Open AccessIntrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques
Author(s) -
Emily K. Meseck,
Ghiabe Guibinga,
Stephen Wang,
Cameron McElroy,
Eloïse Hudry,
Keith G. Mansfield
Publication year - 2022
Publication title -
toxicologic pathology
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 0.613
H-Index - 108
eISSN - 1533-1601
pISSN - 0192-6233
DOI - 10.1177/01926233221101309
Subject(s) - biodistribution , green fluorescent protein , spinal cord , immunohistochemistry , in situ hybridization , pathology , central nervous system , biology , cerebrospinal fluid , microbiology and biotechnology , systemic administration , medicine , gene expression , neuroscience , biochemistry , in vivo , gene , in vitro
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 (1.0×10 13 or 3.0×10 13 vg/animal), with necropsy 28 days later. Our results characterized central nervous system biodistribution compared with systemic organs/tissues by droplet digital polymerase chain reaction for DNA and in situ hybridization. Green fluorescent protein 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 in many brain regions despite direct cerebrospinal fluid administration. Transduction and expression were observed primarily in perivascular astrocytes in the brain, with a paucity in neurons. Greater GFP expression was observed in hepatocytes, striated myocytes, cardiomyocytes, spinal cord lower motor neurons, and DRG sensory neurons by IHC. These results should be considered when evaluating 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 combination may be better suited for diseases that express a secreted protein and/or do not require widespread brain neuronal transduction.