VWA3A-derived ependyma promoter drives increased therapeutic protein secretion into the CSF
Recombinant adeno-associated viral vectors (rAAVs) are a promising strategy to treat neurodegenerative diseases because of their ability to infect non-dividing cells and confer long-term transgene expression. Despite an ever-growing library of capsid variants, widespread delivery of AAVs in the adul...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253123001890 |
| _version_ | 1827890687766429696 |
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| author | Ellie M. Carrell Yong Hong Chen Paul T. Ranum Stephanie L. Coffin Larry N. Singh Luis Tecedor Megan S. Keiser Eloise Hudry Bradley T. Hyman Beverly L. Davidson |
| author_facet | Ellie M. Carrell Yong Hong Chen Paul T. Ranum Stephanie L. Coffin Larry N. Singh Luis Tecedor Megan S. Keiser Eloise Hudry Bradley T. Hyman Beverly L. Davidson |
| author_sort | Ellie M. Carrell |
| collection | DOAJ |
| description | Recombinant adeno-associated viral vectors (rAAVs) are a promising strategy to treat neurodegenerative diseases because of their ability to infect non-dividing cells and confer long-term transgene expression. Despite an ever-growing library of capsid variants, widespread delivery of AAVs in the adult central nervous system remains a challenge. We have previously demonstrated successful distribution of secreted proteins by infection of the ependyma, a layer of post-mitotic epithelial cells lining the ventricles of the brain and central column of the spinal cord, and subsequent protein delivery via the cerebrospinal fluid (CSF). Here we define a functional ependyma promoter to enhance expression from this cell type. Using RNA sequencing on human autopsy samples, we identified disease- and age-independent ependyma gene signatures. Associated promoters were cloned and screened as libraries in mouse and rhesus macaque to reveal cross-species function of a human DNA-derived von Willebrand factor domain containing 3A (VWA3A) promoter. When tested in mice, our VWA3A promoter drove strong, ependyma-localized expression of eGFP and increased secreted ApoE protein levels in the CSF by 2–12× over the ubiquitous iCAG promoter. |
| first_indexed | 2024-03-12T21:14:09Z |
| format | Article |
| id | doaj.art-adb40a4cec5b4a3d9ab912e739f69a5c |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-03-12T21:14:09Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-adb40a4cec5b4a3d9ab912e739f69a5c2023-07-30T04:22:21ZengElsevierMolecular Therapy: Nucleic Acids2162-25312023-09-0133296304VWA3A-derived ependyma promoter drives increased therapeutic protein secretion into the CSFEllie M. Carrell0Yong Hong Chen1Paul T. Ranum2Stephanie L. Coffin3Larry N. Singh4Luis Tecedor5Megan S. Keiser6Eloise Hudry7Bradley T. Hyman8Beverly L. Davidson9Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USARaymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USARaymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USARaymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USACenter for Mitochondrial and Epigenomic Medicine, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USARaymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USARaymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USADepartment of Neurology, Massachusetts General Hospital, Boston, MA 02114, USADepartment of Neurology, Massachusetts General Hospital, Boston, MA 02114, USA; Massachusetts Alzheimer’s Disease Research Center, Charlestown, MA 02129, USA; Harvard Medical School, Boston, MA 02115, USARaymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USA; Department of Pathology & Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Corresponding author: Beverly L. Davidson, Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USA.Recombinant adeno-associated viral vectors (rAAVs) are a promising strategy to treat neurodegenerative diseases because of their ability to infect non-dividing cells and confer long-term transgene expression. Despite an ever-growing library of capsid variants, widespread delivery of AAVs in the adult central nervous system remains a challenge. We have previously demonstrated successful distribution of secreted proteins by infection of the ependyma, a layer of post-mitotic epithelial cells lining the ventricles of the brain and central column of the spinal cord, and subsequent protein delivery via the cerebrospinal fluid (CSF). Here we define a functional ependyma promoter to enhance expression from this cell type. Using RNA sequencing on human autopsy samples, we identified disease- and age-independent ependyma gene signatures. Associated promoters were cloned and screened as libraries in mouse and rhesus macaque to reveal cross-species function of a human DNA-derived von Willebrand factor domain containing 3A (VWA3A) promoter. When tested in mice, our VWA3A promoter drove strong, ependyma-localized expression of eGFP and increased secreted ApoE protein levels in the CSF by 2–12× over the ubiquitous iCAG promoter.http://www.sciencedirect.com/science/article/pii/S2162253123001890MT: Delivery StrategiespromoterependymaAAVneurodegenerationcentral nervous system |
| spellingShingle | Ellie M. Carrell Yong Hong Chen Paul T. Ranum Stephanie L. Coffin Larry N. Singh Luis Tecedor Megan S. Keiser Eloise Hudry Bradley T. Hyman Beverly L. Davidson VWA3A-derived ependyma promoter drives increased therapeutic protein secretion into the CSF Molecular Therapy: Nucleic Acids MT: Delivery Strategies promoter ependyma AAV neurodegeneration central nervous system |
| title | VWA3A-derived ependyma promoter drives increased therapeutic protein secretion into the CSF |
| title_full | VWA3A-derived ependyma promoter drives increased therapeutic protein secretion into the CSF |
| title_fullStr | VWA3A-derived ependyma promoter drives increased therapeutic protein secretion into the CSF |
| title_full_unstemmed | VWA3A-derived ependyma promoter drives increased therapeutic protein secretion into the CSF |
| title_short | VWA3A-derived ependyma promoter drives increased therapeutic protein secretion into the CSF |
| title_sort | vwa3a derived ependyma promoter drives increased therapeutic protein secretion into the csf |
| topic | MT: Delivery Strategies promoter ependyma AAV neurodegeneration central nervous system |
| url | http://www.sciencedirect.com/science/article/pii/S2162253123001890 |
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