Tumor-Infiltrating T Cells Can Be Expanded Successfully from Primary Uveal Melanoma after Separation from Their Tumor Environment

Gülçin Gezgin; Marten Visser; Dina Ruano; Saskia J Santegoets; Noel F C C de Miranda; Pieter A van der Velden; Gregorius P M Luyten; Sjoerd H van der Burg; Els M Verdegaal; Martine J Jager

doi:10.1016/j.xops.2022.100132

Tumor-Infiltrating T Cells Can Be Expanded Successfully from Primary Uveal Melanoma after Separation from Their Tumor Environment

Ophthalmol Sci. 2022 Mar 1;2(2):100132. doi: 10.1016/j.xops.2022.100132. eCollection 2022 Jun.

Affiliations

¹ Department of Ophthalmology, Leiden University Medical Center, Leiden, The Netherlands.
² Department of Medical Oncology, Oncode Institute, Leiden University Medical Center, Leiden, The Netherlands.
³ Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.

Abstract

Purpose: To evaluate whether expanded tumor-infiltrating lymphocytes (TILs) can be obtained from primary uveal melanoma (UM) for potential use as adjuvant treatment in patients at risk of developing metastatic disease.

Design: Experimental research study.

Participants: Freshly obtained primary UM from 30 patients.

Methods: Three different methods were used to expand TILs: (1) direct culture from small fragments of fresh tumor tissue, (2) single-cell tissue preparation by enzymatic digestion and subsequent enrichment of mononuclear cells, and (3) selection of CD3⁺ T cells using magnetic beads. Surface expression of costimulatory and inhibitory T-cell markers and T-cell reactivity against autologous tumor cells was assessed. Clinical, histopathologic, genetic, and immunologic characteristics of the tumors were compared with the capacity to expand TILs and with their reactivity against autologous tumor cells.

Main outcome measures: The feasibility of expanding TILs from primary UM, testing their reactivity to autologous UM cells, and evaluating the impact of an immunomodulatory environment.

Results: Direct culture of tumor parts led to successful TIL culture in 4 of 22 tumors (18%), enrichment of mononuclear cells gave rise to TILs in 5 of 12 tumors (42%), while preselection of CD3⁺ T cells with magnetic beads resulted in TIL expansion in 17 of 25 tumors (68%). In 8 of 17 tumors (47%), the TIL cultures comprised UM-reactive T cells. The presence of UM-reactive T cells among TILs was not related to clinical, histologic, genetic, or immunological tumor characteristics. Interestingly, RNA-Seq analysis showed that approximately half of the UM tumors displayed an increased expression of immunomodulatory molecules related to T-cell suppression, such as galectin 3, programmed death-ligand 1, cytotoxic T-lymphocyte-associated protein 4, indoleamine 2,3-dioxygenase 1, and lymphocyte activating 3, potentially explaining why T cells require optimal removal of tumor components for expansion.

Conclusions: The need to separate TILs from their tumor microenvironment for their successful expansion and the presence of UM-reactive T cells among TILs suggests that these UM-reactive T cells are strongly suppressed in vivo and that UM is immunogenic. These findings indicate that adoptive TIL therapy could be an option as an adjuvant treatment in primary UM patients at high risk of developing metastatic disease.

Keywords: BAP1, BRCA1-associated protein 1; CTLA4, cytotoxic T-lymphocyte-associated protein 4; IFN-γ, interferon γ; IL, interleukin; Immunotherapy; PD-1, programmed cell death protein 1; TILs, tumor-infiltrating lymphocytes; TIM-3, T-cell immunoglobulin and mucin-domain containing 3; TME, tumor microenvironment; Tumor-infiltrating T cells; Tumor-infiltrating lymphocytes; UM, uveal melanoma; Uveal melanoma.