J Cancer 2018; 9(12):2191-2202. doi:10.7150/jca.22846

Research Paper

Technical Feasibility of Tissue Microarray (TMA) Analysis of Tumor-Associated Immune Response in Prostate Cancer

Timothy J. Wallace1, Junqi Qian2, Itzhak Avital3, Curt Bay4, Yan-Gao Man5, Laurie L. Wellman6, Chris Moskaluk7, Dean Troyer6, Dharam Ramnani2, Alexander Stojadinovic8✉

1. Bon Secours Cancer Institute, Richmond, Virginia, U.S.A.
2. Virginia Urology, Richmond, Virginia, U.S.A.
3. Soroka University Center for Advanced Cancer Care, Ber Sheva, Israel
4. A.T. Still University, Mesa, Arizona, U.S.A.
5. National Medical Centre of Colorectal Disease, Third Affiliated Hospital of Nanjing University of Traditional Chinese Medicine (TCM), Nanjing, China
6. Eastern Virginia Medical School, Norfolk, Virginia, U.S.A.
7. University of Virginia, Charlottesville, Virginia, U.S.A.
8. Uniformed Services University of the Health Sciences, Bethesda, Maryland

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license (https://creativecommons.org/licenses/by-nc/4.0/). See http://ivyspring.com/terms for full terms and conditions.
Wallace TJ, Qian J, Avital I, Bay C, Man YG, Wellman LL, Moskaluk C, Troyer D, Ramnani D, Stojadinovic A. Technical Feasibility of Tissue Microarray (TMA) Analysis of Tumor-Associated Immune Response in Prostate Cancer. J Cancer 2018; 9(12):2191-2202. doi:10.7150/jca.22846. Available from http://www.jcancer.org/v09p2191.htm

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Introduction: The androgen receptor (AR) regulates immune-related epithelial-to-mesenchymal transition (EMT), and prostate cancer (PCa) metastasis. Primary tumor-infiltrating lymphocytes (TILs) [CD3+, CD4+, and CD8+ TILs] are potential prognostic indicators in PCa, and variations may contribute to racial disparities in tumor biology and PCa outcomes.

Aim: To assess the technical feasibility of tumor microarray (TMA)-based methods to perform multi-marker TIL profiling in primary resected PCa.

Methods: Paraffin-embedded tissue cores of histopathologically-confirmed primary PCa (n = 40; 1 TMA tissue specimen loss) were arrayed in triplicate on TMAs. Expression profiles of AR, CD3+, CD4+, and CD8+ TILs in normal prostate, and the center and periphery of both the tumor-dominant nodule and highest Gleason grade were detected by IHC and associated with clinical and pathological data using standard statistical methodology. An independent pathologist, blinded to the clinical data, scored all samples (percent and intensity of positive cells).

Results: TMAs were constructed from 21 (53.8%) Black and 18 (46.2%) White males with completely-resected, primarily pT2 stage PCa [pT2a (n = 3; 7.7%); pT2b (n = 2; 5.1%); pT2c (n = 27; 69.2%); pT3a (n = 5; 12.8%); mean pre-op PSA = 8.17 ng/ml]. The CD3, CD4, CD8, and CD8/CD3 cellular protein expression differed from normal in the periphery of the dominant nodule, the center of the highest Gleason grade, and the periphery of the highest Gleason grade (P < 0.05). Correlations between TIL expression in the center and periphery of the dominant nodule, with corresponding center and periphery of the highest Gleason grade, respectively, were robust, and the magnitude of these correlations differed markedly by race (P < 0.05).

Conclusions: Multi-marker (AR, CD3, CD4, CD8) profiling with IHC analysis of TMAs consisting of primary, non-metastatic resected prostate cancer is technically feasible in this pilot study. Future studies will evaluate primary tumor immunoscore using semi-quantitative, IHC-based methodology to assess differences in the spectrum, quantity, and/or localization of TILs, and to gain insights into racial disparities in PCa tumor biology and clinical outcomes.

Keywords: Prostate cancer, Immunoscore, Tissue microarray, Tumor-infiltrating lymphocytes, Androgen receptor