Quantitative single-cell transcriptome-based ranking of engineered AAVs in human retinal explants
Gene therapy is a rapidly developing field, and adeno-associated viruses (AAVs) are a leading viral-vector candidate for therapeutic gene delivery. Newly engineered AAVs with improved abilities are now entering the clinic. It has proven challenging, however, to predict the translational potential of...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-06-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050122000626 |
| _version_ | 1818206858032185344 |
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| author | Zhouhuan Xi Bilge E. Öztürk Molly E. Johnson Serhan Turunç William R. Stauffer Leah C. Byrne |
| author_facet | Zhouhuan Xi Bilge E. Öztürk Molly E. Johnson Serhan Turunç William R. Stauffer Leah C. Byrne |
| author_sort | Zhouhuan Xi |
| collection | DOAJ |
| description | Gene therapy is a rapidly developing field, and adeno-associated viruses (AAVs) are a leading viral-vector candidate for therapeutic gene delivery. Newly engineered AAVs with improved abilities are now entering the clinic. It has proven challenging, however, to predict the translational potential of gene therapies developed in animal models due to cross-species differences. Human retinal explants are the only available model of fully developed human retinal tissue and are thus important for the validation of candidate AAV vectors. In this study, we evaluated 18 wild-type and engineered AAV capsids in human retinal explants using a recently developed single-cell RNA sequencing (RNA-seq) AAV engineering pipeline (scAAVengr). Human retinal explants retained the same major cell types as fresh retina, with similar expression of cell-specific markers except for a photoreceptor population with altered expression of photoreceptor-specific genes. The efficiency and tropism of AAVs in human explants were quantified with single-cell resolution. The top-performing serotypes, K91, K912, and 7m8, were further validated in non-human primate and human retinal explants. Together, this study provides detailed information about the transcriptome profiles of retinal explants and quantifies the infectivity of leading AAV serotypes in human retina, accelerating the translation of retinal gene therapies to the clinic. |
| first_indexed | 2024-12-12T04:19:42Z |
| format | Article |
| id | doaj.art-0b6055e7153143f08ae34481985f349e |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-12T04:19:42Z |
| publishDate | 2022-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-0b6055e7153143f08ae34481985f349e2022-12-22T00:38:20ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012022-06-0125476489Quantitative single-cell transcriptome-based ranking of engineered AAVs in human retinal explantsZhouhuan Xi0Bilge E. Öztürk1Molly E. Johnson2Serhan Turunç3William R. Stauffer4Leah C. Byrne5Department of Ophthalmology, University of Pittsburgh, PA, USA; Eye Center of Xiangya Hospital, Hunan Key Laboratory of Ophthalmology, Central South University, Changsha, Hunan, ChinaDepartment of Ophthalmology, University of Pittsburgh, PA, USADepartment of Ophthalmology, University of Pittsburgh, PA, USADepartment of Ophthalmology, University of Pittsburgh, PA, USADepartment of Neurobiology, University of Pittsburgh, PA, USADepartment of Ophthalmology, University of Pittsburgh, PA, USA; Department of Neurobiology, University of Pittsburgh, PA, USA; Department of Bioengineering, University of Pittsburgh, PA, USA; Corresponding author Leah C. Byrne, Department of Ophthalmology, University of Pittsburgh, PA, USA.Gene therapy is a rapidly developing field, and adeno-associated viruses (AAVs) are a leading viral-vector candidate for therapeutic gene delivery. Newly engineered AAVs with improved abilities are now entering the clinic. It has proven challenging, however, to predict the translational potential of gene therapies developed in animal models due to cross-species differences. Human retinal explants are the only available model of fully developed human retinal tissue and are thus important for the validation of candidate AAV vectors. In this study, we evaluated 18 wild-type and engineered AAV capsids in human retinal explants using a recently developed single-cell RNA sequencing (RNA-seq) AAV engineering pipeline (scAAVengr). Human retinal explants retained the same major cell types as fresh retina, with similar expression of cell-specific markers except for a photoreceptor population with altered expression of photoreceptor-specific genes. The efficiency and tropism of AAVs in human explants were quantified with single-cell resolution. The top-performing serotypes, K91, K912, and 7m8, were further validated in non-human primate and human retinal explants. Together, this study provides detailed information about the transcriptome profiles of retinal explants and quantifies the infectivity of leading AAV serotypes in human retina, accelerating the translation of retinal gene therapies to the clinic.http://www.sciencedirect.com/science/article/pii/S2329050122000626AAVretinascRNA-seqgene therapyhuman retinal explant |
| spellingShingle | Zhouhuan Xi Bilge E. Öztürk Molly E. Johnson Serhan Turunç William R. Stauffer Leah C. Byrne Quantitative single-cell transcriptome-based ranking of engineered AAVs in human retinal explants Molecular Therapy: Methods & Clinical Development AAV retina scRNA-seq gene therapy human retinal explant |
| title | Quantitative single-cell transcriptome-based ranking of engineered AAVs in human retinal explants |
| title_full | Quantitative single-cell transcriptome-based ranking of engineered AAVs in human retinal explants |
| title_fullStr | Quantitative single-cell transcriptome-based ranking of engineered AAVs in human retinal explants |
| title_full_unstemmed | Quantitative single-cell transcriptome-based ranking of engineered AAVs in human retinal explants |
| title_short | Quantitative single-cell transcriptome-based ranking of engineered AAVs in human retinal explants |
| title_sort | quantitative single cell transcriptome based ranking of engineered aavs in human retinal explants |
| topic | AAV retina scRNA-seq gene therapy human retinal explant |
| url | http://www.sciencedirect.com/science/article/pii/S2329050122000626 |
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