Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury

Autores
Tadic, Mariana S.; Renna, Felipe Javier; Herrera Lopez, Malena; Orquera, Tamara; Lopez Mingorance, Fabiana Norma; Ropolo, Alejandro Javier; Vaccaro, Maria Ines
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Cellular stress activates various mechanisms, including autophagy and vesicular trafficking, to maintain homeostasis and cope with pathological conditions such as acute pancreatitis. One of these mechanisms involves the unconventional secretion of extracellular vesicles (EVs), lipid bilayer-delimited particles released by almost all cell types into the extracellular medium. Recently, an autophagy-related secretory pathway that releases EVs has been discovered. Our study focuses on Vacuole Membrane Protein 1 (VMP1), an autophagy-related protein implicated in pancreatitis, diabetes, pancreatic cancer, and cellular stress management. VMP1 expression triggers autophagy in mammalian cells through its ubiquitination and its direct binding to BECN1. Here, we demonstrate that VMP1 is secreted to the extracellular medium integrated into the membrane of EVs. Using cell lines expressing different VMP1-tag expression plasmids, we successfully isolated and purified VMP1-containing EVs (VMP1-EVs) from the extracellular medium through ultracentrifugation and immune isolation techniques. The secretion of VMP1-EVs was reduced by mTOR inhibition and in the absence of proteins essential for autophagosome formation, such as ATG5. The secretion of endogenous VMP1 by pancreatic acinar cells was highly induced in experimental models of cell stress, including blocked autophagic flux with Bafilomycin and exposure to supramaximal doses of caerulein. Moreover, immuno-isolation of VMP1-EVs from cell models revealed that VMP1 is a component of EV membranes. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis of immuno-isolated VMP1-EVs indicated a diameter of around 150 nm. Western blot assay demonstrated that the autophagy related protein LC3-II and p62 are co-secreted with VMP1 in cellular stress models. Furthermore, VMP1-EVs are taken up by different host cells, suggesting that VMP1-EVs may be able to mediate remote communication between cells. In conclusion, we have demonstrated for the first time that the pancreatitis-associated transmembrane protein VMP1 is unconventionally secreted as a component of extracellular vesicles via a secretory autophagy pathway.
Fil: Tadic, Mariana S.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Renna, Felipe Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Herrera Lopez, Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Orquera, Tamara. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Lopez Mingorance, Fabiana Norma. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Ropolo, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Vaccaro, Maria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Materia
EXTRACELLULAR VESICLES
SECRETORY AUTOPHAGY
VMP1
CELLULAR STRESS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/261589
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injuryTadic, Mariana S.Renna, Felipe JavierHerrera Lopez, MalenaOrquera, TamaraLopez Mingorance, Fabiana NormaRopolo, Alejandro JavierVaccaro, Maria InesEXTRACELLULAR VESICLESSECRETORY AUTOPHAGYVMP1CELLULAR STRESShttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Cellular stress activates various mechanisms, including autophagy and vesicular trafficking, to maintain homeostasis and cope with pathological conditions such as acute pancreatitis. One of these mechanisms involves the unconventional secretion of extracellular vesicles (EVs), lipid bilayer-delimited particles released by almost all cell types into the extracellular medium. Recently, an autophagy-related secretory pathway that releases EVs has been discovered. Our study focuses on Vacuole Membrane Protein 1 (VMP1), an autophagy-related protein implicated in pancreatitis, diabetes, pancreatic cancer, and cellular stress management. VMP1 expression triggers autophagy in mammalian cells through its ubiquitination and its direct binding to BECN1. Here, we demonstrate that VMP1 is secreted to the extracellular medium integrated into the membrane of EVs. Using cell lines expressing different VMP1-tag expression plasmids, we successfully isolated and purified VMP1-containing EVs (VMP1-EVs) from the extracellular medium through ultracentrifugation and immune isolation techniques. The secretion of VMP1-EVs was reduced by mTOR inhibition and in the absence of proteins essential for autophagosome formation, such as ATG5. The secretion of endogenous VMP1 by pancreatic acinar cells was highly induced in experimental models of cell stress, including blocked autophagic flux with Bafilomycin and exposure to supramaximal doses of caerulein. Moreover, immuno-isolation of VMP1-EVs from cell models revealed that VMP1 is a component of EV membranes. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis of immuno-isolated VMP1-EVs indicated a diameter of around 150 nm. Western blot assay demonstrated that the autophagy related protein LC3-II and p62 are co-secreted with VMP1 in cellular stress models. Furthermore, VMP1-EVs are taken up by different host cells, suggesting that VMP1-EVs may be able to mediate remote communication between cells. In conclusion, we have demonstrated for the first time that the pancreatitis-associated transmembrane protein VMP1 is unconventionally secreted as a component of extracellular vesicles via a secretory autophagy pathway.Fil: Tadic, Mariana S.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Renna, Felipe Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Herrera Lopez, Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Orquera, Tamara. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Lopez Mingorance, Fabiana Norma. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Ropolo, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaFil: Vaccaro, Maria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; ArgentinaCold Spring Harbor Laboratory Press2024-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/261589Tadic, Mariana S.; Renna, Felipe Javier; Herrera Lopez, Malena; Orquera, Tamara; Lopez Mingorance, Fabiana Norma; et al.; Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury; Cold Spring Harbor Laboratory Press; bioRxiv; 11-2024; 1-252692-8205CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1101/2024.10.31.615473info:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2024.10.31.615473v1info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:25:43Zoai:ri.conicet.gov.ar:11336/261589instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:25:44.036CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury
title Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury
spellingShingle Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury
Tadic, Mariana S.
EXTRACELLULAR VESICLES
SECRETORY AUTOPHAGY
VMP1
CELLULAR STRESS
title_short Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury
title_full Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury
title_fullStr Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury
title_full_unstemmed Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury
title_sort Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury
dc.creator.none.fl_str_mv Tadic, Mariana S.
Renna, Felipe Javier
Herrera Lopez, Malena
Orquera, Tamara
Lopez Mingorance, Fabiana Norma
Ropolo, Alejandro Javier
Vaccaro, Maria Ines
author Tadic, Mariana S.
author_facet Tadic, Mariana S.
Renna, Felipe Javier
Herrera Lopez, Malena
Orquera, Tamara
Lopez Mingorance, Fabiana Norma
Ropolo, Alejandro Javier
Vaccaro, Maria Ines
author_role author
author2 Renna, Felipe Javier
Herrera Lopez, Malena
Orquera, Tamara
Lopez Mingorance, Fabiana Norma
Ropolo, Alejandro Javier
Vaccaro, Maria Ines
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv EXTRACELLULAR VESICLES
SECRETORY AUTOPHAGY
VMP1
CELLULAR STRESS
topic EXTRACELLULAR VESICLES
SECRETORY AUTOPHAGY
VMP1
CELLULAR STRESS
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Cellular stress activates various mechanisms, including autophagy and vesicular trafficking, to maintain homeostasis and cope with pathological conditions such as acute pancreatitis. One of these mechanisms involves the unconventional secretion of extracellular vesicles (EVs), lipid bilayer-delimited particles released by almost all cell types into the extracellular medium. Recently, an autophagy-related secretory pathway that releases EVs has been discovered. Our study focuses on Vacuole Membrane Protein 1 (VMP1), an autophagy-related protein implicated in pancreatitis, diabetes, pancreatic cancer, and cellular stress management. VMP1 expression triggers autophagy in mammalian cells through its ubiquitination and its direct binding to BECN1. Here, we demonstrate that VMP1 is secreted to the extracellular medium integrated into the membrane of EVs. Using cell lines expressing different VMP1-tag expression plasmids, we successfully isolated and purified VMP1-containing EVs (VMP1-EVs) from the extracellular medium through ultracentrifugation and immune isolation techniques. The secretion of VMP1-EVs was reduced by mTOR inhibition and in the absence of proteins essential for autophagosome formation, such as ATG5. The secretion of endogenous VMP1 by pancreatic acinar cells was highly induced in experimental models of cell stress, including blocked autophagic flux with Bafilomycin and exposure to supramaximal doses of caerulein. Moreover, immuno-isolation of VMP1-EVs from cell models revealed that VMP1 is a component of EV membranes. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis of immuno-isolated VMP1-EVs indicated a diameter of around 150 nm. Western blot assay demonstrated that the autophagy related protein LC3-II and p62 are co-secreted with VMP1 in cellular stress models. Furthermore, VMP1-EVs are taken up by different host cells, suggesting that VMP1-EVs may be able to mediate remote communication between cells. In conclusion, we have demonstrated for the first time that the pancreatitis-associated transmembrane protein VMP1 is unconventionally secreted as a component of extracellular vesicles via a secretory autophagy pathway.
Fil: Tadic, Mariana S.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Renna, Felipe Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Herrera Lopez, Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Orquera, Tamara. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Lopez Mingorance, Fabiana Norma. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Ropolo, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
Fil: Vaccaro, Maria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Bioquímica y Medicina Molecular. Universidad de Buenos Aires. Facultad Medicina. Instituto de Bioquímica y Medicina Molecular; Argentina
description Cellular stress activates various mechanisms, including autophagy and vesicular trafficking, to maintain homeostasis and cope with pathological conditions such as acute pancreatitis. One of these mechanisms involves the unconventional secretion of extracellular vesicles (EVs), lipid bilayer-delimited particles released by almost all cell types into the extracellular medium. Recently, an autophagy-related secretory pathway that releases EVs has been discovered. Our study focuses on Vacuole Membrane Protein 1 (VMP1), an autophagy-related protein implicated in pancreatitis, diabetes, pancreatic cancer, and cellular stress management. VMP1 expression triggers autophagy in mammalian cells through its ubiquitination and its direct binding to BECN1. Here, we demonstrate that VMP1 is secreted to the extracellular medium integrated into the membrane of EVs. Using cell lines expressing different VMP1-tag expression plasmids, we successfully isolated and purified VMP1-containing EVs (VMP1-EVs) from the extracellular medium through ultracentrifugation and immune isolation techniques. The secretion of VMP1-EVs was reduced by mTOR inhibition and in the absence of proteins essential for autophagosome formation, such as ATG5. The secretion of endogenous VMP1 by pancreatic acinar cells was highly induced in experimental models of cell stress, including blocked autophagic flux with Bafilomycin and exposure to supramaximal doses of caerulein. Moreover, immuno-isolation of VMP1-EVs from cell models revealed that VMP1 is a component of EV membranes. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis of immuno-isolated VMP1-EVs indicated a diameter of around 150 nm. Western blot assay demonstrated that the autophagy related protein LC3-II and p62 are co-secreted with VMP1 in cellular stress models. Furthermore, VMP1-EVs are taken up by different host cells, suggesting that VMP1-EVs may be able to mediate remote communication between cells. In conclusion, we have demonstrated for the first time that the pancreatitis-associated transmembrane protein VMP1 is unconventionally secreted as a component of extracellular vesicles via a secretory autophagy pathway.
publishDate 2024
dc.date.none.fl_str_mv 2024-11
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/261589
Tadic, Mariana S.; Renna, Felipe Javier; Herrera Lopez, Malena; Orquera, Tamara; Lopez Mingorance, Fabiana Norma; et al.; Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury; Cold Spring Harbor Laboratory Press; bioRxiv; 11-2024; 1-25
2692-8205
CONICET Digital
CONICET
url http://hdl.handle.net/11336/261589
identifier_str_mv Tadic, Mariana S.; Renna, Felipe Javier; Herrera Lopez, Malena; Orquera, Tamara; Lopez Mingorance, Fabiana Norma; et al.; Unconventional secretion of the autophagy-related protein VMP1 via extracellular vesicles as a potential biomarker for pancreatic cell injury; Cold Spring Harbor Laboratory Press; bioRxiv; 11-2024; 1-25
2692-8205
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1101/2024.10.31.615473
info:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2024.10.31.615473v1
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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