The glymphatic system delivery enhances the transduction efficiency of AAV1 to brain endothelial cells in adult mice.

Abstract

BACKGROUND:

Recombinant adeno-associated virus (rAAV) is increasingly applied in neuroscience research or gene therapy. However, there is no simple and efficient tool for specific transfection of rAAV into cerebrovascular tissues. It has been reported that fluorescent tracers or beta-amyloid protein can enter the brain through perivascular spaces, named as "glymphatic system". The purpose of this study was to explore whether rAAV could transduce the cerebral vasculature through the glymphatic pathway.

NEW METHOD:

An AAV1-GFP vector suspension (15 μL) was injected into the intracisternal space of anesthetized mice (n = 2) and 5 μl was injected into the bulbus medullae (n = 2). As controls, 15 μl of artificial cerebrospinal fluid (aCSF) was injected into the cisterna magna. The endothelial specific transduction was verified by Glut1 or PDGFRβ immunofluorescent staining. Immunofluorescence images for all groups were captured with a laser microscope.

RESULTS:

It was observed that infection with rAAV1 vectors encoding green fluorescence protein resulted in a successful cerebrovascular transduction when injected into cisterna magna, compared to aCSF or intra-parenchymal injection at 30 days post-transduction in adult mice. In addition, GFP was co-localized with Glut1 based on immuno-fluorescence. These results indicate that glymphatic system delivery enhances the transduction efficiency of AAV1 to brain endothelial cells.

COMPARISON WITH EXISTING METHODS:

The AAV1 vector can simply and efficiently transduce the cerebral endothelial cells through the glymphatic pathway.

CONCLUSION:

The findings of this study reveal that rAAV1-based vectors have high application potential for endothelial-targeted neurologic disease research or gene-based therapies.

• Authors: Gang Wu, Quan Jiang, Tiantian Cui, Xiuxiu Liu, Dongmei Gong, Yixuan Yin, Chengkun Wang, Tiantian Wang, YingMei Lu, Danyan Zhu, Feng Han
• Journal: Journal of neuroscience methods (J Neurosci Methods) Vol. 328 Pg. 108441 (12 01 2019) ISSN: 1872-678X [Electronic] Netherlands
• PMID: 31574288 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2019 Elsevier B.V. All rights reserved.

Chemical References

• Green Fluorescent Proteins

Topics

• Animals
• Cisterna Magna
• Dependovirus
• Endothelial Cells
• Genetic Vectors
• Glymphatic System
• Green Fluorescent Proteins
• Male
• Mice
• Mice, Inbred C57BL
• Optical Imaging
• Parvovirinae
• Transduction, Genetic