Abstract 4687: COL10A1, MMP-11 and ABHD2 expression in colorectal carcinoma primary tumors indicates metastatic disease.
Authors: Stürzl Michael , Wirtz Ralph , Konrad Andreas , Gefeller Olaf , Adler Werner , Naschberger Elisabeth , Prokosch Hans-Ullrich , Hartmann Arndt , Rau Tilman , Rödel Franz , Hohenberger Werner , Croner Roland S. 
Published: August 2014

Purpose: Metastasis is the primary cause of death in patients suffering from colorectal carcinomas (CRC). In order to identify patients at risk for metastatic tumor spread prior to clinical manifestation specific molecular markers are required. These markers will allow to adapt therapy early during clinical routine. Respective markers should be expressed in the primary tumor. In order to foster clinical implementation these markers should exhibit high validity and robust performance in routinely asservated material.

Patients and Methods: Five metastasis associated markers (ABDH2, COL10A1, MMP-11, C8orf30A, SLC35D1) were identified by microarray comparison of non-metastatic and metastatic fresh frozen CRC samples (n = 80). They were validated by TaqMan quantitative RT-PCR in two further independent patient cohorts either retrospectively (n = 82) or prospectively (n = 203). RNA for validation was isolated with a fully automated procedure (Tissue Preparation System, Siemens Healthcare Diagnostics) from routinely harvested formaldehyde-fixed, paraffin-embedded (FFPE) CRC tissues. Relation of marker expression to disease recurrence was analyzed in UICC II stage tumors. Prognostic power was evaluated by different multiparametric tests (Bagging, SVM, LDA). Results were controlled with immunohistochemistry at the protein level.

Results: ABDH2, COL10A1, MMP-11, C8orf30A and SLC35D1 were significantly differently expressed in stage UICC I, II vs. III by microarray analyses (p&lt;0.001). Retrospective and prospective RT-PCR validation in FFPE tissues confirmed that COL10A1, MMP-11, and ABHD2 were robustly expressed and significantly (p &lt; 0.001) associated with metastasis (stage UICC III/VI). Analyses of follow up in stage UICC II CRC patients indicated a relation to tumor recurrence. Multiparametric tests identified AUC values between 0.714 (Bagged trees) and 0.736 (LDA) for these markers. Controlling of the results with immunohistochemistry demonstrated that high prognostic power was selectively obtained at the RNA expression level.

Conclusion: Automated RNA extraction from FFPE tissues and subsequent qRT-PCR analysis provide a robust technology which can be implemented into clinical routine processes of molecular cancer diagnostics. COL10A1, MMP-11 and ABHD2 are robustly expressed in a metastasis-related manner in CRC primary tumors. All three markers may be used as prognostic profilers to identify high risk patients.

Citation Format: Michael Stürzl, Ralph Wirtz, Andreas Konrad, Olaf Gefeller, Werner Adler, Elisabeth Naschberger, Hans-Ullrich Prokosch, Arndt Hartmann, Tilman Rau, Franz Rödel, Werner Hohenberger, Roland S. Croner. COL10A1, MMP-11 and ABHD2 expression in colorectal carcinoma primary tumors indicates metastatic disease. [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 4687. doi:10.1158/1538-7445.AM2013-4687