Spatial control of cytokine-based therapeutics in pancreatic tumors via DNase I-responsive DNA frameworks

The therapeutic potential of immunomodulatory cytokines in cancer treatment is constrained by the fundamental challenge of decoupling their potent antitumor effects from the toxicities associated with systemic exposure. This limitation has motivated the development of strategies to achieve precise spatial control over cytokine bioactivity in vivo. Here, we introduce a platform that capitalizes on organ-specific biochemical environments-using the pancreas-enriched presence of DNase I as a proof-of-concept-to establish DNase I-activated release therapeutics (DART). Using interferon-α (IFN-α) and interleukin-2 (IL-2) as model cytokines, we demonstrate that these potent immunomodulators are functionally inactivated when tethered to a DNA framework (either DNA origami or a short double-stranded DNA construct) and are selectively reactivated upon cleavage by DNase I. In a syngeneic model of pancreatic ductal adenocarcinoma (PDAC)-a clinically challenging and immunologically resistant tumor type-we show that the DART platform enables localized immune activation while minimizing peripheral exposure. This strategy promotes dendritic cell maturation in tumor-draining lymph nodes, enhances tumor CD8⁺ T cell responses, and achieves potent antitumor efficacy with markedly reduced systemic toxicity. Collectively, our findings establish DART as a versatile platform for the spatially controlled activation of cytokine-based therapeutics, with broad potential for application to other functional protein payloads and distinct organ microenvironments.