Cargando…

Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques

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 da...

Descripción completa

Detalles Bibliográficos
Autores principales: Meseck, Emily K., Guibinga, Ghiabe, Wang, Stephen, McElroy, Cameron, Hudry, Eloise, Mansfield, Keith
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9284083/
https://www.ncbi.nlm.nih.gov/pubmed/35658751
http://dx.doi.org/10.1177/01926233221101309
_version_ 1784747478631841792
author Meseck, Emily K.
Guibinga, Ghiabe
Wang, Stephen
McElroy, Cameron
Hudry, Eloise
Mansfield, Keith
author_facet Meseck, Emily K.
Guibinga, Ghiabe
Wang, Stephen
McElroy, Cameron
Hudry, Eloise
Mansfield, Keith
author_sort Meseck, Emily K.
collection PubMed
description 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.
format Online
Article
Text
id pubmed-9284083
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher SAGE Publications
record_format MEDLINE/PubMed
spelling pubmed-92840832022-07-16 Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques Meseck, Emily K. Guibinga, Ghiabe Wang, Stephen McElroy, Cameron Hudry, Eloise Mansfield, Keith Toxicol Pathol Original Articles 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. SAGE Publications 2022-06-03 2022-06 /pmc/articles/PMC9284083/ /pubmed/35658751 http://dx.doi.org/10.1177/01926233221101309 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by-nc/4.0/This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Original Articles
Meseck, Emily K.
Guibinga, Ghiabe
Wang, Stephen
McElroy, Cameron
Hudry, Eloise
Mansfield, Keith
Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques
title Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques
title_full Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques
title_fullStr Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques
title_full_unstemmed Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques
title_short Intrathecal sc-AAV9-CB-GFP: Systemic Distribution Predominates Following Single-Dose Administration in Cynomolgus Macaques
title_sort intrathecal sc-aav9-cb-gfp: systemic distribution predominates following single-dose administration in cynomolgus macaques
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9284083/
https://www.ncbi.nlm.nih.gov/pubmed/35658751
http://dx.doi.org/10.1177/01926233221101309
work_keys_str_mv AT meseckemilyk intrathecalscaav9cbgfpsystemicdistributionpredominatesfollowingsingledoseadministrationincynomolgusmacaques
AT guibingaghiabe intrathecalscaav9cbgfpsystemicdistributionpredominatesfollowingsingledoseadministrationincynomolgusmacaques
AT wangstephen intrathecalscaav9cbgfpsystemicdistributionpredominatesfollowingsingledoseadministrationincynomolgusmacaques
AT mcelroycameron intrathecalscaav9cbgfpsystemicdistributionpredominatesfollowingsingledoseadministrationincynomolgusmacaques
AT hudryeloise intrathecalscaav9cbgfpsystemicdistributionpredominatesfollowingsingledoseadministrationincynomolgusmacaques
AT mansfieldkeith intrathecalscaav9cbgfpsystemicdistributionpredominatesfollowingsingledoseadministrationincynomolgusmacaques