Bio-clickable, small extracellular vesicles-COCKTAIL therapy for ischemic stroke

Khan Haroon; Huitong Ruan; Haoran Zheng; Shengju Wu; Ze Liu; Xiaojing Shi; Yaohui Tang; Guo-Yuan Yang; Zhijun Zhang

doi:10.1016/j.jconrel.2023.10.003

Bio-clickable, small extracellular vesicles-COCKTAIL therapy for ischemic stroke

J Control Release. 2023 Nov:363:585-596. doi: 10.1016/j.jconrel.2023.10.003. Epub 2023 Oct 11.

Authors

Khan Haroon¹, Huitong Ruan², Haoran Zheng¹, Shengju Wu¹, Ze Liu¹, Xiaojing Shi³, Yaohui Tang¹, Guo-Yuan Yang⁴, Zhijun Zhang⁵

Affiliations

¹ Shanghai Jiao Tong University Affiliated Sixth People's Hospital, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China.
² Department of Orthopaedics, Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint Diseases, Shanghai Institute of Traumatology and Orthopaedics, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
³ Shanghai Jiao Tong University Affiliated Sixth People's Hospital, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
⁴ Shanghai Jiao Tong University Affiliated Sixth People's Hospital, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China. Electronic address: gyyang@sjtu.edu.cn.
⁵ Shanghai Jiao Tong University Affiliated Sixth People's Hospital, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China. Electronic address: zhangzj@sjtu.edu.cn.

PMID: 37793483
DOI: 10.1016/j.jconrel.2023.10.003

Abstract

Delivering large therapeutic molecules via the blood-brain barrier to treat ischemic stroke remains challenging. NR2B9c is a potent neuroprotective peptide but it's safe and targeted delivery to the brain requires an efficient, natural, and non-immunogenic delivery technique. Small extracellular vesicles (sEVs) have shown great potential as a non-immunogenic, natural cargo delivery system; however, tailoring of its inefficient brain targeting is desired. Here, we coupled rabies virus glycoprotein 29 with sEVs surface via bio-orthogonal click chemistry reactions, followed by loading of NR2B9c, ultimately generating stroke-specific therapeutic COCKTAIL (sEVs-COCKTAIL). Primary neurons and Neuro-2a cells were cultured for in vitro and transient middle cerebral artery occlusion model was used for in vivo studies to evaluate neuron targeting and anti-ischemic stroke potential of the sEVs-COCKTAIL. Bio-clickable sEVs were selectively taken up by neurons but not glial cells. In the in vitro ischemic stroke model of oxygen-glucose deprivation, the sEVs-COCKTAIL exhibited remarkable potential against reactive oxygen species and cellular apoptosis. In vivo studies further demonstrated the brain targeting and increased half-life of bio-clickable sEVs, delivering NR2B9c to the ischemic brain and reducing stroke injury. Treatment with the sEVs-COCKTAIL significantly increased behavioral recovery and reduced neuronal apoptosis after transient middle cerebral artery occlusion. NR2B9c was delivered to neurons binding to post-synaptic density protein-95, inhibiting N-methyl-d-Aspartate receptor-mediated over production of oxidative stress and mitigating protein B-cell lymphoma 2 and P38 proteins expression. Our results provide an efficient and biocompatible approach to a targeted delivery system, which is a promising modality for stroke therapy.

Keywords: Drug delivery; NR2B9c; Rabies virus glycoprotein 29; Small extracellular vesicles; Stroke.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Brain Ischemia* / drug therapy
Brain Ischemia* / metabolism
Extracellular Vesicles* / metabolism
Humans
Infarction, Middle Cerebral Artery / drug therapy
Ischemic Stroke* / drug therapy
Stroke* / drug therapy