Abstract
The receptor for advanced glycation endproducts (RAGE) is expressed under pathological conditions in many tissues and has been assigned many functions. We demonstrate, in normal human lung tissue, the preferential and highly abundant expression of RAGE by quantitative polymerase chain reaction. In addition, RAGE expression, as a specific differentiation marker of alveolar epithelial type I cells (AT I cells), and its localization to the basolateral plasma membrane have been confirmed by means of newly raised monoclonal antibodies. The physiological function of RAGE on AT I cells has previously remained elusive. By using HEK293 cells transfected with cDNA encoding for full-length RAGE, we show that RAGE enhances the adherence of epithelial cells to collagen-coated surfaces and has a striking capacity for inducing cell spreading. The preferential binding of RAGE to collagen has been confirmed by assaying the binding of soluble RAGE to various substrates. RAGE might thus assist AT I cells to acquire a spreading morphology, thereby ensuring effective gas exchange and alveolar stability.
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Abbreviations
- AGE:
-
advanced glycation endproducts
- AT I/II:
-
alveolar epithelial type I/II cells
- CML:
-
carboxymethyllysine
- FI:
-
fluorescence intensity
- hAEpC:
-
human alveolar epithelial cells
- HUVEC:
-
human umbilical vein endothelial cells
- KGF:
-
keratinocyte growth factor, -/- mouse: knockout mouse
- PI:
-
propidium iodide
- SP-C/D:
-
surfactant protein C/D
- sRAGE:
-
soluble RAGE
- RAGE:
-
receptor for AGE
- TEER:
-
transepithelial electrical resistance
References
Bankston PW, Porter GA, Milici AJ, Palade GE (1991) Differential and specific labeling of epithelial and vascular endothelial cells of the rat lung by Lycopersicon esculentum and Griffonia simplicifolia I lectins. Eur J Cell Biol 54:187–195
Bartling B, Hofmann HS, Weigle B, Silber RE, Simm A (2005) Down-regulation of the receptor for advanced glycation end-products (RAGE) supports non-small cell lung carcinoma. Carcinogenesis 26:293–301
Bierhaus A, Illmer T, Kasper M, Luther T, Quehenberger P, Tritschler H, Wahl P, Ziegler R, Muller M, Nawroth PP (1997) Advanced glycation end product (AGE)-mediated induction of tissue factor in cultured endothelial cells is dependent on RAGE. Circulation 96:2262–2271
Bierhaus A, Schiekofer S, Schwaninger M, Andrassy M, Humpert PM, Chen J, Hong M, Luther T, Henle T, Kloting I, Morcos M, Hofmann M, Tritschler H, Weigle B, Kasper M, Smith M, Perry G, Schmidt AM, Stern DM, Haring HU, Schleicher E, Nawroth PP (2001) Diabetes-associated sustained activation of the transcription factor nuclear factor-kappaB. Diabetes 50:2792–2808
Borok Z, Lubman RL, Danto SI, Zhang XL, Zabski SM, King LS, Lee DM, Agre P, Crandall ED (1998) Keratinocyte growth factor modulates alveolar epithelial cell phenotype in vitro: expression of aquaporin 5. Am J Respir Cell Mol Biol 18:554–561
Boulanger E, Wautier MP, Wautier JL, Boval B, Panis Y, Wernert N, Danze PM, Dequiedt P (2002) AGEs bind to mesothelial cells via RAGE and stimulate VCAM-1 expression. Kidney Int 61:148–156
Brett J, Schmidt AM, Yan SD, Zou YS, Weidman E, Pinsky D, Nowygrod R, Neeper M, Przysiecki C, Shaw A (1993) Survey of the distribution of a newly characterized receptor for advanced glycation end products in tissues. Am J Pathol 143:1699–1712
Chavakis T, Bierhaus A, Al Fakhri N, Schneider D, Witte S, Linn T, Nagashima M, Morser J, Arnold B, Preissner KT, Nawroth PP (2003) The pattern recognition receptor (RAGE) is a counterreceptor for leukocyte integrins: a novel pathway for inflammatory cell recruitment. J Exp Med 198:1507–1515
Chen Z, Jin N, Narasaraju T, Chen J, McFarland LR, Scott M, Liu L (2004) Identification of two novel markers for alveolar epithelial type I and II cells. Biochem Biophys Res Commun 319:774–780
Crapo JD, Barry BE, Gehr P, Bachofen M, Weibel ER (1982) Cell number and cell characteristics of the normal human lung. Am Rev Respir Dis 126:332–337
Dahlin K, Mager EM, Allen L, Tigue Z, Goodglick L, Wadehra M, Dobbs L (2004) Identification of genes differentially expressed in rat alveolar type I cells. Am J Respir Cell Mol Biol 31:309–316
Danto SI, Shannon JM, Borok Z, Zabski SM, Crandall ED (1995) Reversible transdifferentiation of alveolar epithelial cells. Am J Respir Cell Mol Biol 12:497–502
Dobbs LG, Gonzalez R, Matthay MA, Carter EP, Allen L, Verkman AS (1998) Highly water-permeable type I alveolar epithelial cells confer high water permeability between the airspace and vasculature in rat lung. Proc Natl Acad Sci USA 95:2991–2996
Ehrhardt C, Kim KJ, Lehr CM (2004) Isolation and culture of human alveolar epithelial cells. Methods Mol Med 107:207–216
Elbert KJ, Schafer UF, Schafers HJ, Kim KJ, Lee VH, Lehr CM (1999) Monolayers of human alveolar epithelial cells in primary culture for pulmonary absorption and transport studies. Pharm Res 16:601–608
Fehrenbach H, Kasper M, Tschernig T, Shearman MS, Schuh D, Muller M (1998) Receptor for advanced glycation endproducts (RAGE) exhibits highly differential cellular and subcellular localisation in rat and human lung. Cell Mol Biol (Noisy-le-grand) 44:1147–1157
Forbes B (2002) Pulmonary epithelial cell culture. Methods Mol Biol 188:65–75
Forbes B, Shah A, Martin GP, Lansley AB (2003) The human bronchial epithelial cell line 16HBE14o- as a model system of the airways for studying drug transport. Int J Pharm 257:161–167
Fuchs S, Hollins AJ, Laue M, Schaefer UF, Roemer K, Gumbleton M, Lehr CM (2003) Differentiation of human alveolar epithelial cells in primary culture: morphological characterization and synthesis of caveolin-1 and surfactant protein-C. Cell Tissue Res 311:31–45
Geiser T (2003) Mechanisms of alveolar epithelial repair in acute lung injury–a translational approach. Swiss Med Wkly 133:586–590
Goldmann T, Vollmer E, Gerdes J (2003) What’s cooking? Detection of important biomarkers in HOPE-fixed, paraffin-embedded tissues eliminates the need for antigen retrieval. Am J Pathol 163:2638–2640
Hori O, Brett J, Slattery T, Cao R, Zhang J, Chen JX, Nagashima M, Lundh ER, Vijay S, Nitecki D (1995) The receptor for advanced glycation end products (RAGE) is a cellular binding site for amphoterin. Mediation of neurite outgrowth and co-expression of rage and amphoterin in the developing nervous system. J Biol Chem 270:25752–25761
Horvat R, Hovorka A, Dekan G, Poczewski H, Kerjaschki D (1986) Endothelial cell membranes contain podocalyxin—the major sialoprotein of visceral glomerular epithelial cells. J Cell Biol 102:484–491
Hsieh HL, Schafer BW, Weigle B, Heizmann CW (2004) S100 protein translocation in response to extracellular S100 is mediated by receptor for advanced glycation endproducts in human endothelial cells. Biochem Biophys Res Commun 316:949–959
Hudson BI, Bucciarelli LG, Wendt T, Sakaguchi T, Lalla E, Qu W, Lu Y, Lee L, Stern DM, Naka Y, Ramasamy R, Yan SD, Yan SF, D’Agati V, Schmidt AM (2003) Blockade of receptor for advanced glycation endproducts: a new target for therapeutic intervention in diabetic complications and inflammatory disorders. Arch Biochem Biophys 419:80–88
Isakson BE, Lubman RL, Seedorf GJ, Boitano S (2001) Modulation of pulmonary alveolar type II cell phenotype and communication by extracellular matrix and KGF. Am J Physiol Cell Physiol 281:C1291–C1299
Kasper M, Singh G (1995) Epithelial lung cell marker: current tools for cell typing. Histol Histopathol 10:155–169
Kasper M, Reimann T, Hempel U, Wenzel KW, Bierhaus A, Schuh D, Dimmer V, Haroske G, Muller M (1998) Loss of caveolin expression in type I pneumocytes as an indicator of subcellular alterations during lung fibrogenesis. Histochem Cell Biol 109:41–48
Kasper M, Bierhaus A, Nawroth PP (2003) Pulmonary pathology of knockouts in mice lacking type I pneumocyte antigens. Ann Anat 185(Suppl):A144
Katsuoka F, Kawakami Y, Arai T, Imuta H, Fujiwara M, Kanma H, Yamashita K (1997) Type II alveolar epithelial cells in lung express receptor for advanced glycation end products (RAGE) gene. Biochem Biophys Res Commun 238:512–516
Kim KJ, Malik AB (2003) Protein transport across the lung epithelial barrier. Am J Physiol Lung Cell Mol Physiol 284:L247–L259
Kislinger T, Fu C, Huber B, Qu W, Taguchi A, Du YS, Hofmann M, Yan SF, Pischetsrieder M, Stern D, Schmidt AM (1999) N(epsilon)-(carboxymethyl)lysine adducts of proteins are ligands for receptor for advanced glycation end products that activate cell signaling pathways and modulate gene expression. J Biol Chem 274:31740–31749
Liliensiek B, Weigand MA, Bierhaus A, Nicklas W, Kasper M, Hofer S, Plachky J, Grone HJ, Kurschus FC, Schmidt AM, Yan SD, Martin E, Schleicher E, Stern DM, Hammerling GG, Nawroth PP, Arnold B (2004) Receptor for advanced glycation end products (RAGE) regulates sepsis but not the adaptive immune response. J Clin Invest 113:1641–1650
Nagai N, Hosokawa M, Itohara S, Adachi E, Matsushita T, Hosokawa N, Nagata K (2000) Embryonic lethality of molecular chaperone hsp47 knockout mice is associated with defects in collagen biosynthesis. J Cell Biol 150:1499–1506
Neeper M, Schmidt AM, Brett J, Yan SD, Wang F, Pan YC, Elliston K, Stern D, Shaw A (1992) Cloning and expression of a cell surface receptor for advanced glycosylation end products of proteins. J Biol Chem 267:14998–15004
Phelps DS, Floros J (1991) Localization of pulmonary surfactant proteins using immunohistochemistry and tissue in situ hybridization. Exp Lung Res 17:985–995
Portnoy J, Curran-Everett D, Mason RJ (2004) Keratinocyte growth factor stimulates alveolar type II cell proliferation through the extracellular signal-regulated kinase and phosphatidylinositol 3-OH kinase pathways. Am J Respir Cell Mol Biol 30:901–907
Roth J, Taatjes DJ, Tokuyasu KT (1990) Contrasting of Lowicryl K4M thin sections. Histochemistry 95:123–136
Sakurai S, Yonekura H, Yamamoto Y, Watanabe T, Tanaka N, Li H, Rahman AK, Myint KM, Kim CH, Yamamoto H (2003) The AGE-RAGE system and diabetic nephropathy. J Am Soc Nephrol 14:S259–S263
Schmidt AM, Vianna M, Gerlach M, Brett J, Ryan J, Kao J, Esposito C, Hegarty H, Hurley W, Clauss M (1992) Isolation and characterization of two binding proteins for advanced glycosylation end products from bovine lung which are present on the endothelial cell surface. J Biol Chem 267:14987–14997
Schmidt AM, Hori O, Chen JX, Li JF, Crandall J, Zhang J, Cao R, Yan SD, Brett J, Stern D (1995) Advanced glycation endproducts interacting with their endothelial receptor induce expression of vascular cell adhesion molecule-1 (VCAM-1) in cultured human endothelial cells and in mice. A potential mechanism for the accelerated vasculopathy of diabetes. J Clin Invest 96:1395–1403
Schmidt AM, Yan SD, Yan SF, Stern DM (2001) The multiligand receptor RAGE as a progression factor amplifying immune and inflammatory responses. J Clin Invest 108:949–955
Schnittler HJ, Wilke A, Gress T, Suttorp N, Drenckhahn D (1990) Role of actin and myosin in the control of paracellular permeability in pig, rat and human vascular endothelium. J Physiol (Lond) 431:379–401
Shirasawa M, Fujiwara N, Hirabayashi S, Ohno H, Iida J, Makita K, Hata Y (2004) Receptor for advanced glycation end-products is a marker of type I lung alveolar cells. Genes Cells 9:165–174
Stern D, Yan SD, Yan SF, Schmidt AM (2002) Receptor for advanced glycation endproducts: a multiligand receptor magnifying cell stress in diverse pathologic settings. Adv Drug Deliv Rev 54:1615–1625
Taguchi A, Blood DC, Toro G del, Canet A, Lee DC, Qu W, Tanji N, Lu Y, Lalla E, Fu C, Hofmann MA, Kislinger T, Ingram M, Lu A, Tanaka H, Hori O, Ogawa S, Stern DM, Schmidt AM (2000) Blockade of RAGE-amphoterin signalling suppresses tumour growth and metastases. Nature 405:354–360
Treutiger CJ, Mullins GE, Johansson AS, Rouhiainen A, Rauvala HM, Erlandsson-Harris H, Andersson U, Yang H, Tracey KJ, Andersson J, Palmblad JE (2003) High mobility group 1 B-box mediates activation of human endothelium. J Intern Med 254:375–385
Uhal BD (1997) Cell cycle kinetics in the alveolar epithelium. Am J Physiol 272:L1031–L1045
Williams MC (2003) Alveolar type I cells: molecular phenotype and development. Annu Rev Physiol 65:669–695
Yan SD, Chen X, Fu J, Chen M, Zhu H, Roher A, Slattery T, Zhao L, Nagashima M, Morser J, Migheli A, Nawroth P, Stern D, Schmidt AM (1996) RAGE and amyloid-beta peptide neurotoxicity in Alzheimer’s disease. Nature 382:685–691
Yonekura H, Yamamoto Y, Sakurai S, Petrova RG, Abedin MJ, Li H, Yasui K, Takeuchi M, Makita Z, Takasawa S, Okamoto H, Watanabe T, Yamamoto H (2003) Novel splice variants of the receptor for advanced glycation end-products expressed in human vascular endothelial cells and pericytes, and their putative roles in diabetes-induced vascular injury. Biochem J 370:1097–1109
Acknowledgements
We thank Dr. M. A. Shearman (Merck, Sharp & Dome, Essex, UK) for kindly providing the polyclonal goat anti-RAGE serum, and Dr. S. E. Gabriel (Chapel Hill, USA) for providing the polyclonal rabbit anti-aquaporin-5 antiserum. We are also grateful to Sylvia Großklaus for isolating HUVEC, to Christine Graefe and Heidi Gebauer for immunizing mice and cultivating the hybridoma cells, and to Ms. Susanne Kossek and Ms. Katja Klein for their skilful technical assistance. Last but not least, our special thanks go to Dr. M. Bachmann (Dresden, Germany) for carefully reading the manuscript and for his inspiring discussions.
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This work was in part supported by grants from Saxonian Ministry for Science and Arts (AZ: 4-7531.50-03-0370-01/7) to N.D. and a “Förderung des wissenschaftlichen Nachwuchses” grant from the DPhG to C.E.
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Demling, N., Ehrhardt, C., Kasper, M. et al. Promotion of cell adherence and spreading: a novel function of RAGE, the highly selective differentiation marker of human alveolar epithelial type I cells. Cell Tissue Res 323, 475–488 (2006). https://doi.org/10.1007/s00441-005-0069-0
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DOI: https://doi.org/10.1007/s00441-005-0069-0