Single-day nonactivated IL-18–armed CAR T cells establish a durable, stemlike state with enhanced persistence
Abstract
Abstract Chimeric antigen receptor (CAR) T-cell therapies have transformed the treatment of B-cell malignancies, yet challenges including manufacturing delays, T-cell exhaustion, and limited persistence impede broader clinical success. Here, we report the single-day production of nonactivated CAR T cells engineered to secrete interleukin-18 (IL-18), a proinflammatory cytokine that enhances T-cell function. These nonactivated CART19-IL-18 cells (IL-18–secreting anti-CD19 CAR T cells) exhibit robust antitumor efficacy across xenograft models of lymphoma, leukemia, and pancreatic cancer. IL-18 expression enhances the functional advantages of naïve-like nonactivated CAR T cells, resulting in improved persistence, metabolic fitness, and resistance to exhaustion. Single-cell transcriptomic analysis revealed upregulation of IL7R, KLF2, and MCL1, alongside suppression of inhibitory checkpoint genes such as PDCD1, TOX, and HAVCR2. Metabolomic profiling demonstrated enhanced mitochondrial bioenergetics, with increased spare respiratory capacity and accumulation of α-ketoglutarate, malate, and spermine. Functional in vitro and in vivo profiling demonstrated enhanced per-cell cytotoxicity and in vivo durability. We complemented these studies with single-cell transcriptomic and metabolomic analyses to define CAR T-cell biological states beyond what is captured by xenograft tumor clearance. This IL-18–enhanced, activation-free CAR T-cell product offers a clinically actionable platform with the potential to reduce vein-to-vein time while improving product potency and persistence, providing a rationale for clinical testing in patients with tumors refractory to standard CAR T-cell therapy.
Article Details
Authors (18)
Joseph S. Durgin
1Department of Dermatology, University of Michigan, Ann Arbor, MI
Shin Seo
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Shadab Kazmi
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Bakir Valentic
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Chloe Leff
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Martina Markovska
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Xiaoling Jin
Feng Shen
Abdulla Berjis
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Nandana Mukherjee
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Ashwin Sannecy
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Gabriela Plesa
Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia
Khatuna Gabunia
John Scholler
Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia
Saar I. Gill
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Roddy S. O’Connor
3Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA
Carl H. June
Saba Ghassemi
Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia