Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening
Abstract Human brain cells generated by in vitro cell programming provide exciting prospects for disease modeling, drug discovery and cell therapy. These applications frequently require efficient and clinically compliant tools for genetic modification of the cells. Recombinant adeno-associated virus...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-12404-0 |
| _version_ | 1811250849634058240 |
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| author | Lea Jessica Flitsch Kathleen Börner Christian Stüllein Simon Ziegler Vera Sonntag-Buck Ellen Wiedtke Vesselina Semkova Si Wah Christina Au Yeung Julia Schlee Mohamad Hajo Mona Mathews Beatrice Stefanie Ludwig Susanne Kossatz Horst Kessler Dirk Grimm Oliver Brüstle |
| author_facet | Lea Jessica Flitsch Kathleen Börner Christian Stüllein Simon Ziegler Vera Sonntag-Buck Ellen Wiedtke Vesselina Semkova Si Wah Christina Au Yeung Julia Schlee Mohamad Hajo Mona Mathews Beatrice Stefanie Ludwig Susanne Kossatz Horst Kessler Dirk Grimm Oliver Brüstle |
| author_sort | Lea Jessica Flitsch |
| collection | DOAJ |
| description | Abstract Human brain cells generated by in vitro cell programming provide exciting prospects for disease modeling, drug discovery and cell therapy. These applications frequently require efficient and clinically compliant tools for genetic modification of the cells. Recombinant adeno-associated viruses (AAVs) fulfill these prerequisites for a number of reasons, including the availability of a myriad of AAV capsid variants with distinct cell type specificity (also called tropism). Here, we harnessed a customizable parallel screening approach to assess a panel of natural or synthetic AAV capsid variants for their efficacy in lineage-related human neural cell types. We identified common lead candidates suited for the transduction of directly converted, early-stage induced neural stem cells (iNSCs), induced pluripotent stem cell (iPSC)-derived later-stage, radial glia-like neural progenitors, as well as differentiated astrocytic and mixed neuroglial cultures. We then selected a subset of these candidates for functional validation in iNSCs and iPSC-derived astrocytes, using shRNA-induced downregulation of the citrate transporter SLC25A1 and overexpression of the transcription factor NGN2 for proofs-of-concept. Our study provides a comparative overview of the susceptibility of different human cell programming-derived brain cell types to AAV transduction and a critical discussion of the assets and limitations of this specific AAV capsid screening approach. |
| first_indexed | 2024-04-12T16:10:54Z |
| format | Article |
| id | doaj.art-4875817cf7bc4d6497c93a0dff7c11d4 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-12T16:10:54Z |
| publishDate | 2022-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-4875817cf7bc4d6497c93a0dff7c11d42022-12-22T03:25:54ZengNature PortfolioScientific Reports2045-23222022-05-0112111210.1038/s41598-022-12404-0Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screeningLea Jessica Flitsch0Kathleen Börner1Christian Stüllein2Simon Ziegler3Vera Sonntag-Buck4Ellen Wiedtke5Vesselina Semkova6Si Wah Christina Au Yeung7Julia Schlee8Mohamad Hajo9Mona Mathews10Beatrice Stefanie Ludwig11Susanne Kossatz12Horst Kessler13Dirk Grimm14Oliver Brüstle15Institute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital BonnCenter for Infectious Diseases, Virology, Medical Faculty, Heidelberg UniversityCLADIAC GmbHCLADIAC GmbHCenter for Infectious Diseases, Virology, Medical Faculty, Heidelberg UniversityCenter for Infectious Diseases, Virology, Medical Faculty, Heidelberg UniversityInstitute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital BonnInstitute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital BonnInstitute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital BonnInstitute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital BonnLIFE and BRAIN GmbHDepartment of Nuclear Medicine, School of Medicine, Technical University Munich (TUM), University Hospital Klinikum Rechts der Isar and Central Institute for Translational Cancer Research (Transla TUMDepartment of Nuclear Medicine, School of Medicine, Technical University Munich (TUM), University Hospital Klinikum Rechts der Isar and Central Institute for Translational Cancer Research (Transla TUMInstitute for Advanced Study, Department Chemie, Technical University Munich (TUM)Center for Infectious Diseases, Virology, Medical Faculty, Heidelberg UniversityInstitute of Reconstructive Neurobiology, University of Bonn Medical Faculty and University Hospital BonnAbstract Human brain cells generated by in vitro cell programming provide exciting prospects for disease modeling, drug discovery and cell therapy. These applications frequently require efficient and clinically compliant tools for genetic modification of the cells. Recombinant adeno-associated viruses (AAVs) fulfill these prerequisites for a number of reasons, including the availability of a myriad of AAV capsid variants with distinct cell type specificity (also called tropism). Here, we harnessed a customizable parallel screening approach to assess a panel of natural or synthetic AAV capsid variants for their efficacy in lineage-related human neural cell types. We identified common lead candidates suited for the transduction of directly converted, early-stage induced neural stem cells (iNSCs), induced pluripotent stem cell (iPSC)-derived later-stage, radial glia-like neural progenitors, as well as differentiated astrocytic and mixed neuroglial cultures. We then selected a subset of these candidates for functional validation in iNSCs and iPSC-derived astrocytes, using shRNA-induced downregulation of the citrate transporter SLC25A1 and overexpression of the transcription factor NGN2 for proofs-of-concept. Our study provides a comparative overview of the susceptibility of different human cell programming-derived brain cell types to AAV transduction and a critical discussion of the assets and limitations of this specific AAV capsid screening approach.https://doi.org/10.1038/s41598-022-12404-0 |
| spellingShingle | Lea Jessica Flitsch Kathleen Börner Christian Stüllein Simon Ziegler Vera Sonntag-Buck Ellen Wiedtke Vesselina Semkova Si Wah Christina Au Yeung Julia Schlee Mohamad Hajo Mona Mathews Beatrice Stefanie Ludwig Susanne Kossatz Horst Kessler Dirk Grimm Oliver Brüstle Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening Scientific Reports |
| title | Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening |
| title_full | Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening |
| title_fullStr | Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening |
| title_full_unstemmed | Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening |
| title_short | Identification of adeno-associated virus variants for gene transfer into human neural cell types by parallel capsid screening |
| title_sort | identification of adeno associated virus variants for gene transfer into human neural cell types by parallel capsid screening |
| url | https://doi.org/10.1038/s41598-022-12404-0 |
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