Het inzetten van Vγ9Vδ2-T cellen als immunotherapie voor kanker

In this thesis, we explored the efficacy of Vγ9Vδ2-T cell-targeting bsTCEs in inducing anti-tumor activity in a solid cancer setting, and determined the behavior of Vγ9Vδ2-T cells in an organotypic human melanoma-in-skin model. In chapter 2, we present the generation and functional testing of a bsTCE targeting the Vδ2-TCR and epidermal growth factor receptor (EGFR). This bsTCE demonstrated anti-tumor efficacy in in vitro, ex vivo, and in vivo experiments which was accompanied by Vγ9Vδ2-T cell activation, as well as bystander activation of conventional T and NK cells through TNF and IFNγ production. Cytotoxicity of normal (EGFR expressing) tissue was not observed. We additionally explored the safety profile of targeting Vγ9Vδ2-T cells to EGFR in non-human primates (NHP) and found no clinical, biochemical, or histopathologic signs of toxicity despite target engagement. These promising pre-clinical results formed the foundation for an ongoing phase 1 clinical study (NCT05983133) evaluating the safety and tolerability of PF-08046052 (LAVA-1223) in patients with advanced (EGFR-expressing) solid tumors. In chapter 3, we describe the pre-clinical development of a bsTCE targeting the Vδ2-TCR and prostate-specific membrane antigen (PSMA) as novel investigational prostate cancer treatment. This bsTCE mediated lysis of multiple PSMA-expressing prostate tumors without affecting non-malignant prostate tissue. NKG2D and DNAM-1 receptors contributed to Vγ9Vδ2-T cell activation and tumor lysis at low concentrations of PSMA-Vδ2 bsTCEs. In a mouse model, the bsTCE inhibited tumor growth, and in vitro experiments revealed its capacity to induce Vγ9Vδ2-T cell mediated antigen cross-presentation to antigen-specific CD8+ T cells. Tissue-cross reactivity studies confirmed the specificity of binding to tissues expressing PSMA or containing Vγ9Vδ2-T cells. These findings served as the foundation for the phase 1/2a clinical study of LAVA-1207 in patients with therapy-refractory metastatic castration-resistant prostate cancer (NCT05369000). Chapter 4 delves into the anti-tumor efficacy of a Vδ2 and PD-L1-targeting bsTCE with additional PD-L1/ PD-1 blocking capacity. BsTCE-activated Vγ9Vδ2-T cells induced lysis of both cell line-derived and cancer patient-derived PD-L1 expressing tumor cells, as well as three-dimensional (3D) melanoma tumor spheroids. The bsTCE not only increased the infiltration of Vγ9Vδ2-T cells into 3D tumor spheroids but also enhanced their activation status. The addition of the PD-L1 bsTCE and Vγ9Vδ2-T cells resulted in elevated levels of activated CD4+ and CD8+ T cells, along with more mature dendritic cells and reduced numbers of PD-L1+ macrophage-like cells in cultures of patient derived melanoma and renal cell carcinoma suspensions. The PD-L1 bsTCE addresses key challenges of current therapies by simultaneously targeting PD-L1-expressing tumor and myeloid cells while also activating conventional T cells. We describe in chapter 5 the characterization of different multi-valent Vγ9Vδ2-TCR engagers to induce expansion and activation of effector Vγ9Vδ2-T cells to enhance anti-tumor responses. These multi-valent engagers induced expansion of Vγ9Vδ2-T cells in vitro and in an in vivo model in NOG-hIL-15 mice. Expanded Vγ9Vδ2-T cells had an effector memory phenotype with high expression levels of activation markers combined with enhanced anti-tumor effector functions. Chapter 6 examines the immunotherapeutic potential of Vγ9Vδ2-T cells within the context of a Mel-RhS model. In this model, Vγ9Vδ2-T cells were found to be in close proximity to or within tumor nests, with a fraction being decorated by melanoma-associated chondroitin sulfate proteoglycan (MCSP). MCSP displaying Vγ9Vδ2-T cells expressed higher levels of 4-1BB, NKp44, PD-1, and PD-L1 than their MCSP- counterparts. These observations underscore the ability of Vγ9Vδ2-T cells to actively navigate the TME and target and trogocytose cancer cells. In chapter 7, we will summarize and discuss our findings described in this thesis in the context of recent and anticipated future developments in the field.