Abstract 3478: Pre-clinical characterization of AZD0240, a CRISPR-engineered autologous TCR-T cell product targeting KRAS-G12D/HLA-A*11:01 that is multi-armored to increase epitope sensitivity

KRAS is one of the most commonly mutated genes in human cancer, with the G12D mutation having the highest frequency, occurring in 33% and 28% of colorectal cancer (CRC) and pancreatic cancer (PDAC), respectively. KRAS G12D-containing peptides are presented by the common HLA allele HLA-A*11:01, making this cancer neoantigen a highly attractive target for the development of adoptive TCR-T cell therapy. However, the levels of cancer neoantigen presentation are typically low and combined with the immunosuppressive tumor microenvironment this compromises the activation and reactivity of TCR-engineered T cells, posing a major hurdle for these cells to eradicate tumor lesions. To establish a TCR-T cell therapy clinically intended for the treatment of solid cancers, including CRC and PDAC, we developed AZD0240, a CRISPR-engineered autologous TCR-T cell product targeting KRAS-G12D/HLA-A*11:01. To improve the reactivity of AZD0240 to the KRAS-G12D neoantigen, we developed a novel multi-armoring strategy, involving the combined disruption of a key negative regulator of T cell function and the introduction of a positive enhancement of TCR signaling. Employing this multi-armoring strategy, we observed enhanced cytokine production in responds to low levels of exogenously loaded KRAS-G12D peptide, as well as HLA-A*11:01+ tumor cells that express the KRAS-G12D mutation. Moreover, assessment of the in vitro and in vivo proliferative and cytotoxic capacity revealed that the AZD0240 armoring strategy can profoundly increase both the proliferative capacity and the anti-tumor efficacy of TCR edited T cells. Notably, the armored AZD0240 T cells were unable to respond to the KRAS G12 wild-type epitope, and no signs of increased TCR cross- or allo-reactivity, or malignant transformation were detected, derisking the clinical use of this armoring strategy. Taken together, AZD0240 is a KRAS-G12D/HLA-A*11:01 targeted TCR-T cell therapy that employs a novel multi-armoring strategy and is currently in preparation for clinical testing, with encouraging signs of pre-clinical efficacy and safety. Citation Format: Lianne Kok, Sander Eshuis, Xiangjun Kong, Jennifer Ma, Michelle Mojadidi, Paula Kroon, Vanessa Tubb, Ton Schumacher, Arianne Perez, Carsten Linnemann, Mark Cobbold, Jeroen van Heijst, Gavin Bendle. Pre-clinical characterization of AZD0240, a CRISPR-engineered autologous TCR-T cell product targeting KRAS-G12D/HLA-A*11:01 that is multi-armored to increase epitope sensitivity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3478.