Cascade Enzymes Confined\nin DNA Nanoanchors for Antitumor\nTherapy

Cascade-enzyme reaction systems have emerged as promising\ntools\nfor treating malignant tumors by efficiently converting nutrients\ninto toxic substances. However, the challenges of poor localized retention\ncapacity and utilization of highly active enzymes often result in\nextratumoral toxicity and reduced therapeutic efficacy. In this study,\nwe introduced a cell membrane–DNA nanoanchor (DNANA) with a\nspatially confined cascade enzyme for in vivo tumor therapy. The DNANAs\nare constructed using a polyvalent cholesterol-labeled DNA triangular\nprism, ensuring high stability in cell membrane attachment. Glucose\noxidase (GOx) and horseradish peroxidase (HRP), both modified with\nstreptavidin, are precisely confined to biotin-labeled DNANAs. Upon\nintratumoral injection, DNANA enzymes efficiently colonize the tumor\nsite through cellular membrane engineering strategies, significantly\nreducing off-target enzyme leakage and the associated risks of extratumoral\ntoxicity. Furthermore, DNANA enzymes demonstrated effective cancer\ntherapy in vitro and in vivo by depleting glucose and producing highly\ncytotoxic hydroxyl radicals in the vicinity of tumor cells. This membrane-engineered\ncascade-enzyme reaction system presents a conceptual approach to tumor\ntreatment.