Abstract 1329: Development of the first optimised “off the shelf” MAGE-A4 targeting TCR-NK cells for advancement into the clinic for the treatment of solid tumors

Abstract T cell receptor (TCR)-T based therapies, such as MAGE-A4 TCR-T cells, have shown compelling data demonstrating effective infiltration into and targeting of solid tumors with clinical responses across various solid cancers. However, heterogeneity and down-regulation of target antigen and HLA expression limit the durability and curative potential of these types of treatments. On the other hand, owing to their clinical potency, favorable safety profile and applicability as “off the shelf” therapy, natural killer (NK) cells have emerged as promising modalities in recent years. Unguided NK cells have shown limited potential against solid cancers, due to lacking infiltration into tumors. With our proprietary TCR-NK platform, we combine the solid tumor targeting capabilities of TCRs with the pan-cancer sensing and cytotoxic potential of highly potent killer cells, redirecting and arming NK cells with a fully functional TCR-CD3 complex. Here we present our lead product, an optimized TCR-NK cell product, ZI-MA4-1a, expressing an affinity enhanced TCR (KVL-a) directed to the HLA class I-restricted clinically validated cancer-testis antigen MAGE-A4, that is broadly expressed across solid tumors. The KVL-a TCR was successfully engineered from a naturally occurring wild-type TCR, increasing the affinity to MAGE-A4-peptide-HLA complex and resulting in enhanced recognition of tumor cells expressing the target antigen. ZI-MA4-1a is produced by transducing healthy peripheral blood derived NK cells with the full CD3 complex, the KVL-a TCR and a CD8 coreceptor. The process has been optimized to enrich TCR-NK cells, increasing process robustness, diminishing the impact of donor variability, and producing a higher performing TCR-NK cell product. We show the dual functionality of ZI-MA4-1a: TCR driven cytotoxicity as well as innate NK sensing of antigen knockout or HLA negative cancers. We demonstrate killing of heterogenous tumor cell models in vitro, as well as the multifunctionality of ZI-MA4-1a with killing, degranulation and multicytokine secretion. Interestingly, we observe enhanced proliferation of ZI-MA4-1a cells after activation by MAGE-A4 expressing tumor cells, indicating that TCR-NK cells acquire further proliferative capacity when the TCR is triggered. Finally, we demonstrate higher activity of ZI-MA4-1a compared to KVL-a and benchmark TCR transduced T cells against antigen positive targets, and activity against antigen negative tumor cells escaping TCR-T cell recognition. Citation Format: Sarah Vollmers, Artur Cieslar-Pobuda, Justyna Kmiecik, Giulia Malachin, Michelle L. Sætersmoen, Frida Høsøien Haugen, Margherita Boieri, Maja Sandve, Dennis Clement, Ines Cardoso, Pimthanya Wanichawan, Martha E. Haugstøyl, Anja Oldenburg, Liliane Christ, Pranav Oberoi, Luz Mora-Velandia, Anders Holm, Namir J. Hassan, Emilie Gauthy, Julia Ino, Sylvie Pollmann, Luise U. Weigand. Development of the first optimised “off the shelf” MAGE-A4 targeting TCR-NK cells for advancement into the clinic for the treatment of solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1329.