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Moving towards Personalized Medicine in Muscle-Invasive Bladder Cancer: Where Are We Now and Where Are We Going?
Open AccessArticle

Prognostic Role of Survivin and Macrophage Infiltration Quantified on Protein and mRNA Level in Molecular Subtypes Determined by RT-qPCR of KRT5, KRT20, and ERBB2 in Muscle-Invasive Bladder Cancer Treated by Adjuvant Chemotherapy

1
Department of Urology, HELIOS Hospital Bad Saarow, DE-15526 Bad Sarrow, Germany
2
Brandenburg Medical School, DE-14770 Brandenburg, Germany
3
Department of Urology, Charité—Universitätsmedizin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, DE-10098 Berlin, Germany
4
Berlin Institute for Urological Research, DE-10098 Berlin, Germany
5
Institute of Pathology, DE-51375 Leverkusen, Germany
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Department of Urology, Turku University Hospital, FI-20521 Turku, Finland
7
MediCity Research Laboratory, Department of Medical Microbiology and Immunology, University of Turku, FI-20520 Turku, Finland
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Institute of Pathology, Turku University Hospital, FI-20521 Turku, Finland
9
Institute for Prevention and Occupational Medicine of the German Social Accident Insurance (IPA), Institute of the Ruhr University Bochum, DE-44789 Bochum, Germany
10
Institute of Pathology, HELIOS Hospital Bad Saarow, DE-15526 Bad Sarrow, Germany
11
STRATIFYER Molecular Pathology GmbH, DE-50935 Cologne, Germany
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(19), 7420; https://doi.org/10.3390/ijms21197420
Received: 29 July 2020 / Revised: 18 September 2020 / Accepted: 30 September 2020 / Published: 8 October 2020
Objectives: Bladder cancer is a heterogeneous malignancy. Therefore, it is difficult to find single predictive markers. Moreover, most studies focus on either the immunohistochemical or molecular assessment of tumor tissues by next-generation sequencing (NGS) or PCR, while a combination of immunohistochemistry (IHC) and PCR for tumor marker assessment might have the strongest impact to predict outcome and select optimal therapies in real-world application. We investigated the role of proliferation survivin/BIRC5 and macrophage infiltration (CD68, MAC387, CLEVER-1) on the basis of molecular subtypes of bladder cancer (KRT5, KRT20, ERBB2) to predict outcomes of adjuvant treated muscle-invasive bladder cancer patients with regard to progression-free survival (PFS) and disease-specific survival (DSS). Materials and Methods: We used tissue microarrays (TMA) from n = 50 patients (38 males, 12 female) with muscle-invasive bladder cancer. All patients had been treated with radical cystectomy followed by adjuvant triple chemotherapy. Median follow-up time was 60.5 months. CD68, CLEVER-1, MAC387, and survivin protein were detected by immunostaining and subsequent visual inspection. BIRC5, KRT5, KRT20, ERBB2, and CD68 mRNAs were detected by standardized RT-qPCR after tissue dot RNA extraction using a novel stamp technology. All these markers were evaluated in three different centers of excellence. Results: Nuclear staining rather than cytoplasmic staining of survivin predicted DSS as a single marker with high levels of survivin being associated with better PFS and DSS upon adjuvant chemotherapy (p = 0.0138 and p = 0.001, respectively). These results were validated by the quantitation of BIRC5 mRNA by PCR (p = 0.0004 and p = 0.0508, respectively). Interestingly, nuclear staining of survivin protein was positively associated with BIRC5 mRNA, while cytoplasmic staining was inversely related, indicating that the translocation of survivin protein into the nucleus occurred at a discrete, higher level of its mRNA. Combining survivin/BIRC5 levels based on molecular subtype being assessed by KRT20 expression improved the predictive value, with tumors having low survivin/BIRC5 and KRT20 mRNA levels having the best survival (75% vs. 20% vs. 10% 5-year DSS, p = 0.0005), and these values were independent of grading, node status, and tumor stage in multivariate analysis (p = 0.0167). Macrophage infiltration dominated in basal tumors and was inversely related with the luminal subtype marker gene expression. The presence of macrophages in survivin-positive or ERBB2-positive tumors was associated with worse DSS. Conclusions: For muscle-invasive bladder cancer patients, the proliferative activity as determined by the nuclear staining of survivin or RT-qPCR on the basis of molecular subtype characteristics outperforms single marker detections and single technology approaches. Infiltration by macrophages detected by IHC or PCR is associated with worse outcome in defined subsets of tumors. The limitations of this study are the retrospective nature and the limited number of patients. However, the number of molecular markers has been restricted and based on predefined assumptions, which resulted in the dissection of muscle-invasive disease into tumor–biological axes of high prognostic relevance, which warrant further investigation and validation. View Full-Text
Keywords: survivin; BIRC5; macrophage; KRT20; ERBB2; MIBC; prediction; RT-qPCR; adjuvant chemotherapy; survival; bladder cancer survivin; BIRC5; macrophage; KRT20; ERBB2; MIBC; prediction; RT-qPCR; adjuvant chemotherapy; survival; bladder cancer
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Ecke, T.H.; Kiani, A.; Schlomm, T.; Friedersdorff, F.; Rabien, A.; Jung, K.; Kilic, E.; Boström, P.; Tervahartiala, M.; Taimen, P.; Gleichenhagen, J.; Johnen, G.; Brüning, T.; Koch, S.; Roggisch, J.; Wirtz, R.M. Prognostic Role of Survivin and Macrophage Infiltration Quantified on Protein and mRNA Level in Molecular Subtypes Determined by RT-qPCR of KRT5, KRT20, and ERBB2 in Muscle-Invasive Bladder Cancer Treated by Adjuvant Chemotherapy. Int. J. Mol. Sci. 2020, 21, 7420.

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