Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomics

Detalhes bibliográficos
Autor(a) principal: Silvano, Marco
Data de Publicação: 2023
Outros Autores: Virgolini, Nikolaus, Correia, Ricardo, Clarke, Colin, Isidro, Inês A., Alves, Paula M., Roldão, António
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/158521
Resumo: Funding Information: This work has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No. 813453 and the grant agreement N° 730964, and by Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through the following initiatives: “Investigador FCT” Program (IF/01704/2014), Exploratory Research and Development Project (IF/01704/2014/CP1229/CT0001), PhD fellowships (SFRH/BD/134107/2017), iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). Funding from INTERFACE Programme, through the Innovation, Technology, and Circular Economy Fund (FITEC), is gratefully acknowledged. Publisher Copyright: Copyright © 2023 Silvano, Virgolini, Correia, Clarke, Isidro, Alves and Roldão.
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spelling Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomicscell heterogeneityIC-BEVSinfluenza VLPpathway analysissingle-cell RNA sequencingBiotechnologyBioengineeringHistologyBiomedical EngineeringFunding Information: This work has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No. 813453 and the grant agreement N° 730964, and by Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through the following initiatives: “Investigador FCT” Program (IF/01704/2014), Exploratory Research and Development Project (IF/01704/2014/CP1229/CT0001), PhD fellowships (SFRH/BD/134107/2017), iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). Funding from INTERFACE Programme, through the Innovation, Technology, and Circular Economy Fund (FITEC), is gratefully acknowledged. Publisher Copyright: Copyright © 2023 Silvano, Virgolini, Correia, Clarke, Isidro, Alves and Roldão.The insect cell-baculovirus expression vector system (IC-BEVS) has been widely used to produce recombinant protein at high titers, including complex virus-like particles (VPLs). However, cell-to-cell variability upon infection is yet one of the least understood phenomena in virology, and little is known about its impact on production of therapeutic proteins. This study aimed at dissecting insect cell population heterogeneity during production of influenza VLPs in IC-BEVS using single-cell RNA-seq (scRNA-seq). High Five cell population was shown to be heterogeneous even before infection, with cell cycle being one of the factors contributing for this variation. In addition, infected insect cells were clustered according to the timing and level of baculovirus genes expression, with each cluster reporting similar influenza VLPs transgenes (i.e., hemagglutinin and M1) transcript counts. Trajectory analysis enabled to track infection progression throughout pseudotime. Specific pathways such as translation machinery, protein folding, sorting and degradation, endocytosis and energy metabolism were identified as being those which vary the most during insect cell infection and production of Influenza VLPs. Overall, this study lays the ground for the application of scRNA-seq in IC-BEVS processes to isolate relevant biological mechanisms during recombinant protein expression towards its further optimization.Instituto de Tecnologia Química e Biológica António Xavier (ITQB)RUNSilvano, MarcoVirgolini, NikolausCorreia, RicardoClarke, ColinIsidro, Inês A.Alves, Paula M.Roldão, António2023-09-30T22:18:32Z20232023-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/158521eng2296-4185PURE: 72109755https://doi.org/10.3389/fbioe.2023.1143255info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:40:58Zoai:run.unl.pt:10362/158521Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:57:11.090285Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomics
title Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomics
spellingShingle Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomics
Silvano, Marco
cell heterogeneity
IC-BEVS
influenza VLP
pathway analysis
single-cell RNA sequencing
Biotechnology
Bioengineering
Histology
Biomedical Engineering
title_short Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomics
title_full Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomics
title_fullStr Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomics
title_full_unstemmed Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomics
title_sort Dissecting insect cell heterogeneity during influenza VLP production using single-cell transcriptomics
author Silvano, Marco
author_facet Silvano, Marco
Virgolini, Nikolaus
Correia, Ricardo
Clarke, Colin
Isidro, Inês A.
Alves, Paula M.
Roldão, António
author_role author
author2 Virgolini, Nikolaus
Correia, Ricardo
Clarke, Colin
Isidro, Inês A.
Alves, Paula M.
Roldão, António
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
RUN
dc.contributor.author.fl_str_mv Silvano, Marco
Virgolini, Nikolaus
Correia, Ricardo
Clarke, Colin
Isidro, Inês A.
Alves, Paula M.
Roldão, António
dc.subject.por.fl_str_mv cell heterogeneity
IC-BEVS
influenza VLP
pathway analysis
single-cell RNA sequencing
Biotechnology
Bioengineering
Histology
Biomedical Engineering
topic cell heterogeneity
IC-BEVS
influenza VLP
pathway analysis
single-cell RNA sequencing
Biotechnology
Bioengineering
Histology
Biomedical Engineering
description Funding Information: This work has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No. 813453 and the grant agreement N° 730964, and by Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through the following initiatives: “Investigador FCT” Program (IF/01704/2014), Exploratory Research and Development Project (IF/01704/2014/CP1229/CT0001), PhD fellowships (SFRH/BD/134107/2017), iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). Funding from INTERFACE Programme, through the Innovation, Technology, and Circular Economy Fund (FITEC), is gratefully acknowledged. Publisher Copyright: Copyright © 2023 Silvano, Virgolini, Correia, Clarke, Isidro, Alves and Roldão.
publishDate 2023
dc.date.none.fl_str_mv 2023-09-30T22:18:32Z
2023
2023-01-01T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/158521
url http://hdl.handle.net/10362/158521
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 2296-4185
PURE: 72109755
https://doi.org/10.3389/fbioe.2023.1143255
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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