Abstract 2829: Non-viral precision genome engineering enables personalized adoptive neoTCR T cell therapy for cancer including multiple additional edits that improve the activity of neoTCR T cells by enhancing CD4 T cell antigen sensitivity and conferring resistance to TGFβ
Authors: Tran Charles W. , Lee Kayla , Purandare Bhamini , Byers James , Dubreuil Michael M. , Lu William , Mass Michal , Jacoby Kyle , Mandl Stefanie J. , Sennino Barbara 
Published: June 2022

Personalized autologous TCR-T cell therapies targeting neoepitopes (neoE) derived from tumor-specific mutations are a compelling approach for the treatment of patients with solid tumors. Using the ultra-sensitive imPACT Isolation Technology®, antigen-experienced, neoE-specific CD8 T cells are captured from the blood of patients with solid cancers followed by cloning of the cognate, neoE-specific, MHC class I-restricted T cell receptors (neoTCRs). DNA-mediated (non-viral) precision genome engineering technology is then used to engineer autologous CD8 and CD4 T cells to express the neoTCR. We have further built upon this platform to modify neoTCR-T cells to address potential sources for tumor immune evasion.

The versatility of this single-step gene editing platform is demonstrated here by the presentation of a neoTCR T cell product with knockout of endogenous TCRα and TCRβ and the simultaneous expression of a neoTCR with CD8 co-receptor (CD8coR) from the same expression cassette along with knockout of TGFβ receptor 2 (TGFBR2). We have previously shown that CD8coR expression augments the activity of CD4 T cells engineered to express MHC class I restricted neoTCRs, by increasing CD4 neoTCR T cell helper and effector function. TGFβ is known to promote tumor growth, metastasis, and epithelial to mesenchymal transition (EMT) and to inhibit effector immune responses while promoting fibrosis and the differentiation of inhibitory cell types such as regulatory T cells. TGFβ is expressed by many tumor types and high TGFβ expression is associated with worse prognosis for several cancer subtypes including colorectal, lung and glioblastoma. TGFBR2 is a critical mediator of TGFβ signaling, resulting in potent, inhibitory effects on T cell function.

Our data show that simultaneous ablation of TGFBR2 signaling and expression of CD8coR was achieved with high efficiency, resulting in fully functional CD8 and CD4 neoTCR-T cells. Deletion of TGFBR2 with or without CD8coR expression, together with the expression of a neoE-targeted TCR preserved T cell effector function in the presence of inhibitory concentrations of TGFβ. Importantly, the combination of TGFBR2 knockout and CD8coR expression resulted in additive benefits, providing proof-of-concept for modifying two orthogonal features to improve neoTCR T cell function. Altogether, these results demonstrate the applicability of this versatile, precision genome engineering platform technology to yield enhanced, next-generation neoTCR-T cell therapies to expand the potential for clinical benefit in persons with solid cancers.

Citation Format: Charles W. Tran, Kayla Lee, Bhamini Purandare, James Byers, Michael M. Dubreuil, William Lu, Michal Mass, Kyle Jacoby, Stefanie J. Mandl, Barbara Sennino. Non-viral precision genome engineering enables personalized adoptive neoTCR T cell therapy for cancer including multiple additional edits that improve the activity of neoTCR T cells by enhancing CD4 T cell antigen sensitivity and conferring resistance to TGFβ [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2829.