DNA Origami Nanostructures Elicit Dose‐Dependent Immunogenicity and Are Nontoxic up to High Doses In Vivo

DNA origami (DO) nanotechnology enables the construction of precise nanostructures capable of functionalization with small molecule drugs, nucleic acids, and proteins, suggesting a promising platform for biomedical applications. Despite the potential for drug and vaccine delivery, the impact of DO vehicles on immunogenicity in vivo is not well understood. Here, two DO vehicles, a flat triangle and a nanorod, at varying concentrations are evaluated in vitro and with a repeated dosing regimen administered at a high dose in vivo to study early and late immunogenicity. The studies show normal CD11b+ myeloid cell populations preferentially internalize DO in vitro. DO structures distribute well systemically in vivo, elicit a modest pro‐inflammatory immune response that diminishes over time and are nontoxic as shown by weight, histopathology, lack of cytokine storm, and a complete biochemistry panel at the day 10 end point. The results take critical steps to characterize the biological response to DO and suggest that DO vehicles represent a promising platform for drug delivery and vaccine development where immunogenicity should be a key consideration. Although DNA origami (DO) nanostructures represent an exciting platform for drug delivery, the impact on in vivo immunogenicity at high doses remains unclear. The immune response in vitro and in vivo, and biodistribution profiles are evaluated. DO nanostructures are mildly immunogenic, distribute well, and are nontoxic at the day 10 end point suggesting a promising platform for preclinical drug development.