RT Journal Article
SR Electronic
T1 Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.11.28.470258
DO 10.1101/2021.11.28.470258
A1 Emily K. Meseck
A1 Ghiabe Guibinga
A1 Stephen Wang
A1 Cameron McElroy
A1 Eloise Hudry
A1 Keith Mansfield
YR 2021
UL http://biorxiv.org/content/early/2021/11/28/2021.11.28.470258.abstract
AB Biodistribution of self-complementary adeno-associated virus-9 (scAAV9)–chicken β 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/tissues by droplet digital polymerase chain reaction 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 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 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 combination may be better suited for diseases that express a secreted protein and/or do not require widespread brain neuronal transduction.Competing Interest StatementEmily K. Meseck and Cameron McElroy are employees of Novartis Pharmaceuticals Corporation, East Hanover, NJ, USA. Ghiabe Guibinga is an employee of Novartis Institutes for BioMedical Research Biologics Center, San Diego, CA, USA. Stephen Wang, Eloise Hudry, and Keith Mansfield are employees of Novartis Institutes for BioMedical Research, Cambridge, MA, USA.