The mutational load and a T-cell inflamed tumour phenotype identify ovarian cancer patients rendering tumour-reactive T cells from PD-1 + tumour-infiltrating lymphocytes
Diego Salas-Benito 1 2, Enrique Conde 2 3, Ibon Tamayo-Uria 2 3, Uxua Mancheño 2 3, Edurne Elizalde 2 3, David Garcia-Ros 2 4 5, Jose M Aramendia 1 2, Juan C Muruzabal 2 6, Julia Alcaide 7, Francisco Guillen-Grima 2 8, Jose A Minguez 1 9, Jose Amores-Tirado 10, Antonio Gonzalez-Martin 1 11, Pablo Sarobe 2 3 12, Juan J Lasarte 2 3, Mariano Ponz-Sarvise 13 14 15, Carlos E De Andrea # 2 4 5 16, Sandra Hervas-Stubbs # 17 18 19
Background: Adoptive immunotherapy with tumour-infiltrating lymphocytes (TIL) may benefit from the use of selective markers, such as PD-1, for tumour-specific T-cell enrichment, and the identification of predictive factors that help identify those patients capable of rendering tumour-reactive TILs. We have investigated this in ovarian cancer (OC) patients as candidates for TIL therapy implementation.
Methods: PD-1- and PD-1+ CD8 TILs were isolated from ovarian tumours and expanded cells were tested against autologous tumour cells. Baseline tumour samples were examined using flow cytometry, multiplexed immunofluorescence and Nanostring technology, for gene expression analyses, as well as a next-generation sequencing gene panel, for tumour mutational burden (TMB) calculation.
Results: Tumour-reactive TILs were detected in half of patients and were exclusively present in cells derived from the PD-1+ fraction. Importantly, a high TIL density in the fresh tumour, the presence of CD137+ cells within the PD-1+CD8+ TIL subset and their location in the tumour epithelium, together with a baseline T-cell-inflamed genetic signature and/or a high TMB, are features that identify patients rendering tumour-reactive TIL products.
Conclusion: We have demonstrated that PD-1 identifies ovarian tumour-specific CD8 TILs and has uncovered predictive factors that identify OC patients who are likely to render tumour-specific cells from PD-1+ TILs.