Abstract 6716: DLL3 TriTCE Co-Stim: A next generation trispecific T cell engager with integrated CD28 costimulation for the treatment of DLL3-expressing cancers
Authors: Repenning Peter , Lau Desmond , Hernaez Diana C. , Robinson Alec , Escanda Diego P. , Rocha Mariana , Moreno Begonia S. , Zhang John , Shao Polly , Escalante Nichole , Newhook Lisa , Bhojane Purva , Piscitelli Chayne L. , Moore Paul A. , Von Kreudenstein Thomas Spreter , Weisser Nina E. 
Published: March 2024

Abstract
Small cell lung cancer (SCLC) is an aggressive neuroendocrine cancer with a poor prognosis and high unmet medical need. Current standards of care, including chemotherapy, targeted therapy and immunotherapy, are associated with limited duration of response. Furthermore, SCLC is characterized by an immunosuppressive microenvironment and poor T cell infiltration which present additional treatment challenges. Delta-like ligand 3 (DLL3) is an inhibitory Notch ligand that is aberrantly overexpressed in SCLC and other neuroendocrine tumors, but minimally expressed in normal tissues, and is a new therapeutic target. Bispecific T cell engagers (TCE) targeting DLL3 have entered the clinic and demonstrated promising anti-tumor activity in SCLC patients; however, we believe there is room to improve on the rate and depth of response. To address treatment challenges and enhance the durability and sustainability of T cell activation, we have engineered a trispecific co-stimulatory T cell engager (TriTCE Co-stim) molecule that optimally engages CD3 and CD28 and redirects and enhances cytotoxic T cell responses to DLL3-expressing tumor cells while maintaining a desired safety profile. A panel of DLL3xCD3xCD28 trispecific antibodies (Abs) was engineered with a conventional anti-CD28 agonist paratope in a variety of Ab formats, geometries, paratope affinities and specificities. Geometry and design features of the tri-specific Abs were informed by prior TriTCE Co-stim molecule optimization1. AzymetricTM and EFECTTM platforms were used to facilitate heterodimeric multi-chain antibody assembly with silenced Fc gamma function. Lead TriTCE Co-stim Abs were selected for enhanced DLL3-dependent antitumor cytotoxic activity and increased T cell function including T cell activation, proliferation and cytokine production, compared to competitor bispecific TCEs. DLL3-independent activity and cytokine release were assessed with human PBMCs in solid and solution phase assays to evaluate the potential for cytokine release induction. In summary, we have identified multiple TriTCE Co-Stim Ab formats with improved DLL3-dependent cytotoxicity and T cell activation over competitor bispecific TCEs across multiple DLL3-expressing tumors. By screening various Ab formats, geometries and paratope affinities, we have selected lead DLL3 TriTCE Co-stim Abs displaying target-dependent activation of T cells with potent anti-tumor activity which may translate to improved and more durable antitumor responses in the treatment DLL3-expressing cancers.
Citation Format: Peter Repenning, Desmond Lau, Diana C. Hernaez, Alec Robinson, Diego P. Escanda, Mariana Rocha, Begonia S. Moreno, John Zhang, Polly Shao, Nichole Escalante, Lisa Newhook, Purva Bhojane, Chayne L. Piscitelli, Paul A. Moore, Thomas Spreter Von Kreudenstein, Nina E. Weisser. DLL3 TriTCE Co-Stim: A next generation trispecific T cell engager with integrated CD28 costimulation for the treatment of DLL3-expressing cancers [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 6716.