DNA methylation profiling in cell models of diabetic nephropathy.
We have previously identified differentially expressed genes in cell models of diabetic nephropathy and renal biopsies. Here we have performed quantitative DNA methylation profiling in cell models of diabetic nephropathy. Over 3,000 CpG units in the promoter regions of 192 candidate genes were assessed in unstimulated human mesangial cells (HMCs) and proximal tubular epithelial cells (PTCs) compared to HMCs or PTCs exposed to appropriate stimuli. A total of 301 CpG units across 38 genes (~20%) were identified as differentially methylated in unstimulated HMCs versus PTCs. Analysis of amplicon methylation values in unstimulated versus stimulated cell models failed to demonstrate a >20% difference between amplicons. In conclusion, our results demonstrate that (1) specific DNA methylation signatures are present in HMCs and PTCs, and (2) standard protocols for exposure of renal cells to stimuli that alter gene expression may be insufficient to replicate possible alterations in DNA methylation profiles in diabetic nephropathy.
Brennan
Eoin P
EP
Nephrology Research Group, Queen’s University Belfast, UK.
Ehrich
Mathias
M
O'Donovan
Helen
H
Brazil
Derek P
DP
Crean
John K
JK
Murphy
Madeline
M
Sadlier
Denise M
DM
Martin
Finian
F
Godson
Catherine
C
van den Boom
Dirk
D
Maxwell
Alexander P
AP
Savage
David A
DA
D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D003928:Diabetic Nephropathies / Q000235:genetics*; D044127:Epigenesis, Genetic; D004847:Epithelial Cells / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D050527:Mesangial Cells / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic