The AAV9 receptor and its modification to improve in vivo lung gene transfer in mice
Christie L. Bell, … , Mavis Agbandje-McKenna, James M. Wilson
Christie L. Bell, … , Mavis Agbandje-McKenna, James M. Wilson
Published May 16, 2011
Citation Information: J Clin Invest. 2011;121(6):2427-2435. https://doi.org/10.1172/JCI57367.
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Research Article Genetics

The AAV9 receptor and its modification to improve in vivo lung gene transfer in mice

Abstract

Vectors based on adeno-associated virus (AAV) serotype 9 are candidates for in vivo gene delivery to many organs, but the receptor(s) mediating these tropisms have yet to be defined. We evaluated AAV9 uptake by glycans with terminal sialic acids (SAs), a common mode of cellular entry for viruses. We found, however, that AAV9 binding increased when terminal SA was enzymatically removed, suggesting that galactose, which is the most commonly observed penultimate monosaccharide to SA, may mediate AAV9 transduction. This was confirmed in mutant CHO Pro-5 cells deficient in the enzymes involved in glycoprotein biogenesis, as well as lectin interference studies. Binding of AAV9 to glycans with terminal galactose was demonstrated via glycan binding assays. Co-instillation of AAV9 vector with neuraminidase into mouse lung resulted in exposure of terminal galactose on the apical surface of conducting airway epithelial cells, as shown by lectin binding and increased transduction of these cells, demonstrating the possible utility of this vector in lung-directed gene transfer. Increasing the abundance of the receptor on target cells and improving vector efficacy may improve delivery of AAV vectors to their therapeutic targets.

Authors

Christie L. Bell, Luk H. Vandenberghe, Peter Bell, Maria P. Limberis, Guang-Ping Gao, Kim Van Vliet, Mavis Agbandje-McKenna, James M. Wilson

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Figure 5

Expression of galactose in the cells of the murine conducting airways.

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Expression of galactose in the cells of the murine conducting airways. C...
C57BL/6 mice were treated with PBS (A–D) or 100 mU NA (E–H) in a total of 30 μl delivered intranasally. Lungs were inflated and removed 1 hour later, and thin sections (8 μm) were stained with (A, C, E, and G) rhodamine-RCA, (B, D, F, and H) fluorescein-SNA, and (C, D, G, and H) DAPI. Sections were examined by wide-field ×200 magnification (A, B, E, and F) and confocal microscopy (C, D, G, and H). Scale bar: 20 μm. (I–K) Lung sections of mice treated intranasally with NA were stained for (I) galactose expression using rhodamine–RCA lectin and (J) α-tubulin expression to stain cilia. (K) Overlay of galactose and α-tubulin staining. Sections were examined at ×400 magnification.