A comparison of AAV-vector production methods for gene therapy and preclinical assessment

Abstract Adeno Associated Virus (AAV)-mediated gene expression in the brain is widely applied in the preclinical setting to investigate the therapeutic potential of specific molecular targets, characterize various cellular functions, and model central nervous system (CNS) diseases. In therapeutic ap...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Marcus Davidsson, Matilde Negrini, Swantje Hauser, Alexander Svanbergsson, Marcus Lockowandt, Giuseppe Tomasello, Fredric P. Manfredsson, Andreas Heuer
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/74a1356fd99448379cbe95576b741883
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:74a1356fd99448379cbe95576b741883
record_format	dspace
spelling	oai:doaj.org-article:74a1356fd99448379cbe95576b7418832021-12-02T11:43:58ZA comparison of AAV-vector production methods for gene therapy and preclinical assessment10.1038/s41598-020-78521-w2045-2322https://doaj.org/article/74a1356fd99448379cbe95576b7418832020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78521-whttps://doaj.org/toc/2045-2322Abstract Adeno Associated Virus (AAV)-mediated gene expression in the brain is widely applied in the preclinical setting to investigate the therapeutic potential of specific molecular targets, characterize various cellular functions, and model central nervous system (CNS) diseases. In therapeutic applications in the clinical setting, gene therapy offers several advantages over traditional pharmacological based therapies, including the ability to directly manipulate disease mechanisms, selectively target disease-afflicted regions, and achieve long-term therapeutic protein expression in the absence of repeated administration of pharmacological agents. Next to the gold-standard iodixanol-based AAV vector production, we recently published a protocol for AAV production based on chloroform-precipitation, which allows for fast in-house production of small quantities of AAV vector without the need for specialized equipment. To validate our recent protocol, we present here a direct side-by-side comparison between vectors produced with either method in a series of in vitro and in vivo assays with a focus on transgene expression, cell loss, and neuroinflammatory responses in the brain. We do not find differences in transduction efficiency nor in any other parameter in our in vivo and in vitro panel of assessment. These results suggest that our novel protocol enables most standardly equipped laboratories to produce small batches of high quality and high titer AAV vectors for their experimental needs.Marcus DavidssonMatilde NegriniSwantje HauserAlexander SvanbergssonMarcus LockowandtGiuseppe TomaselloFredric P. ManfredssonAndreas HeuerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-11 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Marcus Davidsson Matilde Negrini Swantje Hauser Alexander Svanbergsson Marcus Lockowandt Giuseppe Tomasello Fredric P. Manfredsson Andreas Heuer A comparison of AAV-vector production methods for gene therapy and preclinical assessment
description	Abstract Adeno Associated Virus (AAV)-mediated gene expression in the brain is widely applied in the preclinical setting to investigate the therapeutic potential of specific molecular targets, characterize various cellular functions, and model central nervous system (CNS) diseases. In therapeutic applications in the clinical setting, gene therapy offers several advantages over traditional pharmacological based therapies, including the ability to directly manipulate disease mechanisms, selectively target disease-afflicted regions, and achieve long-term therapeutic protein expression in the absence of repeated administration of pharmacological agents. Next to the gold-standard iodixanol-based AAV vector production, we recently published a protocol for AAV production based on chloroform-precipitation, which allows for fast in-house production of small quantities of AAV vector without the need for specialized equipment. To validate our recent protocol, we present here a direct side-by-side comparison between vectors produced with either method in a series of in vitro and in vivo assays with a focus on transgene expression, cell loss, and neuroinflammatory responses in the brain. We do not find differences in transduction efficiency nor in any other parameter in our in vivo and in vitro panel of assessment. These results suggest that our novel protocol enables most standardly equipped laboratories to produce small batches of high quality and high titer AAV vectors for their experimental needs.
format	article
author	Marcus Davidsson Matilde Negrini Swantje Hauser Alexander Svanbergsson Marcus Lockowandt Giuseppe Tomasello Fredric P. Manfredsson Andreas Heuer
author_facet	Marcus Davidsson Matilde Negrini Swantje Hauser Alexander Svanbergsson Marcus Lockowandt Giuseppe Tomasello Fredric P. Manfredsson Andreas Heuer
author_sort	Marcus Davidsson
title	A comparison of AAV-vector production methods for gene therapy and preclinical assessment
title_short	A comparison of AAV-vector production methods for gene therapy and preclinical assessment
title_full	A comparison of AAV-vector production methods for gene therapy and preclinical assessment
title_fullStr	A comparison of AAV-vector production methods for gene therapy and preclinical assessment
title_full_unstemmed	A comparison of AAV-vector production methods for gene therapy and preclinical assessment
title_sort	comparison of aav-vector production methods for gene therapy and preclinical assessment
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/74a1356fd99448379cbe95576b741883
work_keys_str_mv	AT marcusdavidsson acomparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT matildenegrini acomparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT swantjehauser acomparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT alexandersvanbergsson acomparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT marcuslockowandt acomparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT giuseppetomasello acomparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT fredricpmanfredsson acomparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT andreasheuer acomparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT marcusdavidsson comparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT matildenegrini comparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT swantjehauser comparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT alexandersvanbergsson comparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT marcuslockowandt comparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT giuseppetomasello comparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT fredricpmanfredsson comparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment AT andreasheuer comparisonofaavvectorproductionmethodsforgenetherapyandpreclinicalassessment
_version_	1718395349251391488

A comparison of AAV-vector production methods for gene therapy and preclinical assessment

Ejemplares similares