Proteosomal degradation impairs transcytosis of AAV vectors from suprachoroidal space to retina

• Published in: Gene therapy Vol. 28; no. 12; pp. 740 - 747
• Main Authors: Ding, Kun, Shen, Jikui, Hackett, Sean, Khan, Mahmood, Campochiaro, Peter A.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.12.2021; Nature Publishing Group
• Subjects: 14; 14/63; 42; 42/44; 692/699; 692/699/375; Biomedical and Life Sciences; Biomedicine; Care and treatment; Cell Biology; Cellular signal transduction; Choroidal Effusions; Degradation; Dependovirus - genetics; Dependoviruses; Epithelial cells; Epithelium; Expression vectors; Gene Expression; Gene Therapy; Genetic aspects; Genetic vectors; Genetic Vectors - genetics; Health aspects; Human Genetics; Humans; Injection; Medical research; Medicine, Experimental; Methods; Nanotechnology; Photoreceptors; Proteinase inhibitors; Proteolysis; Retina; Retina - metabolism; Retinal degeneration; Retinal pigment epithelium; Rhodopsin kinase; Transcytosis; Transduction; Transduction, Genetic; United States
• ISSN: 0969-7128; 1476-5462; 1476-5462
• DOI: 10.1038/s41434-021-00233-1

Suprachoroidal injection provides a new route of delivery for AAV vectors to retinal pigmented epithelial cells and photoreceptors that can be done in an outpatient setting and is less invasive and potentially safer than subretinal injection, the most common route of delivery for ocular gene therapy. After suprachoroidal injection of AAV8 or AAV9 vectors, there is strong transduction of photoreceptors, but it is unclear how vector traverses the retinal pigmented epithelium. In this study, we found that transduction of photoreceptors was significantly increased after suprachoroidal injection of AAV2tYF-CBA-GFP versus AAV2-CBA-GFP vector. Compared with AAV2, AAV2tYF is more resistant to proteosomal degradation. Treatment with protease inhibitors significantly increased photoreceptor transduction after suprachoroidal injection of AAV5-GRK1-GFP. These data suggest that after suprachoroidal injection, AAV vectors access photoreceptors by transcytosis through retinal pigmented epithelial cells during which they are subject to proteosomal degradation, which if suppressed can enhance transduction of photoreceptors.