Spatially Controlled Activation of Toll-like Receptor 9 with DNA-Based Nanomaterials

First evidence of geometrical patterns and defined distances of biomolecules as fundamental parameters to regulate receptor binding and cell signaling have emerged recently. Here, we demonstrate the importance of controlled nanospacing of immunostimulatory agents for the activation of immune cells by exploiting DNA-based nanomaterials and pre-existing crystallography data. We created DNA origami nanoparticles that present CpG-motifs in rationally designed spatial patterns to activate Toll-like Receptor 9 in RAW 264.7 macrophages. We demonstrated that stronger immune activation is achieved when active molecules are positioned at the distance of 7 nm, matching the active dimer structure of the receptor. Moreover, we show how the introduction of linkers between particle and ligand can influence the spatial tolerance of binding. These findings are fundamental for a fine-tuned manipulation of the immune system, considering the importance of spatially controlled presentation of therapeutics to increase efficacy and specificity of immune-modulating nanomaterials where multivalent binding is involved.

Automated summary

The research provides the first evidence of the importance of geometric patterns and defined distances of biomolecules in regulating receptor binding and cell signaling. By utilizing DNA-based nanomaterials and crystallography data, the study demonstrates the significance of controlled nanospacing of immunostimulatory agents for immune cell activation. The study creates DNA origami nanoparticles with spatially patterned CpG motifs to activate Toll-like Receptor 9 in macrophages, showing that stronger immune activation is achieved when active molecules are positioned at a specific distance matching the receptor's active dimer structure. Moreover, the study reveals how the introduction of linkers between nanoparticles and ligands can impact binding.