Modulating immune responses by targeting protein aggregation
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10773/28429 |
Resumo: | Protein aggregation is induced by a wide variety of cellular stresses, including amino acid starvation, virus infection, endoplasmic reticulum stress, lipopolysaccharide, and oxidative stress. It has been suggested that altered proteostasis impacts the inflammatory response, but the underlying mechanism between altered proteostasis and inflammation is still poorly understood. Here, we aim to analyse the impact of protein aggregation in the response of human Dendritic Cells, focusing on plasmacytoid dendritic cells (pDCs). The number of aggregates was manipulated in a pDC cell line, CAL-1, by using both autophagy and proteasome inhibitors. Proteasome inhibition in pDCs induced assembly of large p62-based aggregates, together with an increase on IL-1β secretion and cell death, in an irreversible and cell specific manner. The same effects were not observed upon autophagy inhibition nor on a monocytic cell line. To study the mechanism behind the increase in the inflammatory response upon proteasome inhibition, p62-knockout cells were generated using the CRISPR/Cas9 system. Our data suggests that neither p62 nor the NLRP3 inflammasome are required for induction of cell death upon proteasome inhibition in pDCs. Overall, we conclude that proteasome inhibition induces an inflammatory response specific to pDCs. We propose that this effect must be considered when using proteasome inhibitors as potential drugs for the treatment of pDCs derived immune-mediated disorder and thus, mores studies should be done to clarify the outcome of proteasome inhibition on pDCs. |
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Modulating immune responses by targeting protein aggregationpDCInflammatory responseProtein aggregatesDALISProteasomep62InflammasomeProtein aggregation is induced by a wide variety of cellular stresses, including amino acid starvation, virus infection, endoplasmic reticulum stress, lipopolysaccharide, and oxidative stress. It has been suggested that altered proteostasis impacts the inflammatory response, but the underlying mechanism between altered proteostasis and inflammation is still poorly understood. Here, we aim to analyse the impact of protein aggregation in the response of human Dendritic Cells, focusing on plasmacytoid dendritic cells (pDCs). The number of aggregates was manipulated in a pDC cell line, CAL-1, by using both autophagy and proteasome inhibitors. Proteasome inhibition in pDCs induced assembly of large p62-based aggregates, together with an increase on IL-1β secretion and cell death, in an irreversible and cell specific manner. The same effects were not observed upon autophagy inhibition nor on a monocytic cell line. To study the mechanism behind the increase in the inflammatory response upon proteasome inhibition, p62-knockout cells were generated using the CRISPR/Cas9 system. Our data suggests that neither p62 nor the NLRP3 inflammasome are required for induction of cell death upon proteasome inhibition in pDCs. Overall, we conclude that proteasome inhibition induces an inflammatory response specific to pDCs. We propose that this effect must be considered when using proteasome inhibitors as potential drugs for the treatment of pDCs derived immune-mediated disorder and thus, mores studies should be done to clarify the outcome of proteasome inhibition on pDCs.A agregação de proteínas é um processo que pode ser induzido por uma grande variedade de stresses celulares, incluindo privação de aminoácidos, infeção viral, stress do retículo endoplasmático, lipopolissacarídeos, e stress oxidativo. Alterações na proteostase parecem ter um impacto na resposta inflamatória, mas os mecanismos subjacentes a este impacto são ainda pouco conhecidos. Esta tese tem por objetivo analisar de que forma a agregação de proteínas influencia a resposta das células dendríticas humanas, em particular das células dendríticas plasmocitóides (pDCs). Foram utilizados inibidores de autofagia e do proteossoma para manipular o número de agregados na linha celular de pDCs, CAL-1. A inibição do proteossoma em pDCs induziu a formação de agregados constituídos por p62 de grandes dimensões, e levou à secreção de IL-1β e morte celular, de forma irreversível e específica destas células. Os mesmos efeitos não foram observados após inibição da autofagia nem se verificaram numa outra linha celular, monocítica. Para estudar o mecanismo por detrás do aumento da resposta inflamatória após a inibição do proteossoma, foram criadas células knockout para o p62 usando o sistema CRISPR/Cas9. Os nossos resultados sugerem que nem o p62 nem o inflamassoma de NLRP3 são necessários para a indução da morte celular por inibição do proteossoma em pDCs. De forma geral, concluímos que a inibição do proteossoma induz uma resposta inflamatória específica para pDCs. Propomos que esse efeito deve ser tido em consideração quando se utilizem inibidores de proteossoma como potenciais fármacos para o tratamento de distúrbios mediados por pDCs e, portanto, mais estudos devem ser feitos para esclarecer o efeito da inibição do proteossoma em pDCs.2019-122019-12-01T00:00:00Z2020-12-16T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10773/28429engCarvoeiro, Daniela Claudinoinfo: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:RCAAP2024-02-22T11:55:00Zoai:ria.ua.pt:10773/28429Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:00:58.765518Repositório 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 |
Modulating immune responses by targeting protein aggregation |
title |
Modulating immune responses by targeting protein aggregation |
spellingShingle |
Modulating immune responses by targeting protein aggregation Carvoeiro, Daniela Claudino pDC Inflammatory response Protein aggregates DALIS Proteasome p62 Inflammasome |
title_short |
Modulating immune responses by targeting protein aggregation |
title_full |
Modulating immune responses by targeting protein aggregation |
title_fullStr |
Modulating immune responses by targeting protein aggregation |
title_full_unstemmed |
Modulating immune responses by targeting protein aggregation |
title_sort |
Modulating immune responses by targeting protein aggregation |
author |
Carvoeiro, Daniela Claudino |
author_facet |
Carvoeiro, Daniela Claudino |
author_role |
author |
dc.contributor.author.fl_str_mv |
Carvoeiro, Daniela Claudino |
dc.subject.por.fl_str_mv |
pDC Inflammatory response Protein aggregates DALIS Proteasome p62 Inflammasome |
topic |
pDC Inflammatory response Protein aggregates DALIS Proteasome p62 Inflammasome |
description |
Protein aggregation is induced by a wide variety of cellular stresses, including amino acid starvation, virus infection, endoplasmic reticulum stress, lipopolysaccharide, and oxidative stress. It has been suggested that altered proteostasis impacts the inflammatory response, but the underlying mechanism between altered proteostasis and inflammation is still poorly understood. Here, we aim to analyse the impact of protein aggregation in the response of human Dendritic Cells, focusing on plasmacytoid dendritic cells (pDCs). The number of aggregates was manipulated in a pDC cell line, CAL-1, by using both autophagy and proteasome inhibitors. Proteasome inhibition in pDCs induced assembly of large p62-based aggregates, together with an increase on IL-1β secretion and cell death, in an irreversible and cell specific manner. The same effects were not observed upon autophagy inhibition nor on a monocytic cell line. To study the mechanism behind the increase in the inflammatory response upon proteasome inhibition, p62-knockout cells were generated using the CRISPR/Cas9 system. Our data suggests that neither p62 nor the NLRP3 inflammasome are required for induction of cell death upon proteasome inhibition in pDCs. Overall, we conclude that proteasome inhibition induces an inflammatory response specific to pDCs. We propose that this effect must be considered when using proteasome inhibitors as potential drugs for the treatment of pDCs derived immune-mediated disorder and thus, mores studies should be done to clarify the outcome of proteasome inhibition on pDCs. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-12 2019-12-01T00:00:00Z 2020-12-16T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10773/28429 |
url |
http://hdl.handle.net/10773/28429 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
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 |
repository.mail.fl_str_mv |
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1799137665375797248 |