Exosome-Mediated <i>RUNX3</i> DNA Delivery for Lung Cancer Therapy

Gene therapy represents a promising strategy for treating lung cancer, with the potential to inhibit the proliferation of cancerous cells and induce apoptosis. However, current gene therapy for lung cancer encounters challenges with delivery, targeting, and safety, such as off-target effects, immune responses, and the necessity for better delivery methods. Here, we introduce gene therapy using the key regulator in lung adenocarcinoma, <i>runt-related transcription factor 3</i> (<i>RUNX3</i>), within exosomes (Exos), which are known for their biocompatibility and ability to selectively target cancer cells. We packaged the <i>RUNX3</i> plasmid DNA into human exosomes (hExo-Rs), designed to target and induce apoptosis in cancer cells, resulting in a viability decrease to 43.3%. Normal fibroblasts remained viable at 96.0%, confirming the safety of hExo-Rs for future therapies. We delivered hExo-Rs to cancer spheroids, examined their effects, and found that cytokines from treated cells promote M1 macrophage polarization, emphasizing their potential for immunotherapy. We developed a hydrogel platform for the targeted 14-day release of <i>RUNX3</i> pDNA by attaching hExo-Rs to gelatin using microbial transglutaminase, which enables the selective decrease in cancer cell viability and confirms apoptosis. Our demonstration of <i>RUNX3</i> gene therapy with Exos presents selective anticancer effectiveness and the promise of clinical use through localized, sustained release using the hydrogel.