Project phases and realisation

Our research project aims to fill the gaps described in the “Short description” section to improve CAR T cells targeting CD19-positive hematological malignancies. We will achieve this by:

(i) Incorporating the key transcription factors (TFs) identified in our previous studies as important candidates for improving the intrinsic T cell properties; and

(ii) Genetic integration of these candidate molecules into CAR T cells to enable constitutive or inducible expression (TF-CAR T cells).

In Objective 1, we will utilize a novel genetic platform called Uni-Vect, developed by the PI of this project, while he was working at the University of Pennsylvania (UPenn). Uni-Vect will allow for transient expression of TFs in CAR T cells. Additionally, we will design and test systems with constitutively expressed TFs (cTF-CAR T cells). We will create CD19-targeting CAR T cells upgraded with ectopic expression of two key transcription factors with a hypothesis to create CAR T cells with a fitness profile associated with a capacity to expand, persist and mediate cancer regression after infusion.

In Objective 2, we will establish in vitro systems that will enable us to evaluate and compare functional and phenotypic properties of i/cTF-CAR T cells. Using advanced immunological approaches, we will examine the mechanisms that lead to the potential improved functions. In collaboration with our partners from the US, we will also investigate whether i/cTF-CAR T cells can improve activity against chronic lymphocytic leukemia (CLL) and improve T cell manufacturing in difficult-to-expand T cells.

In Objective 3, we will test i/cTF-CAR T cell products in xenograft and syngeneic preclinical mouse models and determine the impact of ectopic expression of TFs on CAR T cell expansion, persistence and anti-tumor activity in vivo. We will isolate CAR T cells from mice and examine CAR T cell phenotype and function to gain a better understanding of which key factors contribute to improved T-cell intrinsic properties. This will inform further developments of i/cTF-CAR T cell approaches.