Microarray and bioinformatic detection of novel and established genes expressed in experimental anti-Thy1 nephritis.
BACKGROUND
Microarray technology is a powerful tool that can probe the molecular pathogenesis of renal injury. In this present study microarray analysis was used to monitor serial changes in the renal transcriptome of a rat model of mesangial proliferative glomerulonephritis. Administration of anti-Thy1 antibody results in phases of acute mesangial injury (day 2), cell proliferation (day 5), matrix expansion (days 5 and 7), and subsequent healing (day 14).


METHODS
Using Affymetrix (RAE230A) microarrays coupled with sequential primary biologic function-focused and secondary "baited" global cluster analysis, a cohort of established and putative novel modulators of mesangial cell turnover was identified.


RESULTS
Cluster analysis of proliferative genes identified a number of gene expression profiles. The most striking pattern was increased gene expression at day 5, a cluster that included platelet-derived growth factor (PDGF), cyclins and transforming growth factor-beta (TGF-beta). The gene expression patterns identified by primary focused cluster analysis were used as bioinformatic bait and resulted in the identification of novel families of genes such as the S100 family. The expression of established and novel genes was confirmed using reverse transcription-polymerase chain reaction (RT-PCR). Next, in vivo gene expression was compared to PDGF-stimulated mesangial cells in vitro revealing similar patterns of dysregulation.


CONCLUSIONS
Transcriptomic analysis defined both known and novel molecules involved in mesangial cell proliferation in vitro and in vivo and defined a panel of molecules that are potential contributors to mesangial cell dysfunction in glomerular disease.
Sadlier
Denise M
DM
Department of Medicine and Therapeutics, Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Ireland. dsadlier.genome@mater.ie
Ouyang
Xuesong
X
McMahon
Blaithin
B
Mu
Wei
W
Ohashi
Ryuji
R
Rodgers
Karen
K
Murray
David
D
Nakagawa
Takahiko
T
Godson
Catherine
C
Doran
Peter
P
Brady
Hugh R
HR
Johnson
Rick J
RJ
D000818:Animals; D002455:Cell Division / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D016000:Cluster Analysis; D019295:Computational Biology; D004195:Disease Models, Animal; D015432:Glomerulonephritis, Membranoproliferative / Q000235:genetics*; D006801:Humans; D007518:Isoantibodies; D050527:Mesangial Cells / Q000166:cytology / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis / Q000592:standards; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
D007518:Isoantibodies; D010982:Platelet-Derived Growth Factor; C035678:anti-Thy antibody