Immunohistochemical analyses of paraffin-embedded sections after primary surgery or trimodality treatment in esophageal carcinoma

Benjamin Terfa Igbo; Annett Linge; Susanne Frosch; Theresa Suckert; Liane Stolz-Kieslich; Steffen Löck; Mani Sankari Kumaravadivel; Thilo Welsch; Jürgen Weitz; Ulrich Sommer; Daniela Aust; Esther G C Troost

doi:10.1016/j.ctro.2022.08.001

Immunohistochemical analyses of paraffin-embedded sections after primary surgery or trimodality treatment in esophageal carcinoma

Clin Transl Radiat Oncol. 2022 Aug 3:36:106-112. doi: 10.1016/j.ctro.2022.08.001. eCollection 2022 Sep.

Authors

Benjamin Terfa Igbo^{1

2}, Annett Linge^{2

3

4

5}, Susanne Frosch^{3

5}, Theresa Suckert^{2

4}, Liane Stolz-Kieslich⁴, Steffen Löck^{1

2

3}, Mani Sankari Kumaravadivel^{2

4}, Thilo Welsch⁶, Jürgen Weitz^{6

5}, Ulrich Sommer⁷, Daniela Aust^{7

8}, Esther G C Troost^{1

2

3

4

5}

Affiliations

¹ Institute of Radiooncology - OncoRay, Helmholtz-Zentrum Dresden-Rossendorf, Rossendorf, Germany.
² OncoRay - National Center for Radiation Research in Oncology, Faculty of Medicine and University Hospital Carl Gustav Carus, Technische Universität Dresden, Helmholtz-Zentrum Dresden - Rossendorf, Dresden, Germany.
³ Department of Radiotherapy and Radiation Oncology, Faculty of Medicine and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
⁴ German Cancer Consortium (DKTK), Partner Site Dresden, and German Cancer Research Center (DKFZ), Heidelberg, Germany.
⁵ National Center for Tumor Diseases (NCT), Partner Site Dresden, Germany; German Cancer Research Center (DKFZ), Heidelberg, Germany; Faculty of Medicine and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany, and; Helmholtz Association / Helmholtz-Zentrum Dresden - Rossendorf (HZDR), Dresden, Germany.
⁶ Department of Visceral, Thoracic and Vascular Surgery (VTG), Faculty of Medicine and University Hospital Carl Gustav Carus, Dresden, Germany.
⁷ Institute of Pathology, Faculty of Medicine and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany.
⁸ Institute for Pathology and Tumor and Normal Tissue Bank of the University Cancer Center (UCC), University Hospital Carl Gustav Carus, Medical Faculty, Technische Universität Dresden, Dresden, Germany.

Abstract

Background: The microscopic tumor extension before, during or after radiochemotherapy (RCHT) and its correlation with the tumor microenvironment (TME) are presently unknown. This information is, however, crucial in the era of image-guided, adaptive high-precision photon or particle therapy.

Materials and methods: In this pilot study, we analyzed formalin-fixed paraffin-embedded (FFPE) tumor resection specimen from patients with histologically confirmed squamous cell carcinoma (SCC; n = 10) or adenocarcinoma (A; n = 10) of the esophagus, having undergone neoadjuvant radiochemotherapy followed by resection (NRCHT + R) or resection (R)]. FFPE tissue sections were analyzed by immunohistochemistry regarding tumor hypoxia (HIF-1α), proliferation (Ki67), immune status (PD1), cancer cell stemness (CXCR4), and p53 mutation status. Marker expression in HIF-1α subvolumes was part of a sub-analysis. Statistical analyses were performed using one-sided Mann-Whitney tests and Bland-Altman analysis.

Results: In both SCC and AC patients, the overall percentages of positive tumor cells among the five TME markers, namely HIF-1α, Ki67, p53, CXCR4 and PD1 after NRCHT were lower than in the R cohort. However, only PD1 in SCC and Ki67 in AC showed significant association (Ki67: p = 0.03, PD1: p = 0.02). In the sub-analysis of hypoxic subvolumes among the AC patients, the percentage of positive tumor cells within hypoxic regions were statistically significantly lower in the NRCHT than in the R cohort across all the markers except for PD1.

Conclusion: In this pilot study, we showed changes in the TME induced by NRCHT in both SCC and AC. These findings will be correlated with microscopic tumor extension measurements in a subsequent cohort of patients.

Keywords: 5-FU, 5-Fluorouracil; AC, Adenocarcinoma; AUC, Area under curve; BSA, Body surface area; CT, Computed tomography; CTV, Clinical target volume; CXCR4, Chemokine receptor type 4; Esophageal cancer; FDG, [18F]-fluorodeoxyglucose; FFPE, Formalin-fixed paraffin-embedded; GTV, Gross tumor volume; HIF-1α, Hypoxia-inducible factor 1-alpha; HNSCC, Head and neck squamous cell carcinoma; IgG, Immunoglobulin; Ki67, Tumor proliferation nuclear protein; MRI, Magnetic resonance imaging; Microscopic tumor extension; NRCHT +R, Neoadjuvant radiochemotherapy followed by resection; PD1, Programmed death 1 receptor; PET, Positron emission tomography; PTV, Planning target volume; R, Resection; RCHT, Radiochemotherapy; Radiochemotherapy; SCC, Squamous cell carcinoma; TME, Tumor microenvironment; Tumor microenvironment; UKD, University Hospital Carl Gustav Carus Dresden; Whole slide image analysis; p53, Tumor suppressor protein.