Nanoscale Organization of TRAIL Trimers using DNA Origami to Promote Clustering of Death Receptor and Cancer Cell Apoptosis

Abstract Through inducing death receptor (DR) clustering to activate downstream signaling, tumor necrosis factor related apoptosis inducing ligand (TRAIL) trimers trigger apoptosis of tumor cells. However, the poor agonistic activity of current TRAIL‐based therapeutics limits their antitumor efficiency. The nanoscale spatial organization of TRAIL trimers at different interligand distances is still challenging, which is essential for the understanding of interaction pattern between TRAIL and DR. In this study, a flat rectangular DNA origami is employed as display scaffold, and an “engraving‐printing” strategy is developed to rapidly decorate three TRAIL monomers onto its surface to form DNA‐TRAIL3 trimer (DNA origami with surface decoration of three TRAIL monomers). With the spatial addressability of DNA origami, the interligand distances are precisely controlled from 15 to 60 nm. Through comparing the receptor affinity, agonistic activity and cytotoxicity of these DNA‐TRAIL3 trimers, it is found that ≈40 nm is the critical interligand distance of DNA‐TRAIL3 trimers to induce death receptor clustering and the resulting apoptosis.Finally, a hypothetical “active unit” model is proposed for the DR5 clustering induced by DNA‐TRAIL3 trimers.