Proteomic Landscape of Adeno-Associated Virus (AAV)-Producing HEK293 Cells
| Autor: | Felipe Guapo, Niall Barron, Jonathan Bones, Nicholas Donohue, Lisa Strasser, Alana Rainbow-Fletcher, Stefano Boi |
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| Rok vydání: | 2021 |
| Předmět: |
Proteome
QH301-705.5 Genetic enhancement Genetic Vectors Gene Expression adeno-associated virus (AAV) Biology Endocytosis Proteomics medicine.disease_cause Transfection Article Catalysis Cell Line Inorganic Chemistry proteomics Tandem Mass Spectrometry medicine Humans Biology (General) LC-MS/MS label-free quantitation Physical and Theoretical Chemistry QD1-999 Molecular Biology Gene Adeno-associated virus Spectroscopy Chromatography Reverse-Phase Organic Chemistry HEK 293 cells SP3 General Medicine Genetic Therapy Dependovirus gene therapy Recombinant Proteins Computer Science Applications Cell biology Chemistry Gene Ontology HEK293 Cells Gene Expression Regulation Proteolysis Lysosomes Signal Transduction |
| Zdroj: | International Journal of Molecular Sciences Volume 22 Issue 21 International Journal of Molecular Sciences, Vol 22, Iss 11499, p 11499 (2021) |
| ISSN: | 1422-0067 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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