Proteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells

Adeno-associated viral (AAV) vectors are widely used for gene therapy, providing treatment for diseases caused by absent or defective genes. Despite the success of gene therapy, AAV manufacturing is still challenging, with production yields being limited. With increased patient demand, improvements...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Lisa Strasser, Stefano Boi, Felipe Guapo, Nicholas Donohue, Niall Barron, Alana Rainbow-Fletcher, Jonathan Bones
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	proteomics LC-MS/MS SP3 label-free quantitation HEK293 cells adeno-associated virus (AAV) Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/93e3678698304bf1bc3c91fc10ee7d88
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:93e3678698304bf1bc3c91fc10ee7d88
record_format	dspace
spelling	oai:doaj.org-article:93e3678698304bf1bc3c91fc10ee7d882021-11-11T16:57:44ZProteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells10.3390/ijms2221114991422-00671661-6596https://doaj.org/article/93e3678698304bf1bc3c91fc10ee7d882021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11499https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Adeno-associated viral (AAV) vectors are widely used for gene therapy, providing treatment for diseases caused by absent or defective genes. Despite the success of gene therapy, AAV manufacturing is still challenging, with production yields being limited. With increased patient demand, improvements in host cell productivity through various engineering strategies will be necessary. Here, we study the host cell proteome of AAV5-producing HEK293 cells using reversed phase nano-liquid chromatography and tandem mass spectrometry (RPLC-MS/MS). Relative label-free quantitation (LFQ) was performed, allowing a comparison of transfected vs. untransfected cells. Gene ontology enrichment and pathway analysis revealed differential expression of proteins involved in fundamental cellular processes such as metabolism, proliferation, and cell death. Furthermore, changes in expression of proteins involved in endocytosis and lysosomal degradation were observed. Our data provides highly valuable insights into cellular mechanisms involved during recombinant AAV production by HEK293 cells, thus potentially enabling further improvements of gene therapy product manufacturing.Lisa StrasserStefano BoiFelipe GuapoNicholas DonohueNiall BarronAlana Rainbow-FletcherJonathan BonesMDPI AGarticleproteomicsLC-MS/MSSP3label-free quantitationHEK293 cellsadeno-associated virus (AAV)Biology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11499, p 11499 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	proteomics LC-MS/MS SP3 label-free quantitation HEK293 cells adeno-associated virus (AAV) Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	proteomics LC-MS/MS SP3 label-free quantitation HEK293 cells adeno-associated virus (AAV) Biology (General) QH301-705.5 Chemistry QD1-999 Lisa Strasser Stefano Boi Felipe Guapo Nicholas Donohue Niall Barron Alana Rainbow-Fletcher Jonathan Bones Proteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells
description	Adeno-associated viral (AAV) vectors are widely used for gene therapy, providing treatment for diseases caused by absent or defective genes. Despite the success of gene therapy, AAV manufacturing is still challenging, with production yields being limited. With increased patient demand, improvements in host cell productivity through various engineering strategies will be necessary. Here, we study the host cell proteome of AAV5-producing HEK293 cells using reversed phase nano-liquid chromatography and tandem mass spectrometry (RPLC-MS/MS). Relative label-free quantitation (LFQ) was performed, allowing a comparison of transfected vs. untransfected cells. Gene ontology enrichment and pathway analysis revealed differential expression of proteins involved in fundamental cellular processes such as metabolism, proliferation, and cell death. Furthermore, changes in expression of proteins involved in endocytosis and lysosomal degradation were observed. Our data provides highly valuable insights into cellular mechanisms involved during recombinant AAV production by HEK293 cells, thus potentially enabling further improvements of gene therapy product manufacturing.
format	article
author	Lisa Strasser Stefano Boi Felipe Guapo Nicholas Donohue Niall Barron Alana Rainbow-Fletcher Jonathan Bones
author_facet	Lisa Strasser Stefano Boi Felipe Guapo Nicholas Donohue Niall Barron Alana Rainbow-Fletcher Jonathan Bones
author_sort	Lisa Strasser
title	Proteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells
title_short	Proteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells
title_full	Proteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells
title_fullStr	Proteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells
title_full_unstemmed	Proteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells
title_sort	proteomic landscape of adeno-associated virus (aav)-producing hek293 cells
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/93e3678698304bf1bc3c91fc10ee7d88
work_keys_str_mv	AT lisastrasser proteomiclandscapeofadenoassociatedvirusaavproducinghek293cells AT stefanoboi proteomiclandscapeofadenoassociatedvirusaavproducinghek293cells AT felipeguapo proteomiclandscapeofadenoassociatedvirusaavproducinghek293cells AT nicholasdonohue proteomiclandscapeofadenoassociatedvirusaavproducinghek293cells AT niallbarron proteomiclandscapeofadenoassociatedvirusaavproducinghek293cells AT alanarainbowfletcher proteomiclandscapeofadenoassociatedvirusaavproducinghek293cells AT jonathanbones proteomiclandscapeofadenoassociatedvirusaavproducinghek293cells
_version_	1718432190017044480

Proteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells

Ejemplares similares