A pipeline to quantify serum and cerebrospinal fluid microRNAs for diagnosis and detection of relapse in paediatric malignant germ cell tumours
Raby, Katie L
Rijlaarsdam, Martin A
Gillis, Ad JM
Looijenga, Leendert HJ
Nicholson, James C
Serum and CSF microRNAs in paediatric malignant GCTs
British Journal of Cancer
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Murray, M., Bell, E., Raby, K. L., Rijlaarsdam, M. A., Gillis, A. J., Looijenga, L. H., Brown, H., et al. (2015). A pipeline to quantify serum and cerebrospinal fluid microRNAs for diagnosis and detection of relapse in paediatric malignant germ cell tumours. British Journal of Cancer, 114 151-162. https://doi.org/10.1038/bjc.2015.429
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 over-expressed 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 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; 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.
biomarker, blood, diagnosis, germ cell tumour, microRNA, paediatric, relapse, serum
Grant funding was from CwCUK/GOSHCC (M.J. Murray, K.L. Raby, J.C. Nicholson, N. Coleman), SPARKS (M.J. Murray, J.C. Nicholson, N. Coleman), AstraZeneca (E. Bell, H. Brown and B. Destenaves), CRUK (N. Coleman), MRC (M.J. Murray) and an Erasmus MC MRACE grant (M.A. Rijlaarsdam). The authors also thank the Max Williamson Fund and The Perse Preparatory School, Cambridge for supporting this study.
External DOI: https://doi.org/10.1038/bjc.2015.429
This record's URL: https://www.repository.cam.ac.uk/handle/1810/252635