Anti-PD-1 and Anti-CD39 Induce Respective CD8 T Cell and Macrophage Responses at the Patient-derived Ex Vivo Tumor-border

Le résumé fourni par la source

Abstract Rationale Efficacy of immunotherapies is critically dependent on the individual immune cell composition of the patient's tumor microenvironment. Presently, we aim to understand how the native microenvironment affects immune cell activation via immunotherapies using patient specific tumor slices. Methods We generated and immuno-characterized patient-derived precision cut lung slices from non-tumor (>10cm away from tumor-center) and tumor-border (part tumor and part tumor-adjacent tissue). These were treated with anti-CD3/CD28, anti-PD1 (50µM) or anti-CD39 (1µg/mL) for 30h. We leveraged flow cytometry, cytokine analysis, histology, bulk-RNAseq and viability assays to analyze these slices after treatment. Results Tumor-border slices are viable and contain higher frequencies of CD45+ (8.3-fold) and CD3+/CD8+ (6-fold) cells compared to non-tumor tissue. Additionally, tumor-border slices display increased secretion of IL-6 (30-fold), IL-2 (1.5-fold), and Granzyme B (6-fold) as well as increased RNA expression of TGFB1, IFNGR2 and MMP11, compared to non-tumor slices. Tumor-border CD8+ T cells highly expressed the activation markers CD137, CD107, PD-1, and CD39. Anti-CD3/CD28 treatment induced inflammatory T cell responses and pro-inflammatory tissue cytokine secretion in both non-tumor vs tumor-border slices evidenced by higher secretion of IFNγ (150-fold) and IL-2 (10-fold). Contrarily, anti-PD-1 treated tumor-border but not non-tumor slices secrete more IFNγ (1.6-fold) and perforin (1.5-fold) compared to the medium control. anti-PD-1 treatment also induced upregulation of the activation marker CD137 (1.8-fold) and degranulation marker CD107a (1.3-fold) on CD8+ T cells in the tumor-border. Furthermore, treatment with anti-CD39 induced upregulation of IL-1β (1.2-fold) in the tumor-border slices, indicating macrophage involvement. Indeed, there the MFI of CD163 was decreased in non-tumor macrophages and the MFI of HLA-DR and CD86 was higher on macrophages in tumor-border slices upon treatment with anti-CD39. Conclusion While anti-CD3/CD28 induces T cell responses in non-tumor and tumor-border slices, anti-PD-1 induces activation of T cells in the tumor microenvironment of tumor-border slices only, and in a donor-dependent manner. Additionally, anti-CD39 treatment induces a pro-inflammatory phenotype on macrophages in the tumor microenvironment. Finally, matched non-tumor and tumor-border slices provide a platform to understand the tissue inflammation both at baseline in the tumor-free tissue as well as in the tumor itself. Taken together, we demonstrated the added value of viable patient-derived tumor tissue, providing data on the efficacy of immunotherapies in context of patient specific tumor microenvironments.

Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.