Glucose-induced gradual phenotypic modulation of cultured human glomerular epithelial cells may be independent of Wilms’ tumor 1 (WT1)
Tóm tắt
Renal podocytes form the main filtration barrier possessing a unique phenotype maintained by proteins including podocalyxin and nephrin, the expression of which is suppressed in pathological conditions. We used an in vitro model of human glomerular epithelial cells (HGEC) to investigate the role of high glucose in dysregulating the podocytic epithelial phenotype and determined the time needed for this change to occur. In our in vitro podocyte system changes indicating podocyte dedifferentiation in the prolonged presence of high glucose included loss of podocalyxin, nephrin and CD10/CALLA concomitant with upregulation of mesenchymal vimentin. Our study demonstrates for the first time that podocyte-specific markers undergo changes of expression at different time intervals, since glucose-mediated podocalyxin downregulation is a progressive process that precedes downregulation of nephrin expression. Finally we demonstrate that high glucose permanently impaired WT1 binding to the podocalyxin gene promoter region but did not affect WT1 binding on the nephrin gene promoter region. The presence of high glucose induced a phenotypic conversion of podocytes resembling partial dedifferentiation. Our study demonstrates that dysregulation of the normal podocytic phenotype is an event differentially affecting the expression of function-specific podocytic markers, exhibiting downregulation of the epithelial marker CD10/CALLA and PC first, followed by stably downregulated nephrin. Furthermore, it is herein suggested that WT1 may not be directly involved with upregulation of previously reduced PC and nephrin expression.
Tài liệu tham khảo
Reiser J, Kriz W, Kretzler M, Mundel P: The glomerular slit diaphragm is a modified adherens junction. J Am Soc Nephrol. 2000, 11 (1): 1-8.
Ronco P: Proteinuria: is it all in the foot?. J Clin Invest. 2007, 117 (8): 2079-2082. 10.1172/JCI32966.
Liu Y: New insights into epithelial-mesenchymal transition in kidney fibrosis. J Am Soc Nephrol. 2010, 21 (2): 212-222. 10.1681/ASN.2008121226.
Rhyu DY, Yang Y, Ha H, Lee GT, Song JS, Uh ST, Lee HB: Role of reactive oxygen species in TGF-beta1-induced mitogen-activated protein kinase activation and epithelial-mesenchymal transition in renal tubular epithelial cells. J Am Soc Nephrol. 2005, 16 (3): 667-675. 10.1681/ASN.2004050425.
Li Y, Kang YS, Dai C, Kiss LP, Wen X, Liu Y: Epithelial-to-mesenchymal transition is a potential pathway leading to podocyte dysfunction and proteinuria. Am J Path. 2008, 172 (2): 299-308. 10.2353/ajpath.2008.070057.
Barisoni L, Schnaper HW, Kopp JB: A proposed taxonomy for the podocytopathies: a reassessment of the primary nephrotic diseases. Clin J Am Soc Nephrol. 2007, 2 (3): 529-542. 10.2215/CJN.04121206.
Herman-Edelstein M, Thomas MC, Thallas-Bonke V, Saleem M, Cooper ME, Kantharidis P: Dedifferentiation of immortalized human podocytes in response to transforming growth factor-beta: a model for diabetic podocytopathy. Diabetes. 2011, 60 (6): 1779-1788. 10.2337/db10-1110.
Kerjaschki D, Sharkey DJ, Farquhar MG: Identification and characterization of podocalyxin–the major sialoprotein of the renal glomerular epithelial cell. J Cell Biol. 1984, 98 (4): 1591-1596. 10.1083/jcb.98.4.1591.
Kershaw DB, Beck SG, Wharram BL, Wiggins JE, Goyal M, Thomas PE, Wiggins RC: Molecular cloning and characterization of human podocalyxin-like protein. Orthologous relationship to rabbit PCLP1 and rat podocalyxin. J Biol Chem. 1997, 272 (25): 15708-15714. 10.1074/jbc.272.25.15708.
Tryggvason K: Nephrin: role in normal kidney and in disease. Adv Nephrol Necker Hosp. 2001, 31: 221-234.
Economou CG, Kitsiou PV, Tzinia AK, Panagopoulou E, Marinos E, Kershaw DB, Kerjaschki D, Tsilibary EC: Enhanced podocalyxin expression alters the structure of podocyte basal surface. J Cell Sci. 2004, 117 (Pt 15): 3281-3294.
Drossopoulou GI, Tsotakos NE, Tsilibary EC: Impaired transcription factor interplay in addition to advanced glycation end products suppress podocalyxin expression in high glucose-treated human podocytes. Am J Physiol Renal Physiol. 2009, 297 (3): F594-F603. 10.1152/ajprenal.00203.2009.
Benigni A, Gagliardini E, Tomasoni S, Abbate M, Ruggenenti P, Kalluri R, Remuzzi G: Selective impairment of gene expression and assembly of nephrin in human diabetic nephropathy. Kidney Int. 2004, 65 (6): 2193-2200. 10.1111/j.1523-1755.2004.00636.x.
Miner JH: Focusing on the glomerular slit diaphragm: podocin enters the picture. Am J Path. 2002, 160 (1): 3-5. 10.1016/S0002-9440(10)64341-6.
Zhang C, Jiang HJ, Chang Y, Fang Z, Sun XF, Liu JS, Deng AG, Zhu ZH: Downregulation of CD2-associated protein impaired the physiological functions of podocytes. Cell Biol Int. 2009, 33 (6): 632-639. 10.1016/j.cellbi.2009.02.017.
Cutrona G, Tasso P, Dono M, Roncella S, Ulivi M, Carpaneto EM, Fontana V, Comis M, Morabito F, Spinelli M: CD10 is a marker for cycling cells with propensity to apoptosis in childhood ALL. Br J Cancer. 2002, 86 (11): 1776-1785. 10.1038/sj.bjc.6600329.
Delarue F, Virone A, Hagege J, Lacave R, Peraldi MN, Adida C, Rondeau E, Feunteun J, Sraer JD: Stable cell line of T-SV40 immortalized human glomerular visceral epithelial cells. Kidney Int. 1991, 40 (5): 906-912. 10.1038/ki.1991.292.
Zeisberg M, Neilson EG: Biomarkers for epithelial-mesenchymal transitions. J Clin Inv. 2009, 119 (6): 1429-1437. 10.1172/JCI36183.
Galichon P, Hertig A: Epithelial to mesenchymal transition as a biomarker in renal fibrosis: are we ready for the bedside?. Fibrogenesis Tissue Repair. 2011, 4: 11-10.1186/1755-1536-4-11.
Carew RM, Wang B, Kantharidis P: The role of EMT in renal fibrosis. Cell Tissue Res. 2012, 347 (1): 103-116. 10.1007/s00441-011-1227-1.
Zou J, Yaoita E, Watanabe Y, Yoshida Y, Nameta M, Li H, Qu Z, Yamamoto T: Upregulation of nestin, vimentin, and desmin in rat podocytes in response to injury. Virchows Arch. 2006, 448 (4): 485-492. 10.1007/s00428-005-0134-9.
Zou J, Yaoita E, Watanabe Y, Yoshida Y, Nameta M, Li H, Qu Z, Yamamoto T: Upregulation of nestin, vimentin, and desmin in rat podocytes in response to injury. Virchows Arch. 2006, 448 (4): 485-492. 10.1007/s00428-005-0134-9.
Doublier S, Ruotsalainen V, Salvidio G, Lupia E, Biancone L, Conaldi PG, Reponen P, Tryggvason K, Camussi G: Nephrin redistribution on podocytes is a potential mechanism for proteinuria in patients with primary acquired nephrotic syndrome. Am J Path. 2001, 158 (5): 1723-1731. 10.1016/S0002-9440(10)64128-4.
Nakagama H, Heinrich G, Pelletier J, Housman DE: Sequence and structural requirements for high-affinity DNA binding by the WT1 gene product. Mol Cellular Biol. 1995, 15 (3): 1489-1498.
Palmer RE, Kotsianti A, Cadman B, Boyd T, Gerald W, Haber DA: WT1 regulates the expression of the major glomerular podocyte membrane protein Podocalyxin. Curr Biol. 2001, 11 (22): 1805-1809. 10.1016/S0960-9822(01)00560-7.
Wagner N, Wagner KD, Xing Y, Scholz H, Schedl A: The major podocyte protein nephrin is transcriptionally activated by the Wilms' tumor suppressor WT1. J Am Soc Nephrol. 2004, 15 (12): 3044-3051. 10.1097/01.ASN.0000146687.99058.25.
Ziyadeh FN, Wolf G: Pathogenesis of the podocytopathy and proteinuria in diabetic glomerulopathy. Curr Diabetes Rev. 2008, 4 (1): 39-45. 10.2174/157339908783502370.
Li JJ, Kwak SJ, Jung DS, Kim JJ, Yoo TH, Ryu DR, Han SH, Choi HY, Lee JE, Moon SJ: Podocyte biology in diabetic nephropathy. Kidney Int Suppl. 2007, 106: S36-42.
Shankland SJ: The podocyte's response to injury: role in proteinuria and glomerulosclerosis. Kidney Int. 2006, 69 (12): 2131-2147. 10.1038/sj.ki.5000410.
Fan JM, Huang XR, Ng YY, Nikolic-Paterson DJ, Mu W, Atkins RC, Lan HY: Interleukin-1 induces tubular epithelial-myofibroblast transdifferentiation through a transforming growth factor-beta1-dependent mechanism in vitro. Am J Kidney Dis. 2001, 37 (4): 820-831. 10.1016/S0272-6386(01)80132-3.
Lan HY: Tubular epithelial-myofibroblast transdifferentiation mechanisms in proximal tubule cells. Curr Opin Nephrol Hypertens. 2003, 12 (1): 25-29. 10.1097/00041552-200301000-00005.
Zhao L, Yaoita E, Nameta M, Zhang Y, Cuellar LM, Fujinaka H, Xu B, Yoshida Y, Hatakeyama K, Yamamoto T: Claudin-6 localized in tight junctions of rat podocytes. Am J Physiol Regul Integr Comp Physiol. 2008, 294 (6): R1856-1862. 10.1152/ajpregu.00862.2007.
Chen S, Lee JS, Iglesias-de la Cruz MC, Wang A, Izquierdo-Lahuerta A, Gandhi NK, Danesh FR, Wolf G, Ziyadeh FN: Angiotensin II stimulates alpha3(IV) collagen production in mouse podocytes via TGF-beta and VEGF signalling: implications for diabetic glomerulopathy. Nephrol Dial Transplant. 2005, 20 (7): 1320-1328. 10.1093/ndt/gfh837.
Wu DT, Bitzer M, Ju W, Mundel P, Bottinger EP: TGF-beta concentration specifies differential signaling profiles of growth arrest/differentiation and apoptosis in podocytes. J Am Soc Nephrol. 2005, 16 (11): 3211-3221. 10.1681/ASN.2004121055.
Susztak K, Raff AC, Schiffer M, Bottinger EP: Glucose-induced reactive oxygen species cause apoptosis of podocytes and podocyte depletion at the onset of diabetic nephropathy. Diabetes. 2006, 55 (1): 225-233. 10.2337/diabetes.55.01.06.db05-0894.
Benzing T: Signaling at the slit diaphragm. J Am Soc Nephrol. 2004, 15 (6): 1382-1391. 10.1097/01.ASN.0000130167.30769.55.
Saleem MA, O'Hare MJ, Reiser J, Coward RJ, Inward CD, Farren T, Xing CY, Ni L, Mathieson PW, Mundel P: A conditionally immortalized human podocyte cell line demonstrating nephrin and podocin expression. J Am Soc Nephrol. 2002, 13 (3): 630-638.
Meng X, Ezzati P, Wilkins JA: Requirement of podocalyxin in TGF-beta induced epithelial mesenchymal transition. PLoS One. 2011, 6 (4): e18715-10.1371/journal.pone.0018715.
Matsusaka T, Xin J, Niwa S, Kobayashi K, Akatsuka A, Hashizume H, Wang QC, Pastan I, Fogo AB, Ichikawa I: Genetic engineering of glomerular sclerosis in the mouse via control of onset and severity of podocyte-specific injury. J Am Soc Nephrol. 2005, 16 (4): 1013-1023. 10.1681/ASN.2004080720.
Guo JK, Menke AL, Gubler MC, Clarke AR, Harrison D, Hammes A, Hastie ND, Schedl A: WT1 is a key regulator of podocyte function: reduced expression levels cause crescentic glomerulonephritis and mesangial sclerosis. Hum Mol Genet. 2002, 11 (6): 651-659. 10.1093/hmg/11.6.651.
Butta N, Larrucea S, Alonso S, Rodriguez RB, Arias-Salgado EG, Ayuso MS, Gonzalez-Manchon C, Parrilla R: Role of transcription factor Sp1 and CpG methylation on the regulation of the human podocalyxin gene promoter. BMC Mol Biol. 2006, 7: 17-10.1186/1471-2199-7-17.
Beltcheva O, Hjorleifsdottir EE, Kontusaari S, Tryggvason K: Sp1 specifically binds to an evolutionarily conserved DNA segment within a region necessary for podocyte-specific expression of nephrin. Nephron Exp Nephrol. 2010, 114 (1): e15-22. 10.1159/000245062.
Deb DK, Wang Y, Zhang Z, Nie H, Huang X, Yuan Z, Chen Y, Zhao Q, Li YC: Molecular mechanism underlying 1,25-dihydroxyvitamin D regulation of nephrin gene expression. J Biol Chem. 2011, 286 (37): 32011-32017. 10.1074/jbc.M111.269118.
Kitsiou PV, Tzinia AK, Stetler-Stevenson WG, Michael AF, Fan WW, Zhou B, Tsilibary EC: Glucose-induced changes in integrins and matrix-related functions in cultured human glomerular epithelial cells. Am J Physiol Renal Physiol. 2003, 284 (4): F671-679.
Collino F, Bussolati B, Gerbaudo E, Marozio L, Pelissetto S, Benedetto C, Camussi G: Preeclamptic sera induce nephrin shedding from podocytes through endothelin-1 release by endothelial glomerular cells. Am J Physiol Renal Physiol. 2008, 294 (5): F1185-1194. 10.1152/ajprenal.00442.2007.
Chakrabarti SK, James JC, Mirmira RG: Quantitative assessment of gene targeting in vitro and in vivo by the pancreatic transcription factor, Pdx1 Importance of chromatin structure in directing promoter binding. J Biol Chem. 2002, 277 (15): 13286-13293. 10.1074/jbc.M111857200.