Tetrahedral DNA Nanostructures as a Multifactorial Nanotherapeutic Platform for Deep Vein Thrombosis: Targeting Necroptosis, Oxidative Stress, and Inflammation.

Purpose: Venous thromboembolism (VTE), comprising deep vein thrombosis (DVT) and pulmonary embolism (PE), remains a major clinical challenge due to the limitations of conventional anticoagulant therapies. This study explores tetrahedral DNA nanostructures (TDNs) as a novel therapeutic strategy for DVT by targeting endothelial cell necroptosis and oxidative stress. Methods: A mouse DVT model was established via inferior vena cava ligation. TDNs were synthesized from four complementary ssDNA strands and validated via gel electrophoresis and atomic force microscopy. Therapeutic effects were assessed through histopathology, Western blot, qPCR, RNA-seq, and oxidative stress markers (SOD/MDA). Mechanistic insights were explored via transcriptomics, co-immunoprecipitation, and bioinformatics. Results: In a mouse DVT model, TDNs demonstrated remarkable efficacy in mitigating thrombosis, reducing endothelial damage, and restoring vascular homeostasis. Mechanistically, TDNs downregulated the phosphorylation of RIP3 and MLKL, suppressed necroptosis, and modulated inflammatory signaling, specifically via the interaction between PTPRC and CCR9. Transcriptomic analysis confirmed that TDNs ameliorated the dysregulated expression of immune-inflammatory mediators while promoting antioxidative effects by increasing SOD activity and decreasing MDA levels. Bioinformatics and protein interaction assays further unveiled a direct binding between RIP3 and PTPRC, highlighting their roles as molecular targets of TDNs. Conclusion: These findings underscore TDNs' potential as a safe and effective nanotherapeutic platform for managing DVT by simultaneously targeting necroptosis, oxidative stress, and inflammation. Future studies are warranted to optimize dosing strategies and evaluate their long-term safety and synergistic use with established anticoagulants.