Comparison of different normalization strategies for the analysis of glomerular microRNAs in IgA nephropathy View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-10

AUTHORS

Clemens L Bockmeyer, Karen Säuberlich, Juliane Wittig, Marc Eßer, Sebastian S Roeder, Udo Vester, Peter F Hoyer, Putri A Agustian, Philip Zeuschner, Kerstin Amann, Christoph Daniel, Jan U Becker

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. More... »

PAGES

31992

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/srep31992

DOI

http://dx.doi.org/10.1038/srep31992

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1022256759

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/27553688


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