Spatially confined IL-12 programs CAR-NK through mTORC1 to coordinate antitumor immune networks
Mubin Tarannum, Khanhlinh Dinh, Mila Stanojevic, Maily Nguyen, Marco Campisi, Andreia Maia, Fuguo Liu, Shaobo Yang, Grace C. Birch, Eden Bobilev, Ian Gillanders, Michal Sheffer, Shikha Gupta, Junning Wang, Suthakar Ganapathy, Daniel E. Michaud, Guillermo Nicolas. Dalton, Seema Chugh, Quang-De Nguyen, Cloud P. Paweletz, Prafulla C. Gokhale, Yang Gao, Jose Cancelas, Jennifer L. Guerriero, Toni K. Choueiri, David A. Barbie, Andrew J. Aguirre, Rebecca L. Porter, Ursula A. Matulonis, John Koreth, Catherine J. Wu, Robert J. Soiffer, Jerome Ritz, Jianzhu Chen, Rizwan RomeeAbstract
Solid tumors evade immunotherapy because of immunosuppressive microenvironments that limit immune cell persistence and function. Although interleukin-12 (IL-12) potently activates antitumor immunity, its clinical use has been constrained by systemic toxicity. Here we engineered a CAR natural killer (NK) cell platform that integrated IL-12 signaling to enhance antitumor immunity. IL-12 signaling synergized with CAR activation to sustain mTORC1 activity through convergent Ras-ERK and PI3K pathways, promoting metabolic fitness, autonomous expansion, and sustained effector function. IL-12 also activated bystander NK cells, T cells, and macrophages to remodel the tumor microenvironment. To improve safety, IL-12 was tethered to a collagen-binding A3-domain, restricting its activity to the extracellular matrix and limiting systemic exposure. In ovarian and pancreatic cancer models, matrix-anchored IL-12 CAR NK cells expanded without exogenous cytokines and achieved durable tumor control. These findings suggest that mTORC1-mediated integration of CAR and cytokine signaling can enhance immune fitness and overcome immunosuppressive tumor microenvironments.