Reverse Engineering of DNA and RNA Hybrid Origami Structures as Targeted Nanomedicine for KRAS-Mutated Lung Cancer Therapy

Nonsmall cell lung cancer (NSCLC), driven by KRAS gene mutations, is a highly malignant disease currently lacking targeted medicines, except for a specific G12C mutation. However, nanotechnology-based interventions and nanomedicines hold great promise as alternatives to traditional chemotherapy. In this study, we propose a reverse engineering strategy to design and assemble DNA and RNA hybrid origami nanostructures as nanomedicines for the therapy of KRAS-mutated NSCLC. Instead of using M13 DNA as a scaffold for origami design, whose scaffold sequence is constant, the proposed reverse engineering strategy uses a pool of staple sequences that are constant while the scaffold sequences are variable. We conducted a research study on concept verification by designing DNA and RNA hybrid origami nanotubular structures whose staple strands are antisense oligonucleotides (ASON) that are complementary to the full exon regions of KRAS mRNA. The scaffold RNA sequence is thus determined by the ASON sequences and the geometry of the origami design. Once inside the cancer cells, the structure degrades the RNA and releases ASON under the activation of RNase H to exert an antitumor effect. The results from cellular experiments and in vivo studies demonstrated that the hybrid origami structure, composed of DNA and RNA, effectively inhibited both cell proliferation and tumor progression. Remarkably, the proposed reverse engineering method is a universal strategy that can be extended to designing nanomedicines targeting other pathogenic genes and diseases and has good prospects.