Digital pathology-aided assessment of tumor-infiltrating T lymphocytes in advanced stage, HPV-negative head and neck tumors

Emma J De Ruiter,Ruud H Brakenhoff,Stefan M Willems,Chris H J Terhaard,Remco De Bree,Reinout H De Roest,C René Leemans

doi:10.1007/s00262-020-02481-3

Abstract

AimThis study aimed to evaluate the presence and prognostic value of tumor-infiltrating T cells in the tumor epithelium in advanced stage, HPV-negative head and neck squamous cell carcinoma (HNSCC) patients treated with primary chemoradiotherapy using digital pathology.MethodsPre-treatment biopsies from 80 oropharyngeal, 52 hypopharyngeal, and 29 laryngeal cancer patients were collected in a tissue microarray (TMA) and immunohistochemically stained for T-cell markers CD3, CD4, CD8, FoxP3, and PD1, and for immune checkpoint PD-L1. For each marker, the number of positive tumor-infiltrating lymphocytes (TILs) per mm2 tumor epithelium was digitally quantified and correlated to overall survival (OS), disease-free survival (DFS), and locoregional control (LRC), as well as to clinicopathological characteristics. Differences in clinical outcome were estimated using Cox proportional hazard analysis and visualized using Kaplan–Meier curves.ResultsThe patient cohort had a 3-year OS of 58%, with a median follow-up of 53 months. None of the T-cell markers showed a correlation with OS, DFS or LRC. A low N stage was correlated to a better prognosis (OS: HR 0.39, p = 0.0028, DFS: HR 0.34, p = < 0.001, LRC: HR 0.24, p = 0.008). High TIL counts were more often observed in PD-L1-positive tumors (p < 0.05).ConclusionThis study showed an objective, digital pathology-aided method to assess TILs in the tumor epithelium. However, it did not provide evidence for a prognostic role of the presence of CD3 + , CD4 + , CD8 + , FoxP3 + , and PD1 + TILs in the tumor epithelium of advanced stage, HPV-negative HNSCC patients treated with primary chemoradiotherapy.

Highlights

Materials and methodsDespite the improvement of treatment outcome with the use of radiotherapy in combination with concomitant chemotherapy, an estimated 25–50% of all head and neck squamous cell carcinoma (HNSCC) patients still face locoregional recurrence and overall survival remains poor [1]
Almost all patients were treated with radiotherapy in combination with cisplatin
Five patients were treated with carboplatin instead of cisplatin. 24 patients were initially treated with cisplatin, but switched to carboplatin due to adverse events. 19 patients discontinued treatment after two doses of cisplatin, receiving a total dose of 200 mg/m2 body surface area

Summary

Introduction

Despite the improvement of treatment outcome with the use of radiotherapy in combination with concomitant chemotherapy, an estimated 25–50% of all head and neck squamous cell carcinoma (HNSCC) patients still face locoregional recurrence and overall survival remains poor [1]. Failure of locoregional control of HNSCC strongly contributes to morbidity and mortality [2, 3]. Identifying robust biomarkers predicting patients at risk for recurrent disease after therapy would be of great value in selecting the best treatment for each individual patient [4]. For many types of cancer, it has become clear that the interplay between tumor cells and their microenvironment strongly influences tumor aggressiveness and therapy resistance [5, 6]. Therapies targeting the anti-tumor immune response are rapidly evolving and are already implemented in a variety of cancer types [7]. In HNSCC, immune checkpoint inhibitors nivolumab and pembrolizumab are recently incorporated in clinical practice, with other immunotherapeutic agents probably soon to follow [8]

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