Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial Cells

Detalhes bibliográficos
Autor(a) principal: Cardoso, M. Helena
Data de Publicação: 2023
Outros Autores: Hall, Michael J., Burgoyne, Thomas, Fale, Pedro, Storm, Tina, Escrevente, Cristina, Antas, Pedro, Seabra, Miguel C., Futter, Clare E.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/157752
Resumo: Purpose: To model the in vivo effects of chloroquine on the retinal pigment epithelium in experimentally tractable cell culture systems and determine the effects of mild chloroquine treatment on lysosome function and turnover. Methods: Effects of low-dose chloroquine treatment on lysosomal function and accessibility to newly endocytosed cargo were investigated in primary and embryonic stem cell-derived RPE cells and ARPE19 cells using fluorescence and electron microscopy of fluorescent and gold-labeled probes. Lysosomal protein expression and accumulation were measured by quantitative PCR and Western blotting. Results: Initial chloroquine-induced lysosome neutralization was followed by partial recovery, lysosomal expansion, and accumulation of undegraded endocytic, phagocytic, and autophagic cargo and inhibition of cathepsin D processing. Accumulation of enlarged lysosomes was accompanied by a gradual loss of accessibility of these structures to the endocytic pathway, implying impaired lysosome reformation. Chloroquine-induced accumulation of pro-cathepsin D, as well as the lysosomal membrane protein, LAMP1, was reproduced by treatment with protease inhibitors and preceded changes in lysosomal gene expression. Conclusions: Low-dose chloroquine treatment inhibits lysosome reformation, causing a gradual depletion of lysosomes able to interact with cargo-carrying vacuoles and degrade their content. The resulting accumulation of newly synthesized pro-cathepsin D and LAMP1 reflects inhibition of normal turnover of lysosomal constituents and possibly lysosomes themselves. A better understanding of the mechanisms underlying lysosome reformation may reveal new targets for the treatment of chloroquine-induced retinopathy.
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spelling Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial CellsOphthalmologySensory SystemsCellular and Molecular NeurosciencePurpose: To model the in vivo effects of chloroquine on the retinal pigment epithelium in experimentally tractable cell culture systems and determine the effects of mild chloroquine treatment on lysosome function and turnover. Methods: Effects of low-dose chloroquine treatment on lysosomal function and accessibility to newly endocytosed cargo were investigated in primary and embryonic stem cell-derived RPE cells and ARPE19 cells using fluorescence and electron microscopy of fluorescent and gold-labeled probes. Lysosomal protein expression and accumulation were measured by quantitative PCR and Western blotting. Results: Initial chloroquine-induced lysosome neutralization was followed by partial recovery, lysosomal expansion, and accumulation of undegraded endocytic, phagocytic, and autophagic cargo and inhibition of cathepsin D processing. Accumulation of enlarged lysosomes was accompanied by a gradual loss of accessibility of these structures to the endocytic pathway, implying impaired lysosome reformation. Chloroquine-induced accumulation of pro-cathepsin D, as well as the lysosomal membrane protein, LAMP1, was reproduced by treatment with protease inhibitors and preceded changes in lysosomal gene expression. Conclusions: Low-dose chloroquine treatment inhibits lysosome reformation, causing a gradual depletion of lysosomes able to interact with cargo-carrying vacuoles and degrade their content. The resulting accumulation of newly synthesized pro-cathepsin D and LAMP1 reflects inhibition of normal turnover of lysosomal constituents and possibly lysosomes themselves. A better understanding of the mechanisms underlying lysosome reformation may reveal new targets for the treatment of chloroquine-induced retinopathy.NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)iNOVA4Health - pólo NMSRUNCardoso, M. HelenaHall, Michael J.Burgoyne, ThomasFale, PedroStorm, TinaEscrevente, CristinaAntas, PedroSeabra, Miguel C.Futter, Clare E.2023-09-13T22:17:33Z2023-08-012023-08-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article10application/pdfhttp://hdl.handle.net/10362/157752eng0146-0404PURE: 70254420https://doi.org/10.1167/iovs.64.11.10info: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:RCAAP2024-05-22T18:14:20Zoai:run.unl.pt:10362/157752Portal AgregadorONGhttps://www.rcaap.pt/oai/openairemluisa.alvim@gmail.comopendoar:71602024-05-22T18:14:20Repositório 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 Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial Cells
title Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial Cells
spellingShingle Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial Cells
Cardoso, M. Helena
Ophthalmology
Sensory Systems
Cellular and Molecular Neuroscience
title_short Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial Cells
title_full Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial Cells
title_fullStr Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial Cells
title_full_unstemmed Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial Cells
title_sort Impaired Lysosome Reformation in Chloroquine-Treated Retinal Pigment Epithelial Cells
author Cardoso, M. Helena
author_facet Cardoso, M. Helena
Hall, Michael J.
Burgoyne, Thomas
Fale, Pedro
Storm, Tina
Escrevente, Cristina
Antas, Pedro
Seabra, Miguel C.
Futter, Clare E.
author_role author
author2 Hall, Michael J.
Burgoyne, Thomas
Fale, Pedro
Storm, Tina
Escrevente, Cristina
Antas, Pedro
Seabra, Miguel C.
Futter, Clare E.
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)
iNOVA4Health - pólo NMS
RUN
dc.contributor.author.fl_str_mv Cardoso, M. Helena
Hall, Michael J.
Burgoyne, Thomas
Fale, Pedro
Storm, Tina
Escrevente, Cristina
Antas, Pedro
Seabra, Miguel C.
Futter, Clare E.
dc.subject.por.fl_str_mv Ophthalmology
Sensory Systems
Cellular and Molecular Neuroscience
topic Ophthalmology
Sensory Systems
Cellular and Molecular Neuroscience
description Purpose: To model the in vivo effects of chloroquine on the retinal pigment epithelium in experimentally tractable cell culture systems and determine the effects of mild chloroquine treatment on lysosome function and turnover. Methods: Effects of low-dose chloroquine treatment on lysosomal function and accessibility to newly endocytosed cargo were investigated in primary and embryonic stem cell-derived RPE cells and ARPE19 cells using fluorescence and electron microscopy of fluorescent and gold-labeled probes. Lysosomal protein expression and accumulation were measured by quantitative PCR and Western blotting. Results: Initial chloroquine-induced lysosome neutralization was followed by partial recovery, lysosomal expansion, and accumulation of undegraded endocytic, phagocytic, and autophagic cargo and inhibition of cathepsin D processing. Accumulation of enlarged lysosomes was accompanied by a gradual loss of accessibility of these structures to the endocytic pathway, implying impaired lysosome reformation. Chloroquine-induced accumulation of pro-cathepsin D, as well as the lysosomal membrane protein, LAMP1, was reproduced by treatment with protease inhibitors and preceded changes in lysosomal gene expression. Conclusions: Low-dose chloroquine treatment inhibits lysosome reformation, causing a gradual depletion of lysosomes able to interact with cargo-carrying vacuoles and degrade their content. The resulting accumulation of newly synthesized pro-cathepsin D and LAMP1 reflects inhibition of normal turnover of lysosomal constituents and possibly lysosomes themselves. A better understanding of the mechanisms underlying lysosome reformation may reveal new targets for the treatment of chloroquine-induced retinopathy.
publishDate 2023
dc.date.none.fl_str_mv 2023-09-13T22:17:33Z
2023-08-01
2023-08-01T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/157752
url http://hdl.handle.net/10362/157752
dc.language.iso.fl_str_mv eng
language eng
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PURE: 70254420
https://doi.org/10.1167/iovs.64.11.10
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eu_rights_str_mv openAccess
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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
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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 mluisa.alvim@gmail.com
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