Original Paper

Laser Capture Microdissection-Microarray Analysis of Focal Segmental Glomerulosclerosis Glomeruli
Michael R. Bennett^a, Kimberly A. Czech^a, Lois J. Arend^d, David P. Witte^b, Prasad Devarajan^a, S. Steven Potter^c

Divisions of
^aNephrology and Hypertension,
^bPathology and
^cDevelopmental Biology, Children's Hospital Medical Center, and
^dDepartment of Pathology and Laboratory Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, USA

Address of Corresponding Author

Nephron Exp Nephrol 2007;107:e30-e40 (DOI: 10.1159/000106775)

  Key Words

• Focal segmental glomerulosclerosis
• Laser capture
• Microarray
• Glomerulosclerosis

  Abstract

Focal segmental glomerulosclerosis (FSGS) is a major cause of end-stage renal disease. In this report we used laser capture microdissection to purify diseased glomeruli, and microarrays to provide universal gene expression profiles. The results provide a deeper understanding of the molecular mechanisms of the disease process and suggest novel therapeutic strategies. Consistent with earlier studies, molecular markers of the differentiated podocyte, including WT1, nephrin, and VEGF, were dramatically downregulated in the diseased glomerulus. We also observed multiple changes consistent with increased TGF- signaling, including elevated expression of TGF-₂, TGF-₃, SMAD2, TGF-₁ receptor, and thrombospondin. In addition, there was relatively low level expression of Csf1r, a marker of macrophages, but elevated expression of the chemokines CXCL1, CXCL2, CCL3, and CXCL14. We also observed strongly upregulated expression of Sox9, a transcription factor that can drive a genetic program of chondrogenesis and fibrosis. Further, the gene with the greatest fold increase in expression in the diseased glomerulus was osteopontin, which has been previously strongly implicated in kidney fibrosis in the unilateral ureteral obstruction mouse model. These results confirm old findings, and indicate the involvement of new genetic pathways in the cause and progression of FSGS.

Copyright © 2007 S. Karger AG, Basel

  Author Contacts

S. Steven Potter
TCHRF 3007, The Children's Hospital Medical Center
3333 Burnet Ave., Cincinnati, OH 45229 (USA)
Tel. +1 513 636 4850, Fax +1 513 636 4317
E-Mail Steve.Potter@cchmc.org

  Article Information

Supported by NIH grant DK61916 (S.S.P.)

Received: January 31, 2007
Accepted: April 24, 2007
Published online: August 6, 2007
Number of Print Pages : 11
Number of Figures : 7, Number of Tables : 1, Number of References : 46