Glioblastoma (GBM) is one of the most aggressive brain tumors, with high recurrence rates and poor prognosis even after surgical resection. Systemically administered chemotherapeutics exhibit limited efficacy and induce toxicity in healthy brain tissues, emphasizing the need for localized and sustained gene therapy strategies. In this study, we constructed an adeno-associated virus (AAV) capsid library using a peptide display-based directed evolution approach to identify novel variants with enhanced brain-targeting specificity. The capsid library was evaluated through in vitro screening against glioma-associated cell lines, resulting in the selection of capsid variants exhibiting improved brain tropism and transduction efficiency. To enhance local delivery and reduce non-specific distribution to healthy tissues, the selected capsid variants were incorporated into a chitosan methacrylate (CHIMA)-based hydrogel. CHIMA is a photocurable hydrogel with excellent biocompatibility and mechanical stability, enabling prolonged release of viral vectors at the target site while preserving their biological activity. Notably, this system suppressed off-target distribution to non-neural tissues, including the liver, and significantly enhanced transduction efficiency in GBM cells. These findings suggest that the integration of engineered AAV vectors with functional hydrogels provides a promising platform for localized and sustained gene therapy with minimized off-target effects. This approach is expected to serve as an effective strategy for treating regionally recurrent tumors such as GBM.
