Type: Oral
Session: 702. CAR-T Cell Therapies: Basic and Translational: Novel Targets and Therapeutic Approaches for CAR-T Cells
Hematology Disease Topics & Pathways:
Research, Translational Research, Immunology, Metabolism, Biological Processes
To investigate the biological impact of IDH1 R132H neomorph overexpression (IDH1NM OE) in CAR T cells, we developed a bicistronic expression system for CD19-specific 2nd generation CAR with 4-1BB/CD3ζ to co-express IDH1 R132H neomorph (IDH1NM) and used wildtype IDH1 (IDH1WT) as a control. Western blotting confirmed IDH1WT and IDH1NM overexpression in manufactured CAR19 T cells. IDH1NM OE led to significant D-2HG accumulation in CAR19 T cells, unlike IDH1WT. The expression of IDH1NM did not affect the cytolytic activity of CAR T cells, confirming the preservation of CAR their effector function. IDH1NM OE, however, significantly augmented CAR T cell response in a repeat tumor challenge model, similar to our previously published TET2 knock-down studies in normal donor T cells (Fraietta et al., Nature 2018). Further, IDH1NM CAR T cells showed significantly increased outgrowth in the CCR7+CD45RA+ central memory phenotype during chronic CAR stimulation. Additionally, we observed that effector cytokine levels, including IFNɣ and TNFɑ, were significantly reduced along with a decrease in CD107a, a major degranulation marker, while IL-2 levels were increased which is consistent with our TET2 knock-down studies. This suggests that IDH1NM OE improves the maintenance of early memory as well as effector function in the face of repeated tumor cell encounter.
Next, we aimed to identify the relevance between metabolic reprogramming and memory function mediated by IDH1NM OE in CAR T cells, based on our hypothesis that IDH1NM OE impact ɑKG-dependent dioxygenase activity including various histone demethylase as well as TET2 in CAR T cells. We observed an increase in methylated histone and a decrease in oxidated cytosine in IDH1NM CAR T cells which experienced repeated CAR stimulation in vitro. This suggests that metabolic reprogramming by IDH1NM OE in CAR T cells leads to epigenetic alterations toward a memory-like phenotype in CAR T cells. Our studies therefore confirm that the metabolic reprogramming of CAR T cells via IDH1 NM OE phenocopies TET2 disruption. Pre-clinical mouse studies are ongoing and will be presented at the ASH.
Disclosures: Melenhorst: Poseida Therapeutics: Membership on an entity's Board of Directors or advisory committees; Janssen Global Services, LLC: Consultancy; IASO Biotherapeutics: Consultancy; Biomarkers: Patents & Royalties.