In Silico Design of Exosome-Mediated CRISPR/Cas۹ Delivery for Dual- Targeting NRF۲/SLC۷A۱۱ in Glioblastoma: A Precision Oncology Approach

Glioblastoma (GBM) remains a formidable challenge in neuro-oncology, characterized by its high aggressiveness and resistance to standard therapies (surgery, radiation, temozolomide) owing to factors like the blood-brain barrier (BBB), tumor heterogeneity, and potent endogenous antioxidant systems [۱]. A promising therapeutic avenue lies in exploiting ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation [۲]. The central axis regulating ferroptosis involves the cystine-glutamate antiporter System Xc (encoded by SLC۷A۱۱), which imports cystine for glutathione (GSH) synthesis, a substrate for glutathione peroxidase ۴ (GPX۴), a key ferroptosis suppressor [۳]. The transcription factor NRF۲ (NFE۲L۲) plays a crucial role in maintaining cellular redox homeostasis by upregulating antioxidant genes, including SLC۷A۱۱ [۴]. Consequently, simultaneous disruption of both NRF۲ and SLC۷A۱۱ presents a compelling strategy to weaken complementary antioxidant defenses by targeting both the upstream transcriptional regulator and the downstream effector of cystine import. Here, we hypothesize that delivering CRISPR/Cas۹ machinery for the simultaneous knockout of NFE۲L۲ and SLC۷A۱۱ via engineered exosomes will synergistically sensitize GBM cells to ferroptosis [۷,۸]. This approach aims to collapse these vital antioxidant layers, thereby inducing cell death and potentially overcoming therapeutic resistance. This study outlines a computational framework to guide the design and optimization of this dual-targeting strategy.