miRNA expression profiling enables risk stratification in archived and fresh neuroblastoma tumor samples.
OBJECTIVE
More accurate assessment of prognosis is important to further improve the choice of risk-related therapy in neuroblastoma (NB) patients. In this study, we aimed to establish and validate a prognostic miRNA signature for children with NB and tested it in both fresh frozen and archived formalin-fixed paraffin-embedded (FFPE) samples.


METHODS
Four hundred-thirty human mature miRNAs were profiled in two patient subgroups with maximally divergent clinical courses. Univariate logistic regression analysis was used to select miRNAs correlating with NB patient survival. A 25-miRNA gene signature was built using 51 training samples, tested on 179 test samples, and validated on an independent set of 304 fresh frozen tumor samples and 75 archived FFPE samples.


RESULTS
The 25-miRNA signature significantly discriminates the test patients with respect to progression-free and overall survival (P < 0.0001), both in the overall population and in the cohort of high-risk patients. Multivariate analysis indicates that the miRNA signature is an independent predictor of patient survival after controlling for current risk factors. The results were confirmed in an external validation set. In contrast to a previously published mRNA classifier, the 25-miRNA signature was found to be predictive for patient survival in a set of 75 FFPE neuroblastoma samples.


CONCLUSIONS
In this study, we present the largest NB miRNA expression study so far, including more than 500 NB patients. We established and validated a robust miRNA classifier, able to identify a cohort of high-risk NB patients at greater risk for adverse outcome using both fresh frozen and archived material.
Center for Medical Genetics; Department of Paediatric Hematology and Oncology, Ghent University Hospital, Ghent, Belgium.
Katleen
K
De Preter
Pieter
P
Mestdagh
Joëlle
J
Vermeulen
Fjoralba
F
Zeka
Arlene
A
Naranjo
Isabella
I
Bray
Victoria
V
Castel
Caifu
C
Chen
Elzbieta
E
Drozynska
Angelika
A
Eggert
Michael D
MD
Hogarty
Ewa
E
Izycka-Swieszewska
Wendy B
WB
London
Rosa
R
Noguera
Marta
M
Piqueras
Kenneth
K
Bryan
Benjamin
B
Schowe
Peter
P
van Sluis
Jan J
JJ
Molenaar
Alexander
A
Schramm
Johannes H
JH
Schulte
Raymond L
RL
Stallings
Rogier
R
Versteeg
Geneviève
G
Laureys
Nadine
N
Van Roy
Frank
F
Speleman
Jo
J
Vandesompele
D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D015331:Cohort Studies; D005500:Follow-Up Studies; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D007223:Infant; D053208:Kaplan-Meier Estimate; D016015:Logistic Models; D035683:MicroRNAs / Q000235:genetics*; D015999:Multivariate Analysis; D009447:Neuroblastoma / Q000175:diagnosis* / Q000235:genetics* / Q000628:therapy; D011379:Prognosis; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018570:Risk Assessment; D012307:Risk Factors; D013997:Time Factors
D035683:MicroRNAs