Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Michels, Alexander, Frank, Annika M., Günther, Dorothee M., Mataei, Mehryad, Börner, Kathleen, Grimm, Dirk, Hartmann, Jessica, Buchholz, Christian J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2021
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
_version_ 1784586862106509312
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
work_keys_str_mv AT michelsalexander lentiviralandadenoassociatedvectorsefficientlytransducemousetlymphocyteswhentargetedtomurinecd8
AT frankannikam lentiviralandadenoassociatedvectorsefficientlytransducemousetlymphocyteswhentargetedtomurinecd8
AT guntherdorotheem lentiviralandadenoassociatedvectorsefficientlytransducemousetlymphocyteswhentargetedtomurinecd8
AT mataeimehryad lentiviralandadenoassociatedvectorsefficientlytransducemousetlymphocyteswhentargetedtomurinecd8
AT bornerkathleen lentiviralandadenoassociatedvectorsefficientlytransducemousetlymphocyteswhentargetedtomurinecd8
AT grimmdirk lentiviralandadenoassociatedvectorsefficientlytransducemousetlymphocyteswhentargetedtomurinecd8
AT hartmannjessica lentiviralandadenoassociatedvectorsefficientlytransducemousetlymphocyteswhentargetedtomurinecd8
AT buchholzchristianj lentiviralandadenoassociatedvectorsefficientlytransducemousetlymphocyteswhentargetedtomurinecd8