Abstract 5321: An informative clinical applicable serum miRNAs 371-3 and 302/367 test for (germ cell) cancer patients.
Authors: Gillis Ad , Rijlaarsdam Martin , Eini Ronak , Dorssers Lambert , van der Zwan Yvonne , Murray Matthew , Nicholson James , Coleman Nicholas , Dieckmann Klaus-Peter , Belge Ganzafar , Bullerdieck Joern , Biermann Katharina , White Stefan , Looijenga Leendert H. 
Published: August 2014

Expression of microRNAs (miR) can be unique and specific for malignant cells, as shown for human germ cell cancers (GCC), i.e., seminomas (SE) and nonseminomas (NS). GCC are the most frequent malignancy in Caucasian young males. They show high expression of the embryonic stem cell miR clusters 371-3 and 302/367, related to their cancer stem cell component. No recurrent DNA copy number changes of these miR and mutation in DICER was found in more than 250 primary GCC samples. Current clinically used serum markers for GCC are the proteins AFP and hCG, which are informative in a selection of patients (minority with a SE, and majority of those with a NS: i.p. yolk sac tumor and/or choriocarcinoma). Although the fore-mentioned miR have been suggested as serum markers for GCC, no robust protocol is available so far. A novel, sensitive and specific pipeline for miRNA profiling in body fluids is developed based on a magnetic anti-probe bead purification step (ABC miRNA purification kit, Life Technologies). This method shows stable recovery based on multiple synthetic non-human spike-ins, allowing calibration. Out of a set of 9 potential serum-specific controls, two were identified by Normfinder and geNorm as most stable (miR-20a and 93), therefore used for normalization. This TSmiR test was applied to five series of serum samples for independent learning and validation, including 47 controls (N), 80 GCC (adult and pediatric) (T), 11 matched pre- and post-orchidectomy samples, and 12 no-GCC testicular tumors (no-GCC). TSmiR demonstrated a consistent, significant increase (p&lt;0.0064) of miR-371-3 and 367 levels in serum of patients with GCC. Power was demonstrated using mean comparison, Kohonen Self Organizing Maps, Principal Component Analysis and ROC analysis. Matched pre- and post-orchidectomy samples demonstrated suitability for follow-up purposes. TSmiR outperformed the traditional serum markers (sensitivityall samples 98%, specificityall samples 48%; sensitivityAFP= 3% and 45%; sensitivityhCG= 62% and 66%, in SE and NS, respectively). The combined serum AFP/hCG and TSmiR test identified all T samples correctly. In conclusion, a highly reproducible and informative novel serum test for patients with a germ cell malignancy is developed, suitable to be prospectively tested for diagnosis and follow-up purposes.

Citation Format: Ad Gillis, Martin Rijlaarsdam, Ronak Eini, Lambert Dorssers, Yvonne van der Zwan, Matthew Murray, James Nicholson, Nicholas Coleman, Klaus-Peter Dieckmann, Ganzafar Belge, Joern Bullerdieck, Katharina Biermann, Stefan White, Leendert H. Looijenga. An informative clinical applicable serum miRNAs 371-3 and 302/367 test for (germ cell) cancer patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5321. doi:10.1158/1538-7445.AM2013-5321