Enhancing glioblastoma virotherapy with a bioluminescent Zika virus NS5 sensor and integrated gene circuit

About This Grant

ABSTRACT Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults, with a median survival of only 15–18 months even with therapy. The poor prognosis reflects the tumor's rapid growth, cellular heterogeneity, and resistance to current treatments. Oncolytic virotherapy has emerged as a promising adjunct, and Zika virus (ZIKV) has unique potential given its natural tropism for glioblastoma stem-like cells. While attenuated ZIKV strains have shown preclinical efficacy in reducing tumor burden, challenges remain regarding safety, incomplete tumor clearance, and limited control over viral replication. To address these limitations, we propose to develop a novel optogenetic gene circuit, BiViSTA (Bioluminescent Viral Sensor and Transcriptional Activator), that detects ZIKV infection and couples it to therapeutic gene expression. BiViSTA leverages a split- luciferase sensor that reconstitutes upon ZIKV NS5 protein binding, producing bioluminescence that activates a light-sensitive transcription factor to drive pro-apoptotic or immunogenic transgenes. This approach uniquely links viral infection to targeted tumor cell destruction, creating a self-amplifying and tumor-restricted therapeutic system. We will pursue three Specific Aims: Aim 1 will engineer and optimize BiViSTA constructs responsive to ZIKV NS5 and validate their function in permissive human cell lines. Aim 2 will integrate BiViSTA into 2-D and 3-D GBM culture models to test selective induction of pro-apoptotic genes and assess enhanced ZIKV-mediated killing. Aim 3 will evaluate tumor-specific activation and therapeutic efficacy in an orthotopic xenograft mouse model of GBM, including rigorous safety assessments. Our multidisciplinary team combines expertise in synthetic biology, neuro-oncology, and viral pathogenesis, supported by strong preliminary data demonstrating a “plug and play” optogenetic system. The proposed work is highly innovative as no existing technology dynamically couples virus infection to therapeutic gene expression in real time. Successful completion will open the door to a transformative strategy that will improve the safety and efficacy of oncolytic virotherapy for GBM that can integrate nonpathogenic ZIKV pseudovirions. Beyond GBM, BiViSTA represents a broadly adaptable platform for treating virus-associated cancers and for dynamically controlling gene expression in basic and translational research.