Immunology 5 470. (https://doi.org/10.3389/fimmu.2014.00470)

    • Search Google Scholar
    • Export Citation
  • Krüger B, Yin N, Zhang N, Yadav A, Coward W, Lal G, Zang W, S Heeger P, Bromberg JS, Murphy B, 2010 Islet-expressed TLR2 and TLR4 sense injury and mediate early graft failure after transplantation. European Journal of Immunology 40 29142924. (https://doi.org/10.1002/eji.201040601)

    • Search Google Scholar
    • Export Citation
  • Lawlor N, George J, Bolisetty M, Kursawe R, Sun L, Sivakamasundari V, Kycia I, Robson P & Stitzel ML 2017 Single-cell transcriptomes identify human islet cell signatures and reveal cell-type-specific expression changes in type 2 diabetes. Genome Research 27 208222. (https://doi.org/10.1101/gr.212720.116)

    • Search Google Scholar
    • Export Citation
  • Lebreton F, Berishvili E, Parnaud G, Rouget C, Bosco D, Berney T & Lavallard V 2018 NLRP3 inflammasome is expressed and regulated in human islets. Cell Death and Disease 9 726. (https://doi.org/10.1038/s41419-018-0764-x)

    • Search Google Scholar
    • Export Citation
  • Lee MS, Kim DH, Lee JC, Kim S & Kim HS 2011 Role of TLR2 in the pathogenesis of autoimmune diabetes and its therapeutic implication. Diabetes/Metabolism Research and Reviews 27 797801. (https://doi.org/10.1002/dmrr.1231)

    • Search Google Scholar
    • Export Citation
  • Li H & Sun B 2007 Toll-like receptor 4 in atherosclerosis. Journal of Cellular and Molecular Medicine 11 8895. (https://doi.org/10.1111/j.1582-4934.2007.00011.x)

    • Search Google Scholar
    • Export Citation
  • Li J, Jin C, Cleveland JC, Ao L, Xu D, Fullerton DA & Meng X 2010 Enhanced inflammatory responses to toll-like receptor 2/4 stimulation in type 1 diabetic coronary artery endothelial cells: the effect of insulin. Cardiovascular Diabetology 9 90. (https://doi.org/10.1186/1475-2840-9-90)

    • Search Google Scholar
    • Export Citation
  • Li M, Song L, Gao X, Chang W & Qin X 2012 Toll-like receptor 4 on islet beta cells senses expression changes in high-mobility group box 1 and contributes to the initiation of type 1 diabetes. Experimental and Molecular Medicine 44 260267. (https://doi.org/10.3858/emm.2012.44.4.021)

    • Search Google Scholar
    • Export Citation
  • Li J, Chen L, Zhang Y, Zhang WJ, Xu W, Qin Y, Xu J & Zou D 2013 TLR4 is required for the obesity-induced pancreatic beta cell dysfunction. Acta Biochimica et Biophysica Sinica 45 10301038. (https://doi.org/10.1093/abbs/gmt092)

    • Search Google Scholar
    • Export Citation
  • Masters SL, Dunne A, Subramanian SL, Hull RL, Tannahill GM, Sharp FA, Becker C, Franchi L, Yoshihara E, Chen Z, 2010 Activation of the NLRP3 inflammasome by islet amyloid polypeptide provides a mechanism for enhanced IL-1beta in type 2 diabetes. Nature Immunology 11 897904. (https://doi.org/10.1038/ni.1935)

    • Search Google Scholar
    • Export Citation
  • Muraro MJ, Dharmadhikari G, Grun D, Groen N, Dielen T, Jansen E, Van Gurp L, Engelse MA, Carlotti F, De Koning EJ, 2016 A single-cell transcriptome atlas of the human pancreas. Cell Systems 3 385.e3394.e3. (https://doi.org/10.1016/j.cels.2016.09.002)

    • Search Google Scholar
    • Export Citation
  • Nackiewicz D, Dan M, He W, Kim R, Salmi A, Rutti S, Westwell-Roper C, Cunningham A, Speck M, Schuster-Klein C, 2014 TLR2/6 and TLR4-activated macrophages contribute to islet inflammation and impair beta cell insulin gene expression via IL-1 and IL-6. Diabetologia 57 16451654. (https://doi.org/10.1007/s00125-014-3249-1)

    • Search Google Scholar
    • Export Citation
  • Nackiewicz D, Dan M, Speck M, Chow SZ, Chen YC, Pospisilik JA, Verchere CB & Ehses JA 2020 Islet macrophages shift to a reparative state following pancreatic beta-cell death and are a major source of islet insulin-like growth factor-1. iScience 23 100775. (https://doi.org/10.1016/j.isci.2019.100775)

    • Search Google Scholar
    • Export Citation
  • Nyqvist D, Kohler M, Wahlstedt H & Berggren PO 2005 Donor islet endothelial cells participate in formation of functional vessels within pancreatic islet grafts. Diabetes 54 22872293. (https://doi.org/10.2337/diabetes.54.8.2287)

    • Search Google Scholar
    • Export Citation
  • Nyqvist D, Speier S, Rodriguez-Diaz R, Molano RD, Lipovsek S, Rupnik M, Dicker A, Ilegems E, Zahr-Akrawi E, Molina J, 2011 Donor islet endothelial cells in pancreatic islet revascularization. Diabetes 60 25712577. (https://doi.org/10.2337/db10-1711)

    • Search Google Scholar
    • Export Citation
  • Rasschaert J, Ladriere L, Urbain M, Dogusan Z, Katabua B, Sato S, Akira S, Gysemans C, Mathieu C & Eizirik DL 2005 Toll-like receptor 3 and STAT-1 contribute to double-stranded RNA+ interferon-gamma-induced apoptosis in primary pancreatic beta-cells. Journal of Biological Chemistry 280 3398433991. (https://doi.org/10.1074/jbc.M502213200)

    • Search Google Scholar
    • Export Citation
  • Razolli DS, Moraes JC, Morari J, Moura RF, Vinolo MA & Velloso LA 2015 TLR4 expression in bone marrow-derived cells is both necessary and sufficient to produce the insulin resistance phenotype in diet-induced obesity. Endocrinology 156 103113. (https://doi.org/10.1210/en.2014-1552)

    • Search Google Scholar
    • Export Citation
  • Regel I, Raulefs S, Benitz S, Mihaljevic C, Rieder S, Leinenkugel G, Steiger K, Schlitter AM, Esposito I, Mayerle J, 2019 Loss of TLR3 and its downstream signaling accelerates acinar cell damage in the acute phase of pancreatitis. Pancreatology 19 149157. (https://doi.org/10.1016/j.pan.2018.12.005)

    • Search Google Scholar
    • Export Citation
  • Rifkin IR, Leadbetter EA, Busconi L, Viglianti G & Marshak-Rothstein A 2005 Toll-like receptors, endogenous ligands, and systemic autoimmune disease. Immunological Reviews 204 2742. (https://doi.org/10.1111/j.0105-2896.2005.00239.x)

    • Search Google Scholar
    • Export Citation
  • Segerstolpe Å, Palasantza A, Eliasson P, Andersson EM, Andreasson AC, Sun X, Picelli S, Sabirsh A, Clausen M, Bjursell MK, 2016 Single-cell transcriptome profiling of human pancreatic islets in health and type 2 diabetes. Cell Metabolism 24 593607. (https://doi.org/10.1016/j.cmet.2016.08.020)

    • Search Google Scholar
    • Export Citation
  • Shi H, Kokoeva MV, Inouye K, Tzameli I, Yin H & Flier JS 2006 TLR4 links innate immunity and fatty acid-induced insulin resistance. Journal of Clinical Investigation 116 30153025. (https://doi.org/10.1172/JCI28898)

    • Search Google Scholar
    • Export Citation
  • Stridh L, Smith PL, Naylor AS, Wang X & Mallard C 2011 Regulation of toll-like receptor 1 and -2 in neonatal mice brains after hypoxia-ischemia. Journal of Neuroinflammation 8 45. (https://doi.org/10.1186/1742-2094-8-45)

    • Search Google Scholar
    • Export Citation
  • Strodthoff D, Ma Z, Wirström T, Strawbridge RJ, Ketelhuth DF, Engel D, Clarke R, Falkmer S, Hamsten A, Hansson GK, 2015 Toll-like receptor 3 influences glucose homeostasis and beta-cell insulin secretion. Diabetes 64 34253438. (https://doi.org/10.2337/db14-0838)

    • Search Google Scholar
    • Export Citation
  • Stuart T, Butler A, Hoffman P, Hafemeister C, Papalexi E, Mauck WM 3rd, Hao Y, Stoeckius M, Smibert P & Satija R 2019 Comprehensive integration of single-cell data. Cell 177 1888.e211902.e21. (https://doi.org/10.1016/j.cell.2019.05.031)

    • Search Google Scholar
    • Export Citation
  • Sun S, Ji Y, Kersten S & Qi L 2012 Mechanisms of inflammatory responses in obese adipose tissue. Annual Review of Nutrition 32 261286. (https://doi.org/10.1146/annurev-nutr-071811-150623)

    • Search Google Scholar
    • Export Citation
  • Uchida M, Oyanagi E, Kawanishi N, Iemitsu M, Miyachi M, Kremenik MJ, Onodera S & Yano H 2014 Exhaustive exercise increases the TNF-alpha production in response to flagellin via the upregulation of toll-like receptor 5 in the large intestine in mice. Immunology Letters 158 151158. (https://doi.org/10.1016/j.imlet.2013.12.021)

    • Search Google Scholar
    • Export Citation
  • Uthaya Kumar DB, Chen CL, Liu JC, Feldman DE, Sher LS, French S, DiNorcia J, French SW, Naini BV, Junrungsee S, 2016 TLR4 signaling via NANOG cooperates with STAT3 to activate Twist1 and promote formation of tumor-initiating stem-like cells in livers of mice. Gastroenterology 150 707719. (https://doi.org/10.1053/j.gastro.2015.11.002)

    • Search Google Scholar
    • Export Citation
  • Vila IK, Badin PM, Marques MA, Monbrun L, Lefort C, Mir L, Louche K, Bourlier V, Roussel B, Gui P, 2014 Immune cell toll-like receptor 4 mediates the development of obesity- and endotoxemia-associated adipose tissue fibrosis. Cell Reports 7 11161129. (https://doi.org/10.1016/j.celrep.2014.03.062)

    • Search Google Scholar
    • Export Citation
  • Vives-Pi M, Somoza N, Fernandez-Alvarez J, Vargas F, Caro P, Alba A, Gomis R, Labeta MO & Pujol-Borrell R 2003 Evidence of expression of endotoxin receptors CD14, toll-like receptors TLR4 and TLR2 and associated molecule MD-2 and of sensitivity to endotoxin (LPS) in islet beta cells. Clinical and Experimental Immunology 133 208218. (https://doi.org/10.1046/j.1365-2249.2003.02211.x)

    • Search Google Scholar
    • Export Citation
  • Wali JA, Gurzov EN, Fynch S, Elkerbout L, Kay TW, Masters SL & Thomas HE 2014 Activation of the NLRP3 inflammasome complex is not required for stress-induced death of pancreatic islets. PLoS ONE 9 e113128. (https://doi.org/10.1371/journal.pone.0113128)

    • Search Google Scholar
    • Export Citation
  • Wen L, Ley RE, Volchkov PY, Stranges PB, Avanesyan L, Stonebraker AC, Hu C, Wong FS, Szot GL, Bluestone JA, 2008 Innate immunity and intestinal microbiota in the development of type 1 diabetes. Nature 455 11091113. (https://doi.org/10.1038/nature07336)

    • Search Google Scholar
    • Export Citation
  • Wen H, Gris D, Lei Y, Jha S, Zhang L, Huang MT, Brickey WJ & Ting JP 2011 Fatty acid-induced NLRP3-ASC inflammasome activation interferes with insulin signaling. Nature Immunology 12 408415. (https://doi.org/10.1038/ni.2022)

    • Search Google Scholar
    • Export Citation
  • Wen L, Peng J, Li Z & Wong FS 2004 The effect of innate immunity on autoimmune diabetes and the expression of toll-like receptors on pancreatic islets. Journal of Immunology 172 31733180. (https://doi.org/10.4049/jimmunol.172.5.3173)

    • Search Google Scholar
    • Export Citation
  • Westwell-Roper C, Nackiewicz D, Dan M & Ehses JA 2014 Toll-like receptors and NLRP3 as central regulators of pancreatic islet inflammation in type 2 diabetes. Immunology and Cell Biology 92 314323. (https://doi.org/10.1038/icb.2014.4)

    • Search Google Scholar
    • Export Citation
  • Ying W, Lee YS, Dong Y, Seidman JS, Yang M, Isaac R, Seo JB, Yang BH, Wollam J, Riopel M, 2019 Expansion of islet-resident macrophages leads to inflammation affecting beta cell proliferation and function in obesity. Cell Metabolism 29 457.e5474.e5. (https://doi.org/10.1016/J.cmet.2018.12.003)

    • Search Google Scholar
    • Export Citation