title: Towards a new standardized method for circulating miRNAs profiling in clinical studies: Interest of the exogenous normalization to improve miRNA signature accuracy.
abstract: Circulating miRNAs are promising biomarkers in oncology but have not yet been implemented in the clinic given the lack of concordance across studies. In order to increase the cross-studies reliability, we attempted to reduce and to control the circulating miRNA expression variability between patients. First, to maximize profiling signals and to reduce miRNA expression variability, three isolation kits were compared and the NucleoSpin(®) kit provided higher miRNA concentrations than the other widely used kits. Second, to control inter-sample variability during the profiling step, the exogenous miRNAs normalization method commonly used for RT-qPCR validation step was adapted to microarray experiments. Importantly, exogenous miRNAs presented two-fold lower inter-sample variability than the widely used endogenous miR-16-5p reflecting that the latter is subject to both biological and technical variability. Although Caenorhabditis elegans miRNAs isolation yields were heterogeneous, they correlated to each other and to their geometrical mean across samples. The normalization based on the geometrical mean of three exogenous miRNAs increased the correlation up-to 0.97 between the microarrays and individual RT-qPCR steps of circulating miRNAs expression. Overall, this new strategy open new avenue to identify reliable circulating miRNA signatures for translation into clinical practice.
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Vigneron
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Meryet-Figuière
authors.affiliation: EPHE, Inserm U823, CaCys, Grenoble, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Guttin
authors.affiliation: Inserm U1216, Transcriptomics Core Facility, Grenoble, France; Univ. Grenoble Alpes, GIN, Grenoble, France.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Issartel
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France; CNRS, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Lambert
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France.
authors.forename: Mélanie
authors.initials: M
authors.lastname: Briand
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France.
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: Louis
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France.
authors.forename: Mégane
authors.initials: M
authors.lastname: Vernon
authors.affiliation: Inserm U1086, Cancers & Preventions, Caen, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Lebailly
authors.affiliation: Inserm U1086, Cancers & Preventions, Caen, France.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Lecluse
authors.affiliation: Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France; Inserm U1086, Cancers & Preventions, Caen, France; Medical Oncology Department, Comprehensive Cancer Center F. Baclesse, Caen, France; Clinical Research Department, Comprehensive Cancer Center F. Baclesse, Caen, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Joly
authors.affiliation: Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France; Biopathology Department, Comprehensive Cancer Center F. Baclesse, Caen, France; Inserm U1079, Institute for Research and Innovation in Biomedicine, University of Rouen, Rouen, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Krieger
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France; Medical Oncology Department, Comprehensive Cancer Center F. Baclesse, Caen, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Lheureux
authors.affiliation: Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France; Clinical Research Department, Comprehensive Cancer Center F. Baclesse, Caen, France.
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Clarisse
authors.affiliation: Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France; Clinical Research Department, Comprehensive Cancer Center F. Baclesse, Caen, France.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Leconte
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France.
authors.forename: Pascal
authors.initials: P
authors.lastname: Gauduchon
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Poulain
authors.affiliation: Inserm U1199, « Biology and Innovative Therapeutics for Locally Aggressive Cancer » (BioTICLA) Unit, Caen, France; Normandie Univ, France; UNICAEN, France; Comprehensive Cancer Center F. Baclesse, Unicancer, Caen, France. Electronic address: c.denoyelle@baclesse.unicancer.fr.
authors.forename: Christophe
authors.initials: C
authors.lastname: Denoyelle
mesh_terms: D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D035683:MicroRNAs / Q000097:blood* / Q000235:genetics*; D009369:Neoplasms / Q000097:blood / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards
chemical_list: D035683:MicroRNAs
keywords: Circulating miRNA; Normalization; Ovarian cancer; Plasma; Profiling; Serum