Differentially expressed mRNA/proteins can distinguish viable germ cell tumors and teratomas from necrosis in retroperitoneal lymph node resections after chemotherapy (pcRPLND).

Authors

null

Tim Nestler

Department of Urology and Uro-Oncology, University Hospital of Cologne, Cologne, Germany

Tim Nestler , Lara Kremer , Svenja Wagener-Ryczek , Maike Wittersheim , Melanie von Brandenstein , Pia Paffenholz , Stefan Mueller , Alexander Quaas , Martin Hellmich , Margarete Odenthal , David Pfister , Axel Heidenreich

Organizations

Department of Urology and Uro-Oncology, University Hospital of Cologne, Cologne, Germany, Institute of Pathology, University of Cologne, Cologne, Germany, Institute of Pathology, University Hospital of Cologne, Cologne, Germany, Center for Molecular Medicine Cologne (CMMC), University of Cologne, Cologne, Germany, Institute of Medical Statistics and Computational Biology, University of Cologne, Cologne, Germany, University Hospital of Cologne, Cologne, Germany

Research Funding

Other

Background: Metastatic non-seminomatous testicular tumor patients with residual retroperitoneal tumor masses > 1cm after chemotherapy are treated with pcRPLND. The goal of pcRPLND is to remove viable tumors (V) and teratoma (T), which are present in approximately 10% and 40% of cases, respectively. However, histopathologically, only scar/necrosis (N) is identified in the remaining 50% of patients. In those patients, surgical therapy is not necessary, resulting in a relevant overtreatment. So far, no adequate distinction between the histologies exists preoperatively. Recently, the first biomarker was described with miR371a-3p in serum, which is highly specific for V, but not for T. Therefore, our goal was to identify mRNAs and proteins that are differentially expressed between V/T vs N, in particular between T and N, in pcRPLND resected cells. Methods: Forty-eight patients were identified, n = 16 each with T/V/N. Representative regions of T/V/N were microdissected and subsequently mRNA was extracted. Initially, 770 genes were analyzed using the nCounter PanCancer Progression Panel (Nanostring). For each group comparison, genes with a fold change of < -2/ > 2 and a p-value of < 0.05 were identified. Hereafter, quantitative protein analysis (proteomics) was performed on the same samples. Finally, the proteins of the 5 mRNAs with the most different and significant expression levels between T vs. N were validated by immunohistochemistry and H-score calculation. Results: By Nanostring, we identified 84 significantly differentially expressed mRNAs for the group comparisons of T vs. N, 63 for V vs. N, and 189 for T vs. V. Quantitative protein analysis revealed 25 significantly differentially expressed proteins in T vs. N, 254 in V vs. N, and 134 between T vs. V. By immunohistochemistry, all 5 antibodies showed significantly increased H scores when comparing T vs. N and T vs. V. In accordance with the objective, we found two proteins, AGR2 and KRT19, with their corresponding genes that showed significantly differential expressions for the comparison of T vs. N in both, quantitative protein analysis and Nanostring mRNA analysis, and were successfully validated by immunohistochemistry. Conclusions: With AGR2 and KRT19, we have identified two proteins with their corresponding genes that are significantly and differentially expressed in the pcRPLND specimen in the clinically relevant groups T vs. N. Both were successfully validated by immunohistochemistry. In addition, further group differences (T vs. V/ V vs. N) were revealed depending on the analytical method. In perspective, these proteins could be targeted by radiolabeled ligands as a tracer in order to reliably distinguish patients with teratoma from those with necrosis by means of functional imaging. Thus, overtreatment with pcRPLND of patients with N could be safely reduced.

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Abstract Details

Meeting

2022 ASCO Genitourinary Cancers Symposium

Session Type

Poster Session

Session Title

Poster Session C: Renal Cell Cancer; Adrenal, Penile, Urethral, and Testicular Cancers

Track

Renal Cell Cancer,Adrenal Cancer,Penile Cancer,Testicular Cancer,Urethral Cancer

Sub Track

Translational Research, Tumor Biology, Biomarkers, and Pathology

Citation

J Clin Oncol 40, 2022 (suppl 6; abstr 408)

DOI

10.1200/JCO.2022.40.6_suppl.408

Abstract #

408

Poster Bd #

D1

Abstract Disclosures

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