Genetic modification of NK cells with anti-CD19 CAR expressing GITRL to increase NK cells effector function for leukemias and lymphomas treatment

Abstract

The development of chimeric antigen receptor T cells (CAR-T) therapy brought significant advances in hematological cancers treatment. It combines the cytotoxic activity of autologous T cells with antibody specificity. For refractory or relapsed B-cell malignancies, the CD19 antigen has been the main target explored, as it is expressed mainly in B cells. Despite an increase in remission rates with anti-CD19 treatment, the application of T cells is expensive and associated with severe adverse effects. An alternative is the use of NK cells, which do not offer the potential to cause graft versus host disease, allowing allogeneic cells use for an off-the-shelf application. NK cells can also act in tumor cells elimination through intrinsic cytotoxic mechanisms. However, optimizations are needed for CAR-NK cells production. This project aims to establish the production of anti-CD19 allogeneic CAR-NK cells with a novel construct that includes GITRL molecule. NK-92 cell line, peripheral blood and cord blood NK cells will be transduced with a lentiviral vector containing CAR sequence. Methods for efficiently cryopreserve generated cells will be tested. The production of cytokines will be evaluated in vitro as well as its cytotoxic activity against tumor B cells. Furthermore, cells persistence and its antineoplasic potential will be evaluated in vivo using a Burkitt lymphoma model. The development of this anti-CD19 CAR-NK cells with a novel construct may lead to the development of an efficient and more affordable off-the-shelf therapy.