Changes in the gene expression programs of renal mesangial cells during diabetic nephropathy

BMC Nephrology

S Steven Potter¹  Eric W Brunskill¹ 

[1] Division of Developmental Biology, Children’s Hospital Medical Center, Cincinnati, Ohio, 45229, USA

关键词: Fibrosis;    Diabetic nephropathy;    Mesangial cells;   

Others  :  1083143
DOI  :  10.1186/1471-2369-13-70

 received in 2012-05-27, accepted in 2012-07-11,  发布年份 2012

【 摘 要 】

Background

Diabetic nephropathy is the leading cause of end stage renal disease. All three cell types of the glomerulus, podocytes, endothelial cells and mesangial cells, play important roles in diabetic nephropathy. In this report we used Meis1-GFP transgenic mice to purify mesangial cells from normal mice and from db/db mice, which suffer diabetic nephropathy. The purpose of the study is to better define the unique character of normal mesangial cells, and to characterize their pathogenic and protective responses during diabetic nephropathy.

Methods

Comprehensive gene expression states of the normal and diseased mesangial cells were defined with microarrays. By comparing the gene expression profiles of mesangial cells with those of multiple other renal cell types, including podocytes, endothelial cells and renal vesicles, it was possible to better define their exceptional nature, which includes smooth muscle, phagocytic and neuronal traits.

Results

The complete set of mesangial cell expressed transcription factors, growth factors and receptors were identified. In addition, the analysis of the mesangial cells from diabetic nephropathy mice characterized their changes in gene expression. Molecular functions and biological processes specific to diseased mesangial cells were characterized, identifying genes involved in extracellular matrix, cell division, vasculogenesis, and growth factor modulation. Selected gene changes considered of particular importance to the disease process were validated and localized within the glomuerulus by immunostaining. For example, thrombospondin, a key mediator of TGFβ signaling, was upregulated in the diabetic nephropathy mesangial cells, likely contributing to fibrosis. On the other hand the decorin gene was also upregulated, and expression of this gene has been strongly implicated in the reduction of TGFβ induced fibrosis.

Conclusions

The results provide an important complement to previous studies examining mesangial cells grown in culture. The remarkable qualities of the mesangial cell are more fully defined in both the normal and diabetic nephropathy diseased state. New gene expression changes and biological pathways are discovered, yielding a deeper understanding of the diabetic nephropathy pathogenic process, and identifying candidate targets for the development of novel therapies.

【 授权许可】

   
2012 Brunskill and Potter; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Atkins RC, Zimmet P: Diabetic kidney disease: act now or pay later. Nephrol Dial Transplant 2010, 25(2):331-333.
• [2]Steffes MW, Bilous RW, Sutherland DE, Mauer SM: Cell and matrix components of the glomerular mesangium in type I diabetes. Diabetes 1992, 41(6):679-684.
• [3]Kobayashi T, Uehara S, Ikeda T, Itadani H, Kotani H: Vitamin D3 up-regulated protein-1 regulates collagen expression in mesangial cells. Kidney Int 2003, 64(5):1632-1642.
• [4]Hishikawa K, Oemar BS, Nakaki T: Static pressure regulates connective tissue growth factor expression in human mesangial cells. J Biol Chem 2001, 276(20):16797-16803.
• [5]Mishra R, Leahy P, Simonson MS: Gene expression profiling reveals role for EGF-family ligands in mesangial cell proliferation. Am J Physiol Renal Physiol 2002, 283(5):F1151-F1159.
• [6]Yoshimura H, Sakai T, Kuwahara Y, Ito M, Tsuritani K, Hirasawa Y, Nagamatsu T: Effects of kynurenine metabolites on mesangial cell proliferation and gene expression. Exp Mol Pathol 2009, 87(1):70-75.
• [7]Simonson MS, Ismail-Beigi F: Endothelin-1 increases collagen accumulation in renal mesangial cells by stimulating a chemokine and cytokine autocrine signaling loop. J Biol Chem 2011, 286(13):11003-11008.
• [8]Sadlier DM, Ouyang X, McMahon B, Mu W, Ohashi R, Rodgers K, Murray D, Nakagawa T, Godson C, Doran P, et al.: Microarray and bioinformatic detection of novel and established genes expressed in experimental anti-Thy1 nephritis. Kidney Int 2005, 68(6):2542-2561.
• [9]Brunskill EW, Georgas K, Rumballe B, Little MH, Potter SS: Defining the molecular character of the developing and adult kidney podocyte. PLoS One 2011, 6(9):e24640.
• [10]Brunskill EW, Aronow BJ, Georgas K, Rumballe B, Valerius MT, Aronow J, Kaimal V, Jegga AG, Yu J, Grimmond S, et al.: Atlas of gene expression in the developing kidney at microanatomic resolution. Dev Cell 2008, 15(5):781-791.
• [11]Chen J, Bardes EE, Aronow BJ, Jegga AG: ToppGene Suite for gene list enrichment analysis and candidate gene prioritization. Nucleic Acids Res 2009, 37(Web Server issue):W305-W311.
• [12]Kaimal V, Bardes EE, Tabar SC, Jegga AG, Aronow BJ: ToppCluster: a multiple gene list feature analyzer for comparative enrichment clustering and network-based dissection of biological systems. Nucleic Acids Res 2010, 38(Web Server issue):W96-W102.
• [13]Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, Amin N, Schwikowski B, Ideker T: Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res 2003, 13(11):2498-2504.
• [14]Hernandez AM, Villamar M, Rosello L, Moreno-Pelayo MA, Moreno F, Del Castillo I: Novel mutation in the gene encoding the GATA3 transcription factor in a Spanish familial case of hypoparathyroidism, deafness, and renal dysplasia (HDR) syndrome with female genital tract malformations. Am J Med Genet A 2007, 143(7):757-762.
• [15]Wieser R: The oncogene and developmental regulator EVI1: expression, biochemical properties, and biological functions. Gene 2007, 396(2):346-357.
• [16]Gurumurthy S, Rangnekar VM: Par-4 inducible apoptosis in prostate cancer cells. J Cell Biochem 2004, 91(3):504-512.
• [17]Xie J, Guo Q: PAR-4 is involved in regulation of beta-secretase cleavage of the Alzheimer amyloid precursor protein. J Biol Chem 2005, 280(14):13824-13832.
• [18]Johnstone RW, See RH, Sells SF, Wang J, Muthukkumar S, Englert C, Haber DA, Licht JD, Sugrue SP, Roberts T, et al.: A novel repressor, par-4, modulates transcription and growth suppression functions of the Wilms' tumor suppressor WT1. Mol Cell Biol 1996, 16(12):6945-6956.
• [19]Guenette SY, Chen J, Jondro PD, Tanzi RE: Association of a novel human FE65-like protein with the cytoplasmic domain of the beta-amyloid precursor protein. Proc Natl Acad Sci U S A 1996, 93(20):10832-10837.
• [20]Lange A, Thon L, Mathieu S, Adam D: The apoptosis inhibitory domain of FE65-like protein 1 regulates both apoptotic and caspase-independent programmed cell death mediated by tumor necrosis factor. Biochem Biophys Res Commun 2005, 335(2):575-583.
• [21]Watanabe M, Layne MD, Hsieh CM, Maemura K, Gray S, Lee ME, Jain MK: Regulation of smooth muscle cell differentiation by AT-rich interaction domain transcription factors Mrf2alpha and Mrf2beta. Circ Res 2002, 91(5):382-389.
• [22]Schmahl J, Raymond CS, Soriano P: PDGF signaling specificity is mediated through multiple immediate early genes. Nat Genet 2007, 39(1):52-60.
• [23]Chen F, Kook H, Milewski R, Gitler AD, Lu MM, Li J, Nazarian R, Schnepp R, Jen K, Biben C, et al.: Hop is an unusual homeobox gene that modulates cardiac development. Cell 2002, 110(6):713-723.
• [24]Wang B, Herman-Edelstein M, Koh P, Burns W, Jandeleit-Dahm K, Watson A, Saleem M, Goodall GJ, Twigg SM, Cooper ME, et al.: E-cadherin expression is regulated by miR-192/215 by a mechanism that is independent of the profibrotic effects of transforming growth factor-beta. Diabetes 2010, 59(7):1794-1802.
• [25]Leong KG, Niessen K, Kulic I, Raouf A, Eaves C, Pollet I, Karsan A: Jagged1-mediated Notch activation induces epithelial-to-mesenchymal transition through Slug-induced repression of E-cadherin. J Exp Med 2007, 204(12):2935-2948.
• [26]Russell L, Garrett-Sinha LA: Transcription factor Ets-1 in cytokine and chemokine gene regulation. Cytokine 2010, 51(3):217-226.
• [27]Orr B, Vanpoucke G, Grace OC, Smith L, Anderson RA, Riddick AC, Franco OE, Hayward SW, Thomson AA: Expression of pleiotrophin in the prostate is androgen regulated and it functions as an autocrine regulator of mesenchyme and cancer associated fibroblasts and as a paracrine regulator of epithelia. Prostate 2011, 71(3):305-317.
• [28]Perez-Pinera P, Alcantara S, Dimitrov T, Vega JA, Deuel TF: Pleiotrophin disrupts calcium-dependent homophilic cell-cell adhesion and initiates an epithelial-mesenchymal transition. Proc Natl Acad Sci U S A 2006, 103(47):17795-17800.
• [29]Svensson SL, Pasupuleti M, Walse B, Malmsten M, Morgelin M, Sjogren C, Olin AI, Collin M, Schmidtchen A, Palmer R, et al.: Midkine and pleiotrophin have bactericidal properties: preserved antibacterial activity in a family of heparin-binding growth factors during evolution. J Biol Chem 2010, 285(21):16105-16115.
• [30]Raymond A, Ensslin MA, Shur BD: SED1/MFG-E8: a bi-motif protein that orchestrates diverse cellular interactions. J Cell Biochem 2009, 106(6):957-966.
• [31]Motegi S, Leitner WW, Lu M, Tada Y, Sardy M, Wu C, Chavakis T, Udey MC: Pericyte-derived MFG-E8 regulates pathologic angiogenesis. Arterioscler Thromb Vasc Biol 2011, 31(9):2024-2034.
• [32]Motegi S, Garfield S, Feng X, Sardy M, Udey MC: Potentiation of platelet-derived growth factor receptor-beta signaling mediated by integrin-associated MFG-E8. Arterioscler Thromb Vasc Biol 2011, 31(11):2653-2664.
• [33]Oliverio MI, Madsen K, Best CF, Ito M, Maeda N, Smithies O, Coffman TM: Renal growth and development in mice lacking AT1A receptors for angiotensin II. Am J Physiol 1998, 274(1 Pt 2):F43-F50.
• [34]Soriano P: Abnormal kidney development and hematological disorders in PDGF beta-receptor mutant mice. Genes Dev 1994, 8(16):1888-1896.
• [35]Keul P, Lucke S, von Wnuck Lipinski K, Bode C, Graler M, Heusch G, Levkau B: Sphingosine-1-phosphate receptor 3 promotes recruitment of monocyte/macrophages in inflammation and atherosclerosis. Circ Res 2011, 108(3):314-323.
• [36]Takuwa N, Ohkura S, Takashima S, Ohtani K, Okamoto Y, Tanaka T, Hirano K, Usui S, Wang F, Du W, et al.: S1P3-mediated cardiac fibrosis in sphingosine kinase 1 transgenic mice involves reactive oxygen species. Cardiovasc Res 2010, 85(3):484-493.
• [37]Brown HJ, Lock HR, Wolfs TG, Buurman WA, Sacks SH, Robson MG: Toll-like receptor 4 ligation on intrinsic renal cells contributes to the induction of antibody-mediated glomerulonephritis via CXCL1 and CXCL2. J Am Soc Nephrol 2007, 18(6):1732-1739.
• [38]Sharma K, McCue P, Dunn SR: Diabetic kidney disease in the db/db mouse. Am J Physiol Renal Physiol 2003, 284(6):F1138-F1144.
• [39]Lawler J, Sunday M, Thibert V, Duquette M, George EL, Rayburn H, Hynes RO: Thrombospondin-1 is required for normal murine pulmonary homeostasis and its absence causes pneumonia. J Clin Invest 1998, 101(5):982-992.
• [40]Crawford SE, Stellmach V, Murphy-Ullrich JE, Ribeiro SM, Lawler J, Hynes RO, Boivin GP, Bouck N: Thrombospondin-1 is a major activator of TGF-beta1 in vivo. Cell 1998, 93(7):1159-1170.
• [41]Ziyadeh FN: Mediators of diabetic renal disease: the case for tgf-Beta as the major mediator. J Am Soc Nephrol 2004, 15(Suppl 1):S55-S57.
• [42]Raugi GJ, Lovett DH: Thrombospondin secretion by cultured human glomerular mesangial cells. Am J Pathol 1987, 129(2):364-372.
• [43]Wahab NA, Schaefer L, Weston BS, Yiannikouris O, Wright A, Babelova A, Schaefer R, Mason RM: Glomerular expression of thrombospondin-1, transforming growth factor beta and connective tissue growth factor at different stages of diabetic nephropathy and their interdependent roles in mesangial response to diabetic stimuli. Diabetologia 2005, 48(12):2650-2660.
• [44]Adler SG, Feld S, Striker L, Striker G, LaPage J, Esposito C, Aboulhosn J, Barba L, Cha DR, Nast CC: Glomerular type IV collagen in patients with diabetic nephropathy with and without additional glomerular disease. Kidney Int 2000, 57(5):2084-2092.
• [45]Leehey DJ, Song RH, Alavi N, Singh AK: Decreased degradative enzymes in mesangial cells cultured in high glucose media. Diabetes 1995, 44(8):929-935.
• [46]Lam S, van der Geest RN, Verhagen NA, Daha MR, van Kooten C: Secretion of collagen type IV by human renal fibroblasts is increased by high glucose via a TGF-beta-independent pathway. Nephrol Dial Transplant 2004, 19(7):1694-1701.
• [47]Whiteside CI, Dlugosz JA: Mesangial cell protein kinase C isozyme activation in the diabetic milieu. Am J Physiol Renal Physiol 2002, 282(6):F975-F980.
• [48]Kinoshita M, Nakamura T, Ihara M, Haraguchi T, Hiraoka Y, Tashiro K, Noda M: Identification of human endomucin-1 and −2 as membrane-bound O-sialoglycoproteins with anti-adhesive activity. FEBS Lett 2001, 499(1–2):121-126.
• [49]Larsson M, Majeed M, Ernst JD, Magnusson KE, Stendahl O, Forsum U: Role of annexins in endocytosis of antigens in immature human dendritic cells. Immunology 1997, 92(4):501-511.
• [50]Park JE, Lee DH, Lee JA, Park SG, Kim NS, Park BC, Cho S: Annexin A3 is a potential angiogenic mediator. Biochem Biophys Res Commun 2005, 337(4):1283-1287.
• [51]Pu L, Fan JM: Effects of pioglitazone on the transdifferentiation and CTGF expression of renal tubular epithelial-myofibroblast in vitro. Sichuan Da Xue Xue Bao Yi Xue Ban 2010, 41(1):15-19.
• [52]Ferdous Z, Wei VM, Iozzo R, Hook M, Grande-Allen KJ: Decorin-transforming growth factor- interaction regulates matrix organization and mechanical characteristics of three-dimensional collagen matrices. J Biol Chem 2007, 282(49):35887-35898.
• [53]Mohan RR, Gupta R, Mehan MK, Cowden JW, Sinha S: Decorin transfection suppresses profibrogenic genes and myofibroblast formation in human corneal fibroblasts. Exp Eye Res 2010, 91(2):238-245.
• [54]Williams KJ, Qiu G, Usui HK, Dunn SR, McCue P, Bottinger E, Iozzo RV, Sharma K: Decorin deficiency enhances progressive nephropathy in diabetic mice. Am J Pathol 2007, 171(5):1441-1450.