Henan-Macquarie Uni Joint Centre for Biomedical Innovation, Academy for Advanced Interdisciplinary Studies, Henan Key Laboratory of Brain Targeted Bio-nanomedicine, School of Life Sciences, Henan University, Kaifeng, Henan 475004, China.
Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Sydney, NSW 2109, Australia.
Sci Adv. 2022 Apr 22;8(16):eabm8011. doi: 10.1126/sciadv.abm8011. Epub 2022 Apr 20.
We designed a unique nanocapsule for efficient single CRISPR-Cas9 capsuling, noninvasive brain delivery and tumor cell targeting, demonstrating an effective and safe strategy for glioblastoma gene therapy. Our CRISPR-Cas9 nanocapsules can be simply fabricated by encapsulating the single Cas9/sgRNA complex within a glutathione-sensitive polymer shell incorporating a dual-action ligand that facilitates BBB penetration, tumor cell targeting, and Cas9/sgRNA selective release. Our encapsulating nanocapsules evidenced promising glioblastoma tissue targeting that led to high PLK1 gene editing efficiency in a brain tumor (up to 38.1%) with negligible (less than 0.5%) off-target gene editing in high-risk tissues. Treatment with nanocapsules extended median survival time (68 days versus 24 days in nonfunctional sgRNA-treated mice). Our new CRISPR-Cas9 delivery system thus addresses various delivery challenges to demonstrate safe and tumor-specific delivery of gene editing Cas9 ribonucleoprotein for improved glioblastoma treatment that may potentially be therapeutically useful in other brain diseases.
我们设计了一种独特的纳米胶囊，用于高效单 CRISPR-Cas9 包封、非侵入性脑递药和肿瘤细胞靶向，为胶质母细胞瘤基因治疗展示了一种有效且安全的策略。我们的 CRISPR-Cas9 纳米胶囊可通过将单个 Cas9/sgRNA 复合物封装在包含双功能配体的谷胱甘肽敏感聚合物壳内来简单地制备，该配体有利于 BBB 穿透、肿瘤细胞靶向和 Cas9/sgRNA 选择性释放。我们的包封纳米胶囊在脑肿瘤中表现出有前景的胶质母细胞瘤组织靶向，导致 PLK1 基因编辑效率高（高达 38.1%），而在高风险组织中脱靶基因编辑可忽略不计（小于 0.5%）。用纳米胶囊治疗可延长中位生存时间（功能性 sgRNA 治疗的小鼠为 24 天，非功能性 sgRNA 治疗的小鼠为 68 天）。因此，我们的新型 CRISPR-Cas9 递药系统解决了各种递药挑战，展示了用于改善胶质母细胞瘤治疗的安全和肿瘤特异性 Cas9 核糖核蛋白递药，可能对其他脑部疾病具有治疗潜力。
