Endothelial-monocyte activating polypeptide II disrupts alveolar epithelial type II to type I cell transdifferentiation
Tóm tắt
Distal alveolar morphogenesis is marked by differentiation of alveolar type (AT)-II to AT-I cells that give rise to the primary site of gas exchange, the alveolar/vascular interface. Endothelial-Monocyte Activating Polypeptide (EMAP) II, an endogenous protein with anti-angiogenic properties, profoundly disrupts distal lung neovascularization and alveolar formation during lung morphogenesis, and is robustly expressed in the dysplastic alveolar regions of infants with Bronchopulmonary dysplasia. Determination as to whether EMAP II has a direct or indirect affect on ATII→ATI trans-differentiation has not been explored. In a controlled nonvascular environment, an in vitro model of ATII→ATI cell trans-differentiation was utilized to demonstrate the contribution that one vascular mediator has on distal epithelial cell differentiation. Here, we show that EMAP II significantly blocked ATII→ATI cell transdifferentiation by increasing cellular apoptosis and inhibiting expression of ATI markers. Moreover, EMAP II-treated ATII cells displayed myofibroblast characteristics, including elevated cellular proliferation, increased actin cytoskeleton stress fibers and Rho-GTPase activity, and increased nuclear:cytoplasmic volume. However, EMAP II-treated cells did not express the myofibroblast markers desmin or αSMA. Our findings demonstrate that EMAP II interferes with ATII → ATI transdifferentiation resulting in a proliferating non-myofibroblast cell. These data identify the transdifferentiating alveolar cell as a possible target for EMAP II's induction of alveolar dysplasia.
Tài liệu tham khảo
Rojas M, Woods CR, Mora AL, Xu J, Brigham KL: Endotoxin-induced lung injury in mice: structural, functional, and biochemical responses. Am J Physiol Lung Cell Mol Physiol. 2005, 288 (2): L333-341.
Santos FB, Nagato LK, Boechem NM, Negri EM, Guimaraes A, Capelozzi VL, Faffe DS, Zin WA, Rocco PR: Time course of lung parenchyma remodeling in pulmonary and extrapulmonary acute lung injury. J Appl Physiol. 2006, 100 (1): 98-106. 10.1152/japplphysiol.00395.2005.
Willis BC, duBois RM, Borok Z: Epithelial origin of myofibroblasts during fibrosis in the lung. Proc Am Thorac Soc. 2006, 3 (4): 377-382. 10.1513/pats.200601-004TK.
Willis BC, Liebler JM, Luby-Phelps K, Nicholson AG, Crandall ED, du Bois RM, Borok Z: Induction of epithelial-mesenchymal transition in alveolar epithelial cells by transforming growth factor-beta1: potential role in idiopathic pulmonary fibrosis. Am J Pathol. 2005, 166 (5): 1321-1332. 10.1016/S0002-9440(10)62351-6.
Schwarz MA, Zhang F, Gebb S, Starnes V, Warburton D: EMAP II Inhibits Lung Neovascularization and Airway Epithelial Morphogenesis. Mechanisms of Development. 2000, 95: 123-132. 10.1016/S0925-4773(00)00361-0.
Quintos-Alagheband ML, White CW, Schwarz MA: Potential role for antiangiogenic proteins in the evolution of bronchopulmonary dysplasia. Antioxid Redox Signal. 2004, 6 (1): 137-145. 10.1089/152308604771978444.
Kao J, Fan YG, Haehnel I, Clauss M, Stern D: Endothelial-monocyte activating polypeptides (EMAPs): tumor derived mediators which activate the host inflammatory response. Behring Inst Mitt. 1993, 92: 92-106.
Kao J, Houck K, Fan Y, Haehnel I, Libutti SK, Kayton ML, Grikscheit T, Chabot J, Nowygrod R, Greenberg S, et al: Characterization of a novel tumor-derived cytokine. Endothelial- monocyte activating polypeptide II. J Biol Chem. 1994, 269 (40): 25106-25119.
Kao J, Ryan J, Brett G, Chen J, Shen H, Fan YG, Godman G, Familletti PC, Wang F, Pan YC, et al: Endothelial monocyte-activating polypeptide II. A novel tumor-derived polypeptide that activates host-response mechanisms. J Biol Chem. 1992, 267 (28): 20239-20247.
Schwarz MA, Kandel J, Brett G, Li J, Hayward J, Schwarz RE, Chappey O, Wautier J, Chabot J, Lo Gerfo P, et al: Endothelial-Monocyte Activating Polypeptide II, A Novel Antitumor Cytokine that Suppresses Primary and Metastatic Tumor Growth, and Induces Apoptosis in Growing Endothelial Cells. Journal of Experimental Medicine. 1999, 190 (3): 341-353. 10.1084/jem.190.3.341.
Schwarz RE, Schwarz MA: In vivo therapy of local tumor progression by targeting vascular endothelium with EMAP-II. J Surg Res. 2004, 120 (1): 64-72. 10.1016/j.jss.2003.10.005.
Schwarz M, Lee M, Zhang F, Zhao J, Jin Y, Smith S, Bhuva J, Stern D, Warburton D, Starnes V: EMAP II: a modulator of neovascularization in the developing lung. Am J Physiol. 1999, 276 (2 Pt 1): L365-375.
Zhang F, Schwarz MA: Temporo-spatial distribution of Endothelial-monocyte Activating Polypeptide II, an anti-angiogenic protein, in the mouse embryo. Developmental Dynamics. 2000, 218: 490-498. 10.1002/1097-0177(200007)218:3<490::AID-DVDY1005>3.0.CO;2-C.
Journeay WS, Janardhan KS, Singh B: Expression and function of endothelial monocyte-activating polypeptide-II in acute lung inflammation. Inflamm Res. 2007, 56 (5): 175-181. 10.1007/s00011-006-6162-3.
Clauss M, Voswinckel R, Rajashekhar G, Sigua NL, Fehrenbach H, Rush NI, Schweitzer KS, Yildirim AO, Kamocki K, Fisher AJ, et al: Lung endothelial monocyte-activating protein 2 is a mediator of cigarette smoke-induced emphysema in mice. J Clin Invest. 2011, 121 (6): 2470-2479. 10.1172/JCI43881.
Borok Z, Danto SI, Zabski SM, Crandall ED: Defined medium for primary culture de novo of adult rat alveolar epithelial cells. In Vitro Cell Dev Biol Anim. 1994, 2: 99-104.
Danto SI, Shannon JM, Borok Z, Zabski SM, Crandall ED: Reversible transdifferentiation of alveolar epithelial cells. Am J Respir Cell Mol Biol. 1995, 12 (5): 497-502.
Kao WW, Prockop DJ: Proline analogue removes fibroblasts from cultured mixed cell populations. Nature. 1977, 266 (5597): 63-64. 10.1038/266063a0.
Robinson EE, Foty RA, Corbett SA: Fibronectin matrix assembly regulates alpha5beta1-mediated cell cohesion. Mol Biol Cell. 2004, 15 (3): 973-981.
Borok Z, Hami A, Danto SI, Zabski SM, Crandall ED: Rat serum inhibits progression of alveolar epithelial cells toward the type I cell phenotype in vitro. Am J Respir Cell Mol Biol. 1995, 12 (1): 50-55.
Schwarz MA, Wan Z, Liu J, Lee MK: Epithelial-mesenchymal interactions are linked to neovascularization. Am J Respir Cell Mol Biol. 2004, 30 (6): 784-792.
Evans MJ, Cabral LJ, Stephens RJ, Freeman G: Transformation of alveolar type 2 cells to type 1 cells following exposure to NO2. Exp Mol Pathol. 1975, 22 (1): 142-150. 10.1016/0014-4800(75)90059-3.
Dietrich C, Scherwat J, Faust D, Oesch F: Subcellular localization of beta-catenin is regulated by cell density. Biochem Biophys Res Commun. 2002, 292 (1): 195-199. 10.1006/bbrc.2002.6625.
Sharma M, Henderson BR: IQ-domain GTPase-activating protein 1 regulates beta-catenin at membrane ruffles and its role in macropinocytosis of N-cadherin and adenomatous polyposis coli. J Biol Chem. 2007, 282 (11): 8545-8556. 10.1074/jbc.M610272200.
Kalluri R, Neilson EG: Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest. 2003, 112 (12): 1776-1784.
Ullmann U, Gilles C, De Rycke M, Van de Velde H, Sermon K, Liebaers I: GSK-3-specific inhibitor-supplemented hESC medium prevents the epithelial-mesenchymal transition process and the up-regulation of matrix metalloproteinases in hESCs cultured in feeder-free conditions. Mol Hum Reprod. 2008, 14 (3): 169-179. 10.1093/molehr/gan001.
Pozharskaya V, Torres-Gonzalez E, Rojas M, Gal A, Amin M, Dollard S, Roman J, Stecenko AA, Mora AL: Twist: a regulator of epithelial-mesenchymal transition in lung fibrosis. PLoS One. 2009, 4 (10): e7559-10.1371/journal.pone.0007559.
Brown KR, England KM, Goss KL, Snyder JM, Acarregui MJ: VEGF induces airway epithelial cell proliferation in human fetal lung in vitro. Am J Physiol Lung Cell Mol Physiol. 2001, 281 (4): L1001-1010.
Kasahara Y, Tuder RM, Taraseviciene-Stewart L, Le Cras TD, Abman S, Hirth PK, Waltenberger J, Voelkel NF: Inhibition of VEGF receptors causes lung cell apoptosis and emphysema. J Clin Invest. 2000, 106 (11): 1311-1319. 10.1172/JCI10259.
Zhao L, Wang K, Ferrara N, Vu TH: Vascular endothelial growth factor co-ordinates proper development of lung epithelium and vasculature. Mech Dev. 2005, 122 (7-8): 877-886. 10.1016/j.mod.2005.04.001.
Zeng X, Wert SE, Federici R, Peters KG, Whitsett JA: VEGF enhances pulmonary vasculogenesis and disrupts lung morphogenesis in vivo. Dev Dyn. 1998, 211 (3): 215-227. 10.1002/(SICI)1097-0177(199803)211:3<215::AID-AJA3>3.0.CO;2-K.
DeLisser HM, Helmke BP, Cao G, Egan PM, Taichman D, Fehrenbach M, Zaman A, Cui Z, Mohan GS, Baldwin HS, et al: Loss of PECAM-1 function impairs alveolarization. J Biol Chem. 2006, 281 (13): 8724-8731. 10.1074/jbc.M511798200.
Sugahara K, Kiyota T, Clark RA, Mason RJ: The effect of fibronectin on cytoskeleton structure and transepithelial resistance of alveolar type II cells in primary culture. Virchows Arch B Cell Pathol Incl Mol Pathol. 1993, 64 (2): 115-122.
Dahl KN, Ribeiro AJ, Lammerding J: Nuclear shape, mechanics, and mechanotransduction. Circ Res. 2008, 102 (11): 1307-1318. 10.1161/CIRCRESAHA.108.173989.
Bhowmick NA, Ghiassi M, Bakin A, Aakre M, Lundquist CA, Engel ME, Arteaga CL, Moses HL: Transforming growth factor-beta1 mediates epithelial to mesenchymal transdifferentiation through a RhoA-dependent mechanism. Mol Biol Cell. 2001, 12 (1): 27-36.
Levayer R, Lecuit T: Breaking down EMT. Nat Cell Biol. 2008, 10 (7): 757-759. 10.1038/ncb0708-757.
Zeisberg M, Neilson EG: Biomarkers for epithelial-mesenchymal transitions. J Clin Invest. 2009, 119 (6): 1429-1437. 10.1172/JCI36183.
Kalluri R, Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest. 2009, 119 (6): 1420-1428. 10.1172/JCI39104.
Liu J, Schwarz MA: Identification of protease-sensitive sites in Human Endothelial-Monocyte Activating Polypeptide II protein. Exp Cell Res. 2006, 312 (12): 2231-2237. 10.1016/j.yexcr.2006.03.024.
Schwarz RE, Konduri S, Awasthi N, Cafasso D, Schwarz MA: An antiendothelial combination therapy strategy to increase survival in experimental pancreatic cancer. Surgery. 2009, 146 (2): 241-249. 10.1016/j.surg.2009.04.015.
Schwarz RE, Awasthi N, Konduri S, Cafasso D, Schwarz MA: EMAP II-based antiangiogenic-antiendothelial in vivo combination therapy of pancreatic cancer. Ann Surg Oncol. 2010, 17 (5): 1442-1452. 10.1245/s10434-009-0879-5.
Schwarz RE, Awasthi N, Konduri S, Caldwell L, Cafasso D, Schwarz MA: Antitumor effects of EMAP II against pancreatic cancer through inhibition of fibronectin-dependent proliferation. Cancer Biol Ther. 2010, 9 (8):
Schwarz MA, Zheng H, Liu J, Corbett S, Schwarz RE: Endothelial-monocyte activating polypeptide II alters fibronectin based endothelial cell adhesion and matrix assembly via alpha5 beta1 integrin. Exp Cell Res. 2005, 311 (2): 229-239. 10.1016/j.yexcr.2005.09.008.
Marsden M, DeSimone DW: Regulation of cell polarity, radial intercalation and epiboly in Xenopus: novel roles for integrin and fibronectin. Development. 2001, 128 (18): 3635-3647.
Davidson LA, Marsden M, Keller R, Desimone DW: Integrin alpha5beta1 and fibronectin regulate polarized cell protrusions required for Xenopus convergence and extension. Curr Biol. 2006, 16 (9): 833-844. 10.1016/j.cub.2006.03.038.
Sigurdson SL, Lwebuga-Mukasa JS: Divalent cation-dependent regulation of rat alveolar epithelial cell adhesion and spreading. Exp Cell Res. 1994, 213 (1): 71-79. 10.1006/excr.1994.1174.