Abstract
Gene-editing technology shows great potential in glioblastoma (GBM) therapy. Due to the complexity of GBM pathogenesis, a single gene-editing-based therapy is unlikely to be successful; therefore, a multi-gene knockout strategy is preferred for effective GBM inhibition. Here, a non-invasive, biodegradable brain-targeted CRISPR/Cas12a nanocapsule is used that simultaneously targeted dual oncogenes, EGFR and PLK1, for effective GBM therapy. This cargo nanoencapsulation technology enables the CRISPR/Cas12a system to achieve extended blood half-life, efficient blood-brain barrier (BBB) penetration, active tumor targeting, and selective release. In U87MG cells, the combinatorial gene editing system resulted in 61% and 33% knockout of EGFR and PLK1, respectively. Following systemic administration, the CRISPR/Cas12a system demonstrated promising brain tumor accumulation that led to extensive EGFR and PLK1 gene editing in both U87MG and patient-derived GSC xenograft mouse models with negligible off-target gene editing detected through NGS. Additionally, CRISPR/Cas12a nanocapsules that concurrently targeted the EGFR and PLK1 oncogenes showed superior tumor growth suppression and significantly improved the median survival time relative to nanocapsules containing single oncogene knockouts, signifying the potency of the multi-oncogene targeting strategy. The findings indicate that utilization of the CRISPR/Cas12a combinatorial gene editing technique presents a practical option for gene therapy in GBM.
Overview
- The study focuses on the potential of gene-editing technology in glioblastoma (GBM) therapy. A multi-gene knockout strategy is preferred for effective GBM inhibition. A non-invasive, biodegradable brain-targeted CRISPR/Cas12a nanocapsule is used that simultaneously targets dual oncogenes, EGFR and PLK1, for effective GBM therapy. The primary objective of the study is to evaluate the efficacy and safety of the combinatorial gene editing system in U87MG cells and patient-derived GSC xenograft mouse models. The study aims to answer the question of whether the multi-oncogene targeting strategy is more effective than single oncogene knockouts in GBM therapy.
Comparative Analysis & Findings
- The study compares the outcomes observed under different experimental conditions or interventions, specifically the combinatorial gene editing system that targets both EGFR and PLK1 oncogenes versus nanocapsules containing single oncogene knockouts. The results showed that the combinatorial gene editing system resulted in 61% and 33% knockout of EGFR and PLK1, respectively, and demonstrated promising brain tumor accumulation that led to extensive EGFR and PLK1 gene editing in both U87MG and patient-derived GSC xenograft mouse models with negligible off-target gene editing detected through NGS. Additionally, CRISPR/Cas12a nanocapsules that concurrently targeted the EGFR and PLK1 oncogenes showed superior tumor growth suppression and significantly improved the median survival time relative to nanocapsules containing single oncogene knockouts. These findings indicate that the multi-oncogene targeting strategy is more effective than single oncogene knockouts in GBM therapy.
Implications and Future Directions
- The study's findings have significant implications for the field of research and clinical practice in GBM therapy. The multi-gene knockout strategy presented in the study is a practical option for gene therapy in GBM, and the non-invasive, biodegradable brain-targeted CRISPR/Cas12a nanocapsule technology enables efficient blood-brain barrier (BBB) penetration, active tumor targeting, and selective release. Future research directions could include further investigation of the safety and efficacy of the combinatorial gene editing system in larger clinical trials, exploration of other oncogenes that could be targeted in combination with EGFR and PLK1, and development of novel nanocapsule technologies that could improve the delivery and efficacy of gene-editing-based therapies in GBM.