Hexokinase2-Engineered T Cells Display Increased Anti-Tumor Function

Raphaëlle Toledano Zur ¹ Shiran Didi Zurinam ¹ Maria Radman ¹ Elia Funaro Balouka ¹ Tatiana Borodianskiy-Shteinberg ¹ Dieter Saur ^2,3 Cyrille J Cohen ^1*

• ¹ The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat Gan, Israel
• ² German Cancer Research Center (DKFZ), Heidelberg, Baden-Württemberg, Germany
• ³ Technical University of Munich, Munich, Bavaria, Germany

T cells face significant metabolic challenges in the tumor microenvironment (TME), where cancer cells monopolize critical nutrients like glucose and amino acids. This metabolic competition supports tumor growth while impairing T-cell anti-tumor responses, partly by reducing glycolytic function. Hexokinase 2 (HK2), a key enzyme in glycolysis, plays a pivotal role in maintaining T-cell functionality.To enhance T-cell function, primary human T cells were genetically engineered to overexpress HK2 alongside a tumor-specific receptor. These engineered T cells were tested in vitro and in vivo to evaluate their metabolic and therapeutic efficacy.HK2-engineered T cells exhibited increased glycolytic capacity, leading to enhanced cytokine secretion, activation marker expression, and metabolic activity compared to controls. In vivo studies using a human tumor xenograft model demonstrated the superior therapeutic efficacy of HK2engineered T cells, including delayed tumor growth and improved survival.HK2 overexpression improves T-cell metabolic fitness and functionality in hostile TMEs, offering a promising foundation for the development of next-generation immunotherapies targeting T-cell metabolism.