Exosome-associated AAV vector as a robust and convenient neuroscience tool

• Published in: Gene therapy Vol. 23; no. 4; pp. 380 - 392
• Main Authors: Hudry, E, Martin, C, Gandhi, S, György, B, Scheffer, D I, Mu, D, Merkel, S F, Mingozzi, F, Fitzpatrick, Z, Dimant, H, Masek, M, Ragan, T, Tan, S, Brisson, A R, Ramirez, S H, Hyman, B T, Maguire, C A
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-04-2016; Nature Publishing Group
• Subjects: 38/35; 38/5; 42/44; 64/60; Adeno-associated virus; Adenoviruses; Animals; Antibodies; Antibodies, Neutralizing - immunology; Biomedical and Life Sciences; Biomedicine; Blood-brain barrier; Blood-Brain Barrier - metabolism; Brain - metabolism; Brain research; Cell Biology; Cell Line; Central nervous system; Cerebellum; Dependovirus - genetics; Dependoviruses; Exosomes; Expression vectors; Gene Expression; Gene Therapy; Gene transfer; Gene Transfer Techniques; Genetic Therapy - methods; Genetic Vectors - genetics; Health aspects; Human Genetics; Humans; Injection; Methods; Mice; Nanotechnology; Nervous system; Neurons; Neurosciences; original-article-enabling-technologies; Patient outcomes; Pelleting; Physiological aspects; Producer cells; Purkinje cells; Signal transduction; Toxicity; Transduction, Genetic; Transgenes; Ultracentrifugation; United States
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2016.11

Adeno-associated virus (AAV) vectors are showing promise in gene therapy trials and have proven to be extremely efficient biological tools in basic neuroscience research. One major limitation to their widespread use in the neuroscience laboratory is the cost, labor, skill and time-intense purification process of AAV. We have recently shown that AAV can associate with exosomes (exo-AAV) when the vector is isolated from conditioned media of producer cells, and the exo-AAV is more resistant to neutralizing anti-AAV antibodies compared with standard AAV. Here, we demonstrate that simple pelleting of exo-AAV from media via ultracentrifugation results in high-titer vector preparations capable of efficient transduction of central nervous system (CNS) cells after systemic injection in mice. We observed that exo-AAV is more efficient at gene delivery to the brain at low vector doses relative to conventional AAV, even when derived from a serotype that does not normally efficiently cross the blood–brain barrier. Similar cell types were transduced by exo-AAV and conventionally purified vector. Importantly, no cellular toxicity was noted in exo-AAV-transduced cells. We demonstrated the utility and robustness of exo-AAV-mediated gene delivery by detecting direct GFP fluorescence after systemic injection, allowing three-dimensional reconstruction of transduced Purkinje cells in the cerebellum using
ex vivo
serial two-photon tomography. The ease of isolation combined with the high efficiency of transgene expression in the CNS, may enable the widespread use of exo-AAV as a neuroscience research tool. Furthermore, the ability of exo-AAV to evade neutralizing antibodies while still transducing CNS after peripheral delivery is clinically relevant.