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BMC Nephrology

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Open Access 01-12-2015 | Research article

Pathogenic pathways are activated in each major cell type of the glomerulus in the Cd2ap mutant mouse model of focal segmental glomerulosclerosis

Authors: Eric W. Brunskill, S. Steven Potter

Published in: BMC Nephrology | Issue 1/2015

Abstract

Background

Mutations in several genes expressed in podocytes, including Cd2ap, have been associated with focal segmental glomerulosclerosis in humans. Mutant mouse models provide an opportunity to better understand the molecular pathology that drives these diseases.

Methods

In this report we use a battery of transgenic-GFP mice to facilitate the purification of all three major cell types of the glomerulus from Cd2ap mutant mice. Both microarrays and RNA-seq were used to characterize the gene expression profiles of the podocytes, mesangial cells and endothelial cells, providing a global dual platform cross-validating dataset.

Results

The mesangial cells showed increased expression of profibrotic factors, including thrombospondin, Tgfb2 and Tgfb3, as well as the angiogenesis factor Vegf. They also showed upregulation of protective genes, including Aldh1a2, involved in retinoic acid synthesis and Decorin, a Tgfb antagonist. Of interest, the mesangial cells also showed significant expression of Wt1, which has generally been considered podocyte specific. The Cd2ap mutant podocytes showed upregulation of proteases as well as genes involved in muscle and vasculature development and showed a very strong gene expression signature indicating programmed cell death. Endothelial cells showed increased expression of the leukocyte adhesion associated factors Vcam1 and Sele, as well as Midkine (promoting angiogenesis), endothelin and many genes responsive to cytokines and interferons.

Conclusions

This study provides a comprehensive analysis of the changing properties of the three cell types of the glomerulus in Cd2ap mutants, identifying activated and repressed pathways and responsible genes, thereby delivering a deeper molecular understanding of this genetic disease.

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Shih NY, Li J, Cotran R, Mundel P, Miner JH, Shaw AS. CD2AP localizes to the slit diaphragm and binds to nephrin via a novel C-terminal domain. Am J Pathol. 2001;159(6):2303–8.CrossRefPubMedPubMedCentral

Schwarz K, Simons M, Reiser J, Saleem MA, Faul C, Kriz W, et al. Podocin, a raft-associated component of the glomerular slit diaphragm, interacts with CD2AP and nephrin. J Clin Invest. 2001;108(11):1621–9.CrossRefPubMedPubMedCentral

Gigante M, Pontrelli P, Montemurno E, Roca L, Aucella F, Penza R, et al. CD2AP mutations are associated with sporadic nephrotic syndrome and focal segmental glomerulosclerosis (FSGS). Nephrol Dial Transplant. 2009;24(6):1858–64.CrossRefPubMed

Lowik MM, Groenen PJ, Pronk I, Lilien MR, Goldschmeding R, Dijkman HB, et al. Focal segmental glomerulosclerosis in a patient homozygous for a CD2AP mutation. Kidney Int. 2007;72(10):1198–203.CrossRefPubMed

Kim JM, Wu H, Green G, Winkler CA, Kopp JB, Miner JH, et al. CD2-associated protein haploinsufficiency is linked to glomerular disease susceptibility. Science. 2003;300(5623):1298–300.CrossRefPubMed

Grunkemeyer JA, Kwoh C, Huber TB, Shaw AS. CD2-associated protein (CD2AP) expression in podocytes rescues lethality of CD2AP deficiency. J Biol Chem. 2005;280(33):29677–81.CrossRefPubMed

Galkina E, Ley K. Leukocyte recruitment and vascular injury in diabetic nephropathy. J Am Soc Nephrol. 2006;17(2):368–77.CrossRefPubMed

Zent R, Pozzi A. Angiogenesis in diabetic nephropathy. Semin Nephrol. 2007;27(2):161–71.CrossRefPubMed

Shih NY, Li J, Karpitskii V, Nguyen A, Dustin ML, Kanagawa O, et al. Congenital nephrotic syndrome in mice lacking CD2-associated protein. Science. 1999;286(5438):312–5.CrossRefPubMed

10.

Motoike T, Loughna S, Perens E, Roman BL, Liao W, Chau TC, et al. Universal GFP reporter for the study of vascular development. Genesis. 2000;28(2):75–81.CrossRefPubMed

11.

Brunskill EW, Potter SS. Gene expression programs of mouse endothelial cells in kidney development and disease. PLoS ONE. 2010;5(8), e12034.CrossRefPubMedPubMedCentral

12.

Brunskill EW, Aronow BJ, Georgas K, Rumballe B, Valerius MT, Aronow J, et al. Atlas of gene expression in the developing kidney at microanatomic resolution. Dev Cell. 2008;15(5):781–91.CrossRefPubMedPubMedCentral

13.

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):W305–11.CrossRefPubMedPubMedCentral

14.

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):W96–102.CrossRefPubMedPubMedCentral

15.

Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498–504.CrossRefPubMedPubMedCentral

16.

Farquhar MG, Palade GE. Functional evidence for the existence of a third cell type in the renal glomerulus : phagocytosis of filtration residues by a distinctive “Third” cell. J Cell Biol. 1962;13(1):55–87.CrossRefPubMedPubMedCentral

17.

Brunskill EW, Potter SS. Changes in the gene expression programs of renal mesangial cells during diabetic nephropathy. BMC Nephrol. 2012;13:70.CrossRefPubMedPubMedCentral

18.

Kulkarni O, Pawar RD, Purschke W, Eulberg D, Selve N, Buchner K, et al. Spiegelmer inhibition of CCL2/MCP-1 ameliorates lupus nephritis in MRL-(Fas)lpr mice. J Am Soc Nephrol. 2007;18(8):2350–8.CrossRefPubMed

19.

Kanai T, Yamagata T, Momoi MY. Macrophage inflammatory protein-1beta and interleukin-8 associated with idiopathic steroid-sensitive nephrotic syndrome. Pediatr Int. 2009;51(4):443–7.CrossRefPubMed

20.

Ziyadeh FN. Mediators of diabetic renal disease: the case for tgf-Beta as the major mediator. J Am Soc Nephrol. 2004;15 Suppl 1:S55–7.CrossRefPubMed

21.

Crawford SE, Stellmach V, Murphy-Ullrich JE, Ribeiro SM, Lawler J, Hynes RO, et al. Thrombospondin-1 is a major activator of TGF-beta1 in vivo. Cell. 1998;93(7):1159–70.CrossRefPubMed

22.

Peired A, Angelotti ML, Ronconi E, la Marca G, Mazzinghi B, Sisti A, et al. Proteinuria impairs podocyte regeneration by sequestering retinoic acid. J Am Soc Nephrol. 2013;24(11):1756–68.CrossRefPubMedPubMedCentral

23.

Kawai T, Masaki T, Doi S, Arakawa T, Yokoyama Y, Doi T, et al. PPAR-gamma agonist attenuates renal interstitial fibrosis and inflammation through reduction of TGF-beta. Lab Investig. 2009;89(1):47–58.CrossRefPubMed

24.

Muto S, Aiba A, Saito Y, Nakao K, Nakamura K, Tomita K, et al. Pioglitazone improves the phenotype and molecular defects of a targeted Pkd1 mutant. Hum Mol Genet. 2002;11(15):1731–42.CrossRefPubMed

25.

Yoshihara D, Kurahashi H, Morita M, Kugita M, Hiki Y, Aukema HM, et al. PPAR-gamma agonist ameliorates kidney and liver disease in an orthologous rat model of human autosomal recessive polycystic kidney disease. Am J Physiol Ren Physiol. 2011;300(2):F465–74.CrossRef

26.

Ferdous Z, Peterson SB, Tseng H, Anderson DK, Iozzo RV, Grande-Allen KJ. A role for decorin in controlling proliferation, adhesion and migration of murine embryonic fibroblasts. J Biomed Mater Res Part A. 2010;93(2):419–28.

27.

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):238–45.CrossRefPubMedPubMedCentral

28.

Williams KJ, Qiu G, Usui HK, Dunn SR, McCue P, Bottinger E, et al. Decorin deficiency enhances progressive nephropathy in diabetic mice. Am J Pathol. 2007;171(5):1441–50.CrossRefPubMedPubMedCentral

29.

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.CrossRefPubMedPubMedCentral

30.

Musante L, Candiano G, Bruschi M, Zennaro C, Carraro M, Artero M, et al. Characterization of plasma factors that alter the permeability to albumin within isolated glomeruli. Proteomics. 2002;2(2):197–205.CrossRefPubMed

31.

Wenderfer SE. Can inhibition of proteasomes or NF-kappaB help control idiopathic nephrotic syndrome? Nephrol Dial Transplant. 2012;27(5):1698–701.CrossRefPubMed

32.

Schell C, Huber TB. New players in the pathogenesis of focal segmental glomerulosclerosis. Nephrol Dial Transplant. 2012;27(9):3406–12.CrossRefPubMed

33.

Sharma M, Sharma R, McCarthy ET, Savin VJ. “The FSGS factor”: enrichment and in vivo effect of activity from focal segmental glomerulosclerosis plasma. J Am Soc Nephrol. 1999;10(3):552–61.PubMed

34.

Carraro M, Zennaro C, Artero M, Candiano G, Ghiggeri GM, Musante L, et al. The effect of proteinase inhibitors on glomerular albumin permeability induced in vitro by serum from patients with idiopathic focal segmental glomerulosclerosis. Nephrol Dial Transplant. 2004;19(8):1969–75.CrossRefPubMed

35.

Bennett MR, Czech KA, Arend LJ, Witte DP, Devarajan P, Potter SS. Laser capture microdissection-microarray analysis of focal segmental glomerulosclerosis glomeruli. Nephron Exp Nephrol. 2007;107(1):e30–40.CrossRefPubMed

36.

Yaddanapudi S, Altintas MM, Kistler AD, Fernandez I, Moller CC, Wei C, et al. CD2AP in mouse and human podocytes controls a proteolytic program that regulates cytoskeletal structure and cellular survival. J Clin Invest. 2011;121(10):3965–80.CrossRefPubMedPubMedCentral

37.

Pippin JW, Sparks MA, Glenn ST, Buitrago S, Coffman TM, Duffield JS, et al. Cells of renin lineage are progenitors of podocytes and parietal epithelial cells in experimental glomerular disease. Am J Pathol. 2013;183(2):542–57.CrossRefPubMedPubMedCentral

Title: Pathogenic pathways are activated in each major cell type of the glomerulus in the Cd2ap mutant mouse model of focal segmental glomerulosclerosis
Authors: Eric W. Brunskill
S. Steven Potter
Publication date: 01-12-2015
Publisher: BioMed Central
Published in: BMC Nephrology / Issue 1/2015
Electronic ISSN: 1471-2369
DOI: https://doi.org/10.1186/s12882-015-0063-z

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Pathogenic pathways are activated in each major cell type of the glomerulus in the Cd2ap mutant mouse model of focal segmental glomerulosclerosis

Abstract

Background

Methods

Results

Conclusions

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Abstract

Background

Methods

Results

Conclusions

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