Meseck, Emily, Mansfield, Keith, Hudry, Eloise, McElroy, Cameron and Guibinga, Ghiabe (2022) Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques. Toxicologic pathology.

Abstract

Biodistribution of self-complementary adeno-associated virus-9 (scAAV9)–chicken β-actin promoter–green fluorescent protein (GFP) was assessed in juvenile cynomolgus macaques infused intrathecally via lumbar puncture or the intracisterna magna (1.0×1013 or 3.0×1013 vg/animal), with necropsy 28 days later. Our results characterized central nervous system biodistribution compared with systemic organs and tissues by droplet digital polymerase chain reaction (ddPCR) for DNA and in situ hybridization. GFP expression was characterized by Meso Scale Discovery electrochemiluminescence immunosorbent assay and immunohistochemistry (IHC). Biodistribution was widespread but variable, with vector DNA and GFP expression greatest in the spinal cord, dorsal root ganglia (DRG), and certain systemic tissues (e.g. liver), with low concentrations detected in many brain regions. Transduction and expression were observed primarily in perivascular astrocytes in the brain, with a paucity of transduction or expression in the neurons. Despite direct administration into the cerebrospinal fluid (CSF), limited distribution to key brain structures identified as targets for monogenic neurologic disease was observed. Greater GFP expression was observed in hepatocytes, striated myocytes (skeletal muscle), cardiomyocytes, spinal cord lower motor neurons, and DRG sensory neurons by IHC. These results suggest caution for use of scAAV9-based intrathecal delivery with the current expression cassette as a modality for neurologic diseases that require widespread brain neuronal expression. This capsid/expression cassette may be better suited for diseases that express a secreted protein and/or do not required widespread brain neuronal transduction.

Item Type:	Article
Date Deposited:	18 Jun 2022 00:45
Last Modified:	18 Jun 2022 00:45
URI:	https://oak.novartis.com/id/eprint/46431