title: Targeted serum miRNA (TSmiR) test for diagnosis and follow-up of (testicular) germ cell cancer patients: a proof of principle.
abstract: Germ cell cancers (GCC) are the most frequent malignancy in young Caucasian males. GCC can consist of seminomas (SE) and non-seminomas (malignant NS: embryonal carcinoma (EC), yolk sac tumor (YS), choriocarcinoma (CH) and teratoma (TE)). Current serum-markers used for diagnosis and follow-up (AFP, hCG) are predominantly related to YS and CH and marker positivity can vary during disease. Therefore, stable markers consistently identifying more GCC components, specifically the stem cell components SE and EC, are of interest. Expression of the embryonic stem cell miR-371-3 and miR-302/367 clusters in SE/EC/YS suggest possible application of these micro-RNAs as GCC tumor-markers. The TSmiR protocol constitutes a complete, quality-controlled pipeline for the detection of miRs in serum, based on magnetic bead-based purification and qPCR quantification. As a proof of principle, TSmiR was applied to five independent serum sample series including 80 GCCs, 47 controls, 11 matched pre/post orchidectomy samples and 12 no-GCC testicular masses. GCC serum samples showed a consistent, significant (p < 0.0064) increase of miR-371/372/373/367 levels. Analogous, serum levels returned to baseline after orchidectomy (stage-I disease). Moreover, there was a trend toward higher miR levels in patients with metastasis. These results imply suitability for diagnosis and follow-up. TSmiR showed an overall sensitivity of 98%, clearly outperforming the traditional serum markers AFP/hCG (36%/57%, sensitivity(AFP) = 3%/45%; sensitivity(hCG) = 62%/66%, SE/NS). TSmiR misclassified one tumor as a control. Serum AFP/hCG and TSmiR combined identified all T samples correctly. In conclusion, TSmiR constitutes a highly sensitive and reproducible serum test for GCC patients, suitable to be prospectively tested for diagnostic and follow-up purposes.
authors.affiliation: Department of Pathology, Erasmus MC - University Medical Center Rotterdam, Building Be, Room 432, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Ad J M
authors.initials: AJ
authors.lastname: Gillis
authors.forename: Martin A
authors.initials: MA
authors.lastname: Rijlaarsdam
authors.forename: Ronak
authors.initials: R
authors.lastname: Eini
authors.forename: Lambert C J
authors.initials: LC
authors.lastname: Dorssers
authors.forename: Katharina
authors.initials: K
authors.lastname: Biermann
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Murray
authors.forename: James C
authors.initials: JC
authors.lastname: Nicholson
authors.forename: Nicholas
authors.initials: N
authors.lastname: Coleman
authors.forename: Klaus-Peter
authors.initials: KP
authors.lastname: Dieckmann
authors.forename: Gazanfer
authors.initials: G
authors.lastname: Belge
authors.forename: Jörn
authors.initials: J
authors.lastname: Bullerdiek
authors.forename: Tom
authors.initials: T
authors.lastname: Xu
authors.forename: Nathalie
authors.initials: N
authors.lastname: Bernard
authors.forename: Leendert H J
authors.initials: LH
authors.lastname: Looijenga
mesh_terms: D000293:Adolescent; D000328:Adult; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D002648:Child; D002675:Child, Preschool; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009373:Neoplasms, Germ Cell and Embryonal / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism*; D013736:Testicular Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism* / Q000473:pathology
chemical_list: D014408:Biomarkers, Tumor; D035683:MicroRNAs; D012334:RNA, Neoplasm
keywords: AFP; Germ cell cancer; Serum markers; Testicular cancer; hCG; miRNA