Universität zu Köln

Bockmeyer, Clemens L., Saeuberlich, Karen, Wittig, Juliane, Esser, Marc, Roeder, Sebastian S., Vester, Udo, Hoyer, Peter F., Agustian, Putri A., Zeuschner, Philip ORCID: 0000-0001-8898-6588, Amann, Kerstin, Daniel, Christoph and Becker, Jan U. (2016). Comparison of different normalization strategies for the analysis of glomerular microRNAs in IgA nephropathy. Sci Rep, 6. LONDON: NATURE PUBLISHING GROUP. ISSN 2045-2322

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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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Bockmeyer, Clemens L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Saeuberlich, Karen UNSPECIFIED UNSPECIFIED UNSPECIFIED Wittig, Juliane UNSPECIFIED UNSPECIFIED UNSPECIFIED Esser, Marc UNSPECIFIED UNSPECIFIED UNSPECIFIED Roeder, Sebastian S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Vester, Udo UNSPECIFIED UNSPECIFIED UNSPECIFIED Hoyer, Peter F. UNSPECIFIED UNSPECIFIED UNSPECIFIED Agustian, Putri A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Zeuschner, Philip UNSPECIFIED orcid.org/0000-0001-8898-6588 UNSPECIFIED Amann, Kerstin UNSPECIFIED UNSPECIFIED UNSPECIFIED Daniel, Christoph UNSPECIFIED UNSPECIFIED UNSPECIFIED Becker, Jan U. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-266317
DOI:	10.1038/srep31992
Journal or Publication Title:	Sci Rep
Volume:	6
Date:	2016
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	2045-2322
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DIABETIC-NEPHROPATHY; MESANGIAL CELLS; DOWN-REGULATION; REFERENCE GENES; MESSENGER-RNA; PCR DATA; RELATIVE QUANTIFICATION; QUANTITATIVE PCR; KIDNEY-DISEASE; EXPRESSION Multiple languages Multidisciplinary Sciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/26631