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Theranostics 2024; 14(12):4773-4786. doi:10.7150/thno.98088 This issue Cite

Research Paper

Brain-targeted intranasal delivery of protein-based gene therapy for treatment of ischemic stroke

Jee-Yeon Ryu¹, Christian Cerecedo-Lopez^1,2, Hongkuan Yang^1,3, Ilhwan Ryu^4,5✉, Rose Du^1✉

1. Department of Neurosurgery, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, United States.
2. Department of Surgery, Valley Baptist Medical Center, University of Texas Rio Grande Valley, Harlingen, TX 78550, United States.
3. Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
4. Department of Chemistry, Kookmin University, Seoul 02707, South Korea.
5. Cooperative Center for Research Facilities, Kookmin University, Seoul 02707, South Korea.

✉ Corresponding authors: ihryuac.kr; rduharvard.edu.

Abstract

Gene therapy using a protein-based CRISPR system in the brain has practical limitations due to current delivery systems, especially in the presence of arterial occlusion. To overcome these obstacles and improve stability, we designed a system for intranasal administration of gene therapy for the treatment of ischemic stroke.

Methods: Nanoparticles containing the protein-based CRISPR/dCas9 system targeting Sirt1 were delivered intranasally to the brain in a mouse model of ischemic stroke. The CRISPR/dCas9 system was encapsulated with calcium phosphate (CaP) nanoparticles to prevent them from being degraded. They were then conjugated with β-hydroxybutyrates (bHb) to target monocarboxylic acid transporter 1 (MCT1) in nasal epithelial cells to facilitate their transfer into the brain.

Results: Human nasal epithelial cells were shown to uptake and transfer nanoparticles to human brain endothelial cells with high efficiency in vitro. The intranasal administration of the dCas9/CaP/PEI-PEG-bHb nanoparticles in mice effectively upregulated the target gene, Sirt1, in the brain, decreased cerebral edema and increased survival after permanent middle cerebral artery occlusion. Additionally, we observed no significant in vivo toxicity associated with intranasal administration of the nanoparticles, highlighting the safety of this approach.

Conclusion: This study demonstrates that the proposed protein-based CRISPR-dCas9 system targeting neuroprotective genes in general, and SIRT1 in particular, can be a potential novel therapy for acute ischemic stroke.

Keywords: Intranasal Delivery, Ischemic Stroke, CRISPR/dCas9, Nanoparticles, Gene Editing

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