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High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone

The adult mammalian brain entails a reservoir of neural stem cells (NSCs) generating glial cells and neurons. However, NSCs become increasingly quiescent with age, which hampers their regenerative capacity. New means are therefore required to genetically modify adult NSCs for re-enabling endogenous...

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Autores principales: Kremer, Lukas P.M., Cerrizuela, Santiago, Dehler, Sascha, Stiehl, Thomas, Weinmann, Jonas, Abendroth, Heike, Kleber, Susanne, Laure, Alexander, El Andari, Jihad, Anders, Simon, Marciniak-Czochra, Anna, Grimm, Dirk, Martin-Villalba, Ana
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/PMC8427210/
https://www.ncbi.nlm.nih.gov/pubmed/34553001
http://dx.doi.org/10.1016/j.omtm.2021.07.001
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author Kremer, Lukas P.M.
Cerrizuela, Santiago
Dehler, Sascha
Stiehl, Thomas
Weinmann, Jonas
Abendroth, Heike
Kleber, Susanne
Laure, Alexander
El Andari, Jihad
Anders, Simon
Marciniak-Czochra, Anna
Grimm, Dirk
Martin-Villalba, Ana
author_facet Kremer, Lukas P.M.
Cerrizuela, Santiago
Dehler, Sascha
Stiehl, Thomas
Weinmann, Jonas
Abendroth, Heike
Kleber, Susanne
Laure, Alexander
El Andari, Jihad
Anders, Simon
Marciniak-Czochra, Anna
Grimm, Dirk
Martin-Villalba, Ana
author_sort Kremer, Lukas P.M.
collection PubMed
description The adult mammalian brain entails a reservoir of neural stem cells (NSCs) generating glial cells and neurons. However, NSCs become increasingly quiescent with age, which hampers their regenerative capacity. New means are therefore required to genetically modify adult NSCs for re-enabling endogenous brain repair. Recombinant adeno-associated viruses (AAVs) are ideal gene-therapy vectors due to an excellent safety profile and high transduction efficiency. We thus conducted a high-throughput screening of 177 intraventricularly injected barcoded AAV variants profiled by RNA sequencing. Quantification of barcoded AAV mRNAs identified two synthetic capsids, peptide-modified derivative of wild-type AAV9 (AAV9_A2) and peptide-modified derivative of wild-type AAV1 (AAV1_P5), both of which transduce active and quiescent NSCs. Further optimization of AAV1_P5 by judicious selection of the promoter and dose of injected viral genomes enabled labeling of 30%–60% of the NSC compartment, which was validated by fluorescence-activated cell sorting (FACS) analyses and single-cell RNA sequencing. Importantly, transduced NSCs readily produced neurons. The present study identifies AAV variants with a high regional tropism toward the ventricular-subventricular zone (v-SVZ) with high efficiency in targeting adult NSCs, thereby paving the way for preclinical testing of regenerative gene therapy.
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spelling pubmed-84272102021-09-21 High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone Kremer, Lukas P.M. Cerrizuela, Santiago Dehler, Sascha Stiehl, Thomas Weinmann, Jonas Abendroth, Heike Kleber, Susanne Laure, Alexander El Andari, Jihad Anders, Simon Marciniak-Czochra, Anna Grimm, Dirk Martin-Villalba, Ana Mol Ther Methods Clin Dev Original Article The adult mammalian brain entails a reservoir of neural stem cells (NSCs) generating glial cells and neurons. However, NSCs become increasingly quiescent with age, which hampers their regenerative capacity. New means are therefore required to genetically modify adult NSCs for re-enabling endogenous brain repair. Recombinant adeno-associated viruses (AAVs) are ideal gene-therapy vectors due to an excellent safety profile and high transduction efficiency. We thus conducted a high-throughput screening of 177 intraventricularly injected barcoded AAV variants profiled by RNA sequencing. Quantification of barcoded AAV mRNAs identified two synthetic capsids, peptide-modified derivative of wild-type AAV9 (AAV9_A2) and peptide-modified derivative of wild-type AAV1 (AAV1_P5), both of which transduce active and quiescent NSCs. Further optimization of AAV1_P5 by judicious selection of the promoter and dose of injected viral genomes enabled labeling of 30%–60% of the NSC compartment, which was validated by fluorescence-activated cell sorting (FACS) analyses and single-cell RNA sequencing. Importantly, transduced NSCs readily produced neurons. The present study identifies AAV variants with a high regional tropism toward the ventricular-subventricular zone (v-SVZ) with high efficiency in targeting adult NSCs, thereby paving the way for preclinical testing of regenerative gene therapy. American Society of Gene & Cell Therapy 2021-07-16 /pmc/articles/PMC8427210/ /pubmed/34553001 http://dx.doi.org/10.1016/j.omtm.2021.07.001 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Original Article
Kremer, Lukas P.M.
Cerrizuela, Santiago
Dehler, Sascha
Stiehl, Thomas
Weinmann, Jonas
Abendroth, Heike
Kleber, Susanne
Laure, Alexander
El Andari, Jihad
Anders, Simon
Marciniak-Czochra, Anna
Grimm, Dirk
Martin-Villalba, Ana
High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone
title High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone
title_full High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone
title_fullStr High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone
title_full_unstemmed High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone
title_short High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone
title_sort high throughput screening of novel aav capsids identifies variants for transduction of adult nscs within the subventricular zone
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8427210/
https://www.ncbi.nlm.nih.gov/pubmed/34553001
http://dx.doi.org/10.1016/j.omtm.2021.07.001
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