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Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8
Preclinical studies on gene delivery into mouse lymphocytes are often hampered by insufficient activity of lentiviral (LV) and adeno-associated vectors (AAVs) as well as missing tools for cell type selectivity when considering in vivo gene therapy. Here, we selected designed ankyrin repeat proteins...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8531454/ https://www.ncbi.nlm.nih.gov/pubmed/34729380 http://dx.doi.org/10.1016/j.omtm.2021.09.014 |
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author | Michels, Alexander Frank, Annika M. Günther, Dorothee M. Mataei, Mehryad Börner, Kathleen Grimm, Dirk Hartmann, Jessica Buchholz, Christian J. |
author_facet | Michels, Alexander Frank, Annika M. Günther, Dorothee M. Mataei, Mehryad Börner, Kathleen Grimm, Dirk Hartmann, Jessica Buchholz, Christian J. |
author_sort | Michels, Alexander |
collection | PubMed |
description | Preclinical studies on gene delivery into mouse lymphocytes are often hampered by insufficient activity of lentiviral (LV) and adeno-associated vectors (AAVs) as well as missing tools for cell type selectivity when considering in vivo gene therapy. Here, we selected designed ankyrin repeat proteins (DARPins) binding to murine CD8. The top-performing DARPin was displayed as targeting ligand on both vector systems. When used on engineered measles virus (MV) glycoproteins, the resulting mCD8-LV transduced CD8+ mouse lymphocytes with near-absolute (>99%) selectivity. Despite its lower functional titer, mCD8-LV achieved 4-fold higher gene delivery to CD8+ cells than conventional VSV-LV when added to whole mouse blood. Addition of mCD8-LV encoding a chimeric antigen receptor (CAR) specific for mouse CD19 to splenocytes resulted in elimination of B lymphocytes and lymphoma cells. For display on AAV, the DARPin was inserted into the GH2-GH3 loop of the AAV2 capsid protein VP1, resulting in a DARPin-targeted AAV we termed DART-AAV. Stocks of mCD8-AAV contained similar genome copies as AAV2 but were >20-fold more active in gene delivery in mouse splenocytes, while exhibiting >99% specificity for CD8+ cells. These results suggest that receptor targeting can overcome blocks in transduction of mouse splenocytes. |
format | Online Article Text |
id | pubmed-8531454 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-85314542021-11-01 Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8 Michels, Alexander Frank, Annika M. Günther, Dorothee M. Mataei, Mehryad Börner, Kathleen Grimm, Dirk Hartmann, Jessica Buchholz, Christian J. Mol Ther Methods Clin Dev Original Article Preclinical studies on gene delivery into mouse lymphocytes are often hampered by insufficient activity of lentiviral (LV) and adeno-associated vectors (AAVs) as well as missing tools for cell type selectivity when considering in vivo gene therapy. Here, we selected designed ankyrin repeat proteins (DARPins) binding to murine CD8. The top-performing DARPin was displayed as targeting ligand on both vector systems. When used on engineered measles virus (MV) glycoproteins, the resulting mCD8-LV transduced CD8+ mouse lymphocytes with near-absolute (>99%) selectivity. Despite its lower functional titer, mCD8-LV achieved 4-fold higher gene delivery to CD8+ cells than conventional VSV-LV when added to whole mouse blood. Addition of mCD8-LV encoding a chimeric antigen receptor (CAR) specific for mouse CD19 to splenocytes resulted in elimination of B lymphocytes and lymphoma cells. For display on AAV, the DARPin was inserted into the GH2-GH3 loop of the AAV2 capsid protein VP1, resulting in a DARPin-targeted AAV we termed DART-AAV. Stocks of mCD8-AAV contained similar genome copies as AAV2 but were >20-fold more active in gene delivery in mouse splenocytes, while exhibiting >99% specificity for CD8+ cells. These results suggest that receptor targeting can overcome blocks in transduction of mouse splenocytes. American Society of Gene & Cell Therapy 2021-10-01 /pmc/articles/PMC8531454/ /pubmed/34729380 http://dx.doi.org/10.1016/j.omtm.2021.09.014 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Michels, Alexander Frank, Annika M. Günther, Dorothee M. Mataei, Mehryad Börner, Kathleen Grimm, Dirk Hartmann, Jessica Buchholz, Christian J. Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8 |
title | Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8 |
title_full | Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8 |
title_fullStr | Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8 |
title_full_unstemmed | Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8 |
title_short | Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8 |
title_sort | lentiviral and adeno-associated vectors efficiently transduce mouse t lymphocytes when targeted to murine cd8 |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8531454/ https://www.ncbi.nlm.nih.gov/pubmed/34729380 http://dx.doi.org/10.1016/j.omtm.2021.09.014 |
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