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Br J Cancer. 2016 Jan 19;114(2):151-62. doi: 10.1038/bjc.2015.429. Epub 2015 Dec 15.

A pipeline to quantify serum and cerebrospinal fluid microRNAs for diagnosis and detection of relapse in paediatric malignant germ-cell tumours.

Author information

1
Department of Paediatric Haematology and Oncology, Addenbrooke's Hospital, Hills Road, Cambridge CB2 0QQ, UK.
2
Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK.
3
Department of Paediatrics, University of Cambridge, Addenbrooke's Hospital, Cambridge CB2 0QQ, UK.
4
AstraZeneca, Cambridge Science Park, Cambridge CB4 0FZ, UK.
5
Department of Pathology, Erasmus MC University Medical Center Rotterdam, Cancer Institute, Rotterdam, Netherlands.
6
Department of Histopathology, Addenbrooke's Hospital, Hills Road, Cambridge CB2 0QQ, UK.

Abstract

BACKGROUND:

The current biomarkers alpha-fetoprotein and human chorionic gonadotropin have limited sensitivity and specificity for diagnosing malignant germ-cell tumours (GCTs). MicroRNAs (miRNAs) from the miR-371-373 and miR-302/367 clusters are overexpressed in all malignant GCTs, and some of these miRNAs show elevated serum levels at diagnosis. Here, we developed a robust technical pipeline to quantify these miRNAs in the serum and cerebrospinal fluid (CSF). The pipeline was used in samples from a cohort of exclusively paediatric patients with gonadal and extragonadal malignant GCTs, compared with appropriate tumour and non-tumour control groups.

METHODS:

We developed a method for miRNA quantification that enabled sample adequacy assessment and reliable data normalisation. We performed qRT-PCR profiling for miR-371-373 and miR-302/367 cluster miRNAs in a total of 45 serum and CSF samples, obtained from 25 paediatric patients.

RESULTS:

The exogenous non-human spike-in cel-miR-39-3p and the endogenous housekeeper miR-30b-5p were optimal for obtaining robust serum and CSF qRT-PCR quantification. A four-serum miRNA panel (miR-371a-3p, miR-372-3p, miR-373-3p and miR-367-3p): (i) showed high sensitivity/specificity for diagnosing paediatric extracranial malignant GCT; (ii) allowed early detection of relapse of a testicular mixed malignant GCT; and (iii) distinguished intracranial malignant GCT from intracranial non-GCT tumours at diagnosis, using CSF and serum samples.

CONCLUSIONS:

The pipeline we have developed is robust, scalable and transferable. It potentially promises to improve clinical management of paediatric (and adult) malignant GCTs.

PMID:
26671749
PMCID:
PMC4815809
DOI:
10.1038/bjc.2015.429
[Indexed for MEDLINE]
Free PMC Article

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