In this study, the circulating miRNome from diagnostic neuroblastoma serum was assessed for identification of noninvasive biomarkers with potential in monitoring metastatic disease. After determining the circulating neuroblastoma miRNome, 743 miRNAs were screened in 2 independent cohorts of 131 and 54 patients. Evaluation of serum miRNA variance in a model testing for tumor stage, MYCN status, age at diagnosis, and overall survival revealed tumor stage as the most significant factor impacting miRNA abundance in neuroblastoma serum. Differential abundance analysis between patients with metastatic and localized disease revealed 9 miRNAs strongly associated with metastatic stage 4 disease in both patient cohorts. Increasing levels of these miRNAs were also observed in serum from xenografted mice bearing human neuroblastoma tumors. Moreover, murine serum miRNA levels were strongly associated with tumor volume. These findings were validated in longitudinal serum samples from metastatic neuroblastoma patients, where the 9 miRNAs were associated with disease burden and treatment response.

Cancer Research Institute Ghent Ghent University Ghent Belgium

Center for Medical Genetics Department of Biomolecular Medicine and

Center for Molecular Medicine Cologne University of Cologne Cologne Germany

Department of Paediatric Oncology Institut Curie Paris France

Department of Pathology Medical School University of Valencia CIBERONC Madrid Spain

Department of Pediatric Hematology Oncology and Stem Cell Transplantation Ghent University Hospital Ghent Belgium

Department of Pediatric Oncology University Hospital Brno Brno Czech Republic

Department of Pediatrics Section of Hematology Oncology Texas Children's Cancer Center Baylor College of Medicine Houston Texas USA

Division of Oncology The Children's Hospital of Philadelphia Philadelphia USA

Genome Informatics Institute of Human Genetics University Hospital Essen University of Duisburg Essen Essen Germany

German Cancer Research Center Heidelberg Germany

INCLIVA Biomedical Research Institute Valencia Spain

Laboratoire de Recherche Translationnelle Centre Léon Bérard Lyon France

Molecular Oncology West German Cancer Center University Hospital Essen University of Duisburg Essen Essen Germany

Pediatric Hematology Oncology Schneider Children's Medical Center of Israel Tel Aviv Israel

Pediatric Oncology and Hematology Charité University Medicine Berlin Germany

Pediatric Oncology and Hematology University Children's Hospital of Cologne Medical Faculty and

Pédiatrie Hôpital de Jolimont Haine Saint Paul Belgium

Zobrazit více v PubMed

Cheung NK, Dyer MA. Neuroblastoma: developmental biology, cancer genomics and immunotherapy. Nat Rev Cancer. 2013;13(6):397–411. doi: 10.1038/nrc3526. PubMed DOI PMC

Smith MA, Altekruse SF, Adamson PC, Reaman GH, Seibel NL. Declining childhood and adolescent cancer mortality. Cancer. 2014;120(16):2497–2506. doi: 10.1002/cncr.28748. PubMed DOI PMC

Hartomo TB, et al. Minimal residual disease monitoring in neuroblastoma patients based on the expression of a set of real-time RT-PCR markers in tumor-initiating cells. Oncol Rep. 2013;29(4):1629–1636. doi: 10.3892/or.2013.2286. PubMed DOI

Pugh TJ, et al. The genetic landscape of high-risk neuroblastoma. Nat Genet. 2013;45(3):279–284. doi: 10.1038/ng.2529. PubMed DOI PMC

Blondal T, et al. Assessing sample and miRNA profile quality in serum and plasma or other biofluids. Methods. 2013;59(1):S1–S6. doi: 10.1016/j.ymeth.2012.09.015. PubMed DOI

Murray MJ, et al. Solid tumors of childhood display specific serum microRNA profiles. Cancer Epidemiol Biomarkers Prev. 2015;24(2):350–360. doi: 10.1158/1055-9965.EPI-14-0669. PubMed DOI PMC

Ramraj SK, Aravindan S, Somasundaram DB, Herman TS, Natarajan M, Aravindan N. Serum-circulating miRNAs predict neuroblastoma progression in mouse model of high-risk metastatic disease. Oncotarget. 2016;7(14):18605–18619. PubMed PMC

Mestdagh P, et al. High-throughput stem-loop RT-qPCR miRNA expression profiling using minute amounts of input RNA. Nucleic Acids Res. 2008;36(21):e143. doi: 10.1093/nar/gkn725. PubMed DOI PMC

Schulte JH, et al. Deep sequencing reveals differential expression of microRNAs in favorable versus unfavorable neuroblastoma. Nucleic Acids Res. 2010;38(17):5919–5928. doi: 10.1093/nar/gkq342. PubMed DOI PMC

Chicard M, et al. Genomic Copy Number Profiling Using Circulating Free Tumor DNA Highlights Heterogeneity in Neuroblastoma. Clin Cancer Res. 2016;22(22):5564–5573. doi: 10.1158/1078-0432.CCR-16-0500. PubMed DOI

Van Roy N, et al. Shallow Whole Genome Sequencing on Circulating Cell-Free DNA Allows Reliable Noninvasive Copy-Number Profiling in Neuroblastoma Patients. Clin Cancer Res. 2017;23(20):6305–6314. doi: 10.1158/1078-0432.CCR-17-0675. PubMed DOI

Combaret V, et al. Circulating MYCN DNA as a tumor-specific marker in neuroblastoma patients. Cancer Res. 2002;62(13):3646–3648. PubMed

Zeka F, Mestdagh P, Vandesompele J. RT-qPCR-based quantification of small non-coding RNAs. Methods Mol Biol. 2015;1296:85–102. doi: 10.1007/978-1-4939-2547-6_9. PubMed DOI

Mestdagh P, et al. Evaluation of quantitative miRNA expression platforms in the microRNA quality control (miRQC) study. Nat Methods. 2014;11(8):809–815. doi: 10.1038/nmeth.3014. PubMed DOI

Agarwal S, et al. Transmembrane adaptor protein PAG1 is a novel tumor suppressor in neuroblastoma. Oncotarget. 2016;7(17):24018–24026. PubMed PMC

Van Goethem A, et al. Depletion of tRNA-halves enables effective small RNA sequencing of low-input murine serum samples. Sci Rep. 2016;6:37876. PubMed PMC

D’haene B, Mestdagh P, Hellemans J, Vandesompele J. miRNA expression profiling: from reference genes to global mean normalization. Methods Mol Biol. 2012;822:261–272. doi: 10.1007/978-1-61779-427-8_18. PubMed DOI

Wood. Generalized Additive Models: An Introduction with R. Second Edition. Boca Ratan, FL:Chapman and Hall/CRC;2017.

Hellemans J, Mortier G, De Paepe A, Speleman F, Vandesompele J. qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol. 2007;8(2):R19. doi: 10.1186/gb-2007-8-2-r19. PubMed DOI PMC