Comparison of different normalization strategies for the analysis of glomerular microRNAs in IgA nephropathy
Bockmeyer CL, Saeuberlich K, Wittig J, Esser M, Röder S, Vester U, Hoyer PF, Agustian PA, Zeuschner P, Amann KU, Daniel C, Becker JU (2016)
Publication Type: Journal article
Publication year: 2016
Journal
Book Volume: 6
Pages Range: 31992
DOI: 10.1038/srep31992
Abstract
Small nucleolar RNAs (snoRNAs) have been used for normalization in glomerular microRNA (miRNA) quantification without confirmation of validity. Our aim was to identify glomerular reference miRNAs in IgA nephropathy. We compared miRNAs in human paraffin-embedded renal biopsies from patients with cellular-crescentic IgA-GN (n = 5; crescentic IgA-GN) and non-crescentic IgA-GN (n = 5; IgA-GN) to mild interstitial nephritis without glomerular abnormalities (controls, n = 5). Laser-microdissected glomeruli were used for expression profiling of 762 miRNAs by low-density TaqMan arrays (cards A and B). The comparison of different normalization methods (GeNormPlus, NormFinder, global mean and snoRNAs) in crescentic IgA-GN, IgA-GN and controls yielded similar results. However, levels of significance and the range of relative expression differed. In median, two normalization methods demonstrated similar results. GeNormPlus and NormFinder gave different top ranked reference miRNAs. Stability ranking for snoRNAs varied between cards A and B. In conclusion, we suggest the geometric mean of the most stable reference miRNAs found in GeNormPlus (miR-26b-5p), NormFinder (miR-28-5p) and snoRNAs (RNU44) as reference. It should be considered that significant differences could be missed using one particular normalization method. As a starting point for glomerular miRNA studies in IgA nephropathy we provide a library of miRNAs.
Authors with CRIS profile
Sebastian Röder
Department of Neurology
Kerstin Ute Amann
Department of Nephropathology
Christoph Daniel
Department of Nephropathology
Involved external institutions
Universität Duisburg-Essen (UDE)
Germany (DE)
Universitätsklinikum Köln
Germany (DE)
Medizinische Hochschule Hannover (MHH) / Hannover Medical School
Germany (DE)
Germany (DE)
Universitätsklinikum Köln
Germany (DE)
Medizinische Hochschule Hannover (MHH) / Hannover Medical School
Germany (DE)
How to cite
APA:
Bockmeyer, C.L., Saeuberlich, K., Wittig, J., Esser, M., Röder, S., Vester, U.,... Becker, J.U. (2016). Comparison of different normalization strategies for the analysis of glomerular microRNAs in IgA nephropathy. Scientific Reports, 6, 31992. https://doi.org/10.1038/srep31992
MLA:
Bockmeyer, Clemens L., et al. "Comparison of different normalization strategies for the analysis of glomerular microRNAs in IgA nephropathy." Scientific Reports 6 (2016): 31992.
BibTeX: Download