Engineering Neurotoxin-Functionalized Exosomes for Targeted Delivery to the Peripheral Nervous System

Authors

Mena Asha Krishnan, University of Nebraska Medical CenterFollow
Olawale A. Alimi, University of Nebraska Medical CenterFollow
Tianshu Pan, University of Nebraska Medical CenterFollow
Mitchell Kuss, University of Nebraska Medical CenterFollow
Zeljka Korade, University of Nebraska Medical Center cFollow
Guoku Hu, University of Nebraska Medical CenterFollow
Bo Liu, University of Nebraska Medical CenterFollow
Bin Duan, University of Nebraska Medical CenterFollow

Document Type

Article

Date of this Version

1-12-2024

Citation

Krishnan, M.A.; Alimi, O.A.; Pan, T.; Kuss, M.; Korade, Z.; Hu, G.; Liu, B.; Duan, B. Engineering Neurotoxin-Functionalized Exosomes for Targeted Delivery to the Peripheral Nervous System. Pharmaceutics 2024, 16, 102. https://doi.org/10.3390/ pharmaceutics16010102

Comments

Open access.

Abstract

The administration of therapeutics to peripheral nerve tissue is challenging due to the complexities of peripheral neuroanatomy and the limitations imposed by the blood–nerve barrier (BNB). Therefore, there is a pressing need to enhance delivery effectiveness and implement targeted delivery methods. Recently, erythrocyte-derived exosomes (Exos) have gained widespread attention as biocompatible vehicles for therapeutics in clinical applications. However, engineering targeted Exos for the peripheral nervous system (PNS) is still challenging. This study aims to develop a targeted Exo delivery system specifically designed for presynaptic terminals of peripheral nerve tissue. The clostridium neurotoxin, tetanus toxin-C fragment (TTC), was tethered to the surface of red blood cell (RBC)-derived Exos via a facile and efficient bio-orthogonal click chemistry method without a catalyst. Additionally, Cyanine5 (Cy5), a reactive fluorescent tag, was also conjugated to track Exo movement in both in vitro and in vivo models. Subsequently, Neuro-2a, a mouse neuronal cell line, was treated with dye-labeled Exos with/without TTC in vitro, and the results indicated that TTC-Exos exhibited more efficient accumulation along the soma and axonal circumference, compared to their unmodified counterparts. Further investigation, using a mouse model, revealed that within 72 h of intramuscular administration, engineered TTC-Exos were successfully transported into the neuromuscular junction and sciatic nerve tissues. These results indicated that TTC played a crucial role in the Exo delivery system, improving the affinity to peripheral nerves. These promising results underscore the potential of using targeted Exo carriers to deliver therapeutics for treating peripheral neuropathies.