Off-The-Shelf Dual Targeted NK Cells For NHL

Limiting relapse is the biggest challenge following autologous transplantation for non-Hodgkin’s lymphoma. Antigen-directed CAR T cell products and other targeted therapies may be changing the field, yet for high-risk lymphomas, new and safer approaches are needed. We will study the safety and efficacy of antigen-directed, off-the-shelf, induced pluripotent stem cell (iPSC)-derived NK cells engineered for dual-antigen targeting. We have optimized the differentiation of iPSC-derived CD34+ cells to highly potent NK cells, scaled production at the University of Minnesota, and can now produce hundreds of doses in a manufacturing run with a clonal iPSC transduced with a high-affinity, non-cleavable CD16 (called hnCD16). In late 2019, we initiated clinical trials testing this engineered iPSC product (designated FT516). The overarching hypothesis for this project is that dual targeting of NK cells with a CAR and through hnCD16 will protect against relapse after auto-transplant for lymphoma and that future gene edits to alter metabolism will enhance adoptive transfer. This hypothesis fits with the themes of this Program, and Project 2 will inform the Program on the development of off-the-shelf NK cells, NK CAR, and the role of membrane IL-15, as well as pre-clinically on whether manipulating metabolism will enhance NK cell therapy. These investigations are supported by the following Specific Aims. Aim 1 will test dual targeting of NK cells to CD19 and CD20 using two mechanisms of action, ADCC and an NK CAR, to prevent relapse after autologous transplantation for lymphoma. We will test a triple gene-modified iPSC-derived NK cell product (FT596) transduced with hnCD16, an NK cell-optimized CD19 CAR, and membrane bound 15/IL-15Ra fusions. FT596 will be combined with rituximab as maintenance therapy to limit relapse after autologous HCT. This trial is FDA-approved and will first establish an MTD at day +30 after engraftment and then, if single dosing is safe, move to day +7 before engraftment with the goal of 3 doses in the first hundred days to prevent relapse. Aim 2 will test enhancement of NK cell metabolic fitness by ARID5B overexpression and CD38 knockout. Our preliminary data shows both adaptive NK cells and CD38 knockout iPSC-derived NK cells resist oxidative stress- induced cell death. ARID5B overexpression, naturally increased in adaptive NK cells, mediates similar effects. We will generate new iPSC lines to build on FT596 with transgenic expression of ARID5B or a CRISPR knockout of CD38. These clones will be evaluated to probe the mechanisms on how they drive NK cell metabolism and augment function. Aim 3 will test whether manipulating ARID5B and CD38 will drive metabolic fitness and enhance FT596 persistence and function in vivo. These pre-clinical trials will inform future clinical trial modifications—all to promote enhanced anti-lymphoma activity by extending transferred NK cell persistence and potency. These metabolic and potency findings will directly inform Project 1 to improve cell targeting and metabolism, Project 3 for cell persistence, and all 3 Projects to define the advantages of autonomous presentation of IL-15.