Work

120. Tropism and Toxicity of Adeno-Associated Viral Vector Serotypes on Neurons and Glia In Vitro

Year: 2006

Type: article

Abstract: Viral vector toxicity is a primary concern in choosing an optimal gene delivery vehicle for either studying biological phenomena or evaluating potential therapeutic strategies. Generally, adenoassociated viral vectors are thought to be among the least toxic viral vectors. Currently, over 100 serotypes of AAV have been identified or produced and the tropism of the serotypes varies. In the central nervous system, studies have shown serotypes 1, 2, 5, 6, 7, 8, 9 and 10 are capable of transduction. We have previously reported the transduction of primary neuronal cultures by serotypes 1, 2, 5, and 6. Here, we compare the toxicity of serotypes 1, 2, 5, 6, 7, 8 and 9 and tropism of serotypes 7, 8 and 9 on primary cortical cultures derived from rat embryonic brain tissue. A self-complementing AAV vector containing modified AAV2 terminal repeats flanking a green fluorescent protein expressed from the CMV promoter was packaged using the cap gene from AAV serotypes 1, 2, 5, 6, 7, 8, or 9. All vectors were purified by CsCl centrifugation. Cultures were infected on DIV6 then examined for GFP fluorescence, toxicity and immunoreactivity on DIV8 and 12. At 48 hours after infection at a comparable MOI of 10000, only serotype 1 showed significant toxicity as determined by LDH assay. Using an MTS assay, similar toxicity was observed at an MOI of 100000; however, at a higher MOI, serotypes 1, 6, 7 and 8 exhibited greater toxicity than serotypes 2, 5 and 9. One week after infection, serotypes 7 and 8 resulted in primarily NeuN+ colocalized with GFP fluorescence, however, GFAP+ fibers expressing GFP were detected. Serotype 9 infected both NeuN+ and GFAP+ cells. Overall, AAV serotypes 1, 2, 5, 6, 7, 8 and 9 transduce rat neurons and glia in vitro with varied efficiency and toxicity.

Source: Molecular Therapy

DOI: 10.1016/j.ymthe.2006.08.140

Authors: Doug B. Howard, Kathleen Powers, Barry J. Hoffer, Yun Wang, Brandon K. Harvey

Institution: National Institute on Drug Abuse

Language: English

Topic: Virus-based gene therapy research

Subfield: Genetics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: hybrid

Estimated APC paid: $4,500