The role of oxidative stress in adoptive therapy with the use of chimeric antigen receptors

The aim of this project is to elucidate the mechanisms regulating the functions of cytotoxic cells in oxidative tumor microenvironment (TME) and to identify strategies potentiating the efficacy of chimeric antigen receptor (CAR)-based adoptive cell therapy in preclinical models. TME is characterized by an increased inflammation that is associated with the enhanced production of reactive oxygen species (ROS) which in turn generate oxidative stress that impairs antitumor response. Our preliminary results indicate that individual subsets of lymphocytes have different redox capacity. Moreover, we discovered that the construction of CARs used to modify effector cells influence their viability and ability to exert cytotoxic functions in prescence of oxidative stress. Thus, within this project, we will specifically determine the antioxidant potential and ability of various populations of cytotoxic cells to kill target tumor cells in the presence of elevated ROS levels. Next, we will explore the influence of various co-stimulating domains on antioxidant potential of CAR-modified effector cells. To further improve the cytotoxic functions of CAR-modified cells in highly oxidative milieu we will modulate the expression of selected antioxidant enzymes by genetic modifications. Finally, we will evaluate the efficacy of oxidant-resistant CAR-bearing cytotoxic cells both in vitro and in vivo.