title: Deep characterization of blood cell miRNomes by NGS.
abstract: A systematic understanding of different factors influencing cell type specific microRNA profiles is essential for state-of-the art biomarker research. We carried out a comprehensive analysis of the biological variability and changes in cell type pattern over time for different cell types and different isolation approaches in technical replicates. All combinations of the parameters mentioned above have been measured, resulting in 108 miRNA profiles that were evaluated by next-generation-sequencing. The largest miRNA variability was due to inter-individual differences (34 %), followed by the cell types (23.4 %) and the isolation technique (17.2 %). The change over time in cell miRNA composition was moderate (<3 %) being close to the technical variations (<1 %). Largest variability (including technical and biological variance) was observed for CD8 cells while CD3 and CD4 cells showed significantly lower variations. ANOVA highlighted that 51.5 % of all miRNAs were significantly influenced by the purification technique. While CD4 cells were least affected, especially miRNA profiles of CD8 cells were fluctuating depending on the cell purification approach. To provide researchers access to the profiles and to allow further analyses of the tested conditions we implemented a dynamic web resource.
authors.affiliation: Biophysics, Center for Integrative Physiology and Molecular Medicine, School of Medicine, Saarland University, Homburg, Germany.
authors.forename: Eva C
authors.initials: EC
authors.lastname: Schwarz
authors.affiliation: Saarland University, Building E2.1, 66123, Saarbrücken, Germany. c.backes@mx.uni-saarland.de.
authors.forename: Christina
authors.initials: C
authors.lastname: Backes
authors.affiliation: Biophysics, Center for Integrative Physiology and Molecular Medicine, School of Medicine, Saarland University, Homburg, Germany.
authors.forename: Arne
authors.initials: A
authors.lastname: Knörck
authors.affiliation: Department of Human Genetics, Saarland University, Homburg, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Ludwig
authors.affiliation: Department of Human Genetics, Saarland University, Homburg, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Leidinger
authors.affiliation: Biophysics, Center for Integrative Physiology and Molecular Medicine, School of Medicine, Saarland University, Homburg, Germany.
authors.forename: Cora
authors.initials: C
authors.lastname: Hoxha
authors.affiliation: Biophysics, Center for Integrative Physiology and Molecular Medicine, School of Medicine, Saarland University, Homburg, Germany.
authors.forename: Gertrud
authors.initials: G
authors.lastname: Schwär
authors.affiliation: Saarland University, Building E2.1, 66123, Saarbrücken, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Grossmann
authors.affiliation: Saarland University, Building E2.1, 66123, Saarbrücken, Germany.
authors.forename: Sabine C
authors.initials: SC
authors.lastname: Müller
authors.affiliation: Department of Human Genetics, Saarland University, Homburg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Hart
authors.affiliation: Department of Internal Medicine III, University Hospital Heidelberg, Heidelberg, Germany.|German Center for Cardiovascular Research (DZHK), Heidelberg, Germany.|Klaus Tschira Institute for Integrative Computational Cardiology, Heidelberg, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Haas
authors.affiliation: Saarland University, Building E2.1, 66123, Saarbrücken, Germany.
authors.forename: Valentina
authors.initials: V
authors.lastname: Galata
authors.affiliation: Clinical Hemostaseology and Transfusion Medicine, Saarland University, Homburg, Germany.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Müller
authors.affiliation: Saarland University, Building E2.1, 66123, Saarbrücken, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Fehlmann
authors.affiliation: Clinical Hemostaseology and Transfusion Medicine, Saarland University, Homburg, Germany.
authors.forename: Hermann
authors.initials: H
authors.lastname: Eichler
authors.affiliation: Kiel University, Kiel, Germany.
authors.forename: Andre
authors.initials: A
authors.lastname: Franke
authors.affiliation: Department of Internal Medicine III, University Hospital Heidelberg, Heidelberg, Germany.|German Center for Cardiovascular Research (DZHK), Heidelberg, Germany.|Klaus Tschira Institute for Integrative Computational Cardiology, Heidelberg, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Meder
authors.affiliation: Department of Human Genetics, Saarland University, Homburg, Germany.
authors.forename: Eckart
authors.initials: E
authors.lastname: Meese
authors.affiliation: Biophysics, Center for Integrative Physiology and Molecular Medicine, School of Medicine, Saarland University, Homburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Hoth
authors.affiliation: Saarland University, Building E2.1, 66123, Saarbrücken, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Keller
mesh_terms: D001483:Base Sequence; D001773:Blood Cells / Q000378:metabolism*; D016000:Cluster Analysis; D020869:Gene Expression Profiling / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000302:isolation & purification; D025341:Principal Component Analysis
chemical_list: D035683:MicroRNAs
keywords: Blood cells; FACS; Next-generation sequencing; microRNA