Poster Presentation Melbourne Immunotherapy Network Winter Symposium 2021

CRISPR-Mediated PTPN2 Deletion in CAR T Cells Enhances Anti-Tumour Efficacy (#107)

Xin Du 1 2 , Florian Wiede 1 3 4 , Phil Darcy 1 2 , Tony Tiganis 1 3 4
  1. Peter MacCallum Cancer Centre, Melbourne, VIC, Australia
  2. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Melbourne, VIC, Australia
  3. Department of Biochemistry and Molecular Biology, Monash University, Melbourne, VIC, Australia
  4. Monash Biomedicine Discovery Institute, Monash University, Melbourne, VIC, Australia

Chimeric Antigen Receptor T cell (CAR T) immunotherapy has been remarkably successful in the treatment of B-Cell Acute Lymphoblastic Leukaemia. However, beyond haematological malignancies, CAR T cell therapy has been ineffective in treating solid tumours. Novel approaches for enhancing the ability of CAR T cells to combat solid tumours are urgently required. Protein tyrosine phosphatases (PTPs) are enzymes that regulate a wide range of physiological processes including metabolism, cellular growth, proliferation and differentiation by controlling tyrosine phosphorylation-dependent signalling. PTPs are key regulators of T cell signalling and contribute to the maintenance of immune tolerance. Studies from our group have shown that PTPN2 plays a pivotal role in negatively regulating T cell receptor (TCR) signalling by dephosphorylating and inactivating the Src family protein tyrosine kinase LCK (Wiede, Shields et al. 2011). PTPN2 also attenuates cytokine signalling by dephosphorylating JAK-1 and JAK-3 and their target substrates STAT-1, -3 and -5 in a cell context-dependent manner. Since CARs signal via LCK, and cytokine signalling is critical for CAR T cell function, we postulated that inhibiting PTPN2 might bolster the anti-tumour activity of CAR T cells. Here we used CRISPR-Cas9-ribonucleoprotein (RNP)-mediated genome editing to delete PTPN2 in CAR T cells. Using this approach PTPN2 was efficiently deleted in CAR T cells and the deletion of PTPN2 significantly enhanced the anti-tumour efficacy CAR T cells in vitro and in vivo.