Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticles

Surface-exposed calreticulin (CRT) serves as a crucial cell damage-associated molecular pattern for immunogenic apoptosis, by generating an “eat me” signal to macrophages. Aiming at precision immunotherapies we intended to artificially label tumoral cells in vivo with a recombinant CRT, in a targete...

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Detalles Bibliográficos
Autores: Parladé Molist, Eloi|||0000-0001-5750-550X, García León, Annabel, Voltà Durán, Eric|||0000-0003-0017-8274, Unzueta, Ugutz, Mangues, Ramón, Casanova Rigat, Isolda, Villaverde Corrales, Antonio, Vázquez Gómez, Esther
Tipo de recurso: artículo
Fecha de publicación:2024
País:España
Institución:Universitat Politècnica de Catalunya (UPC)
Repositorio:UPCommons. Portal del coneixement obert de la UPC
Idioma:inglés
OAI Identifier:oai:upcommons.upc.edu:2117/452229
Acceso en línea:https://hdl.handle.net/2117/452229
https://dx.doi.org/10.1016/j.ejpb.2024.114410
Access Level:acceso abierto
Palabra clave:Recombinant proteins
Protein materials
Nanoparticles
Drug delivery
Cell targeting
Àrees temàtiques de la UPC::Ciències de la salut::Medicina
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spelling Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticlesParladé Molist, Eloi|||0000-0001-5750-550XGarcía León, AnnabelVoltà Durán, Eric|||0000-0003-0017-8274Unzueta, UgutzMangues, RamónCasanova Rigat, IsoldaVillaverde Corrales, AntonioVázquez Gómez, EstherRecombinant proteinsProtein materialsNanoparticlesDrug deliveryCell targetingÀrees temàtiques de la UPC::Ciències de la salut::MedicinaSurface-exposed calreticulin (CRT) serves as a crucial cell damage-associated molecular pattern for immunogenic apoptosis, by generating an “eat me” signal to macrophages. Aiming at precision immunotherapies we intended to artificially label tumoral cells in vivo with a recombinant CRT, in a targeted way. For that, we have constructed a CRT fusion protein intended to surface attach CXCR4+ cancer cells, to stimulate their immunological destruction. As a targeting ligand of the CRT construct and to drive its specific cell adhesion, we used the peptide V1, a derivative of the vMIP-II cytokine and an antagonist of CXCR4. The modular protein tends to self-assemble as regular 16 nm nanoparticles, assisted by ionic Zn. Through both in vivo and in vitro experiments, we have determined that CRT itself confers cell targeting capabilities to the construct overcoming those of V1, that are only moderate. In particular, CRT binds HeLa cells in absence of further internalization, by a route fully independent of CXCR4. Furthermore, by cytometry in THP-1 cells, we observed that the binding of the protein is preferential for dead cells over live cells, a fact that cannot be associated to a mere artefactual adsorption. These data are discussed in the context of the oligomerizing properties of CRT and the potential clinical applicability of proteins and protein materials functionalized with this novel cell surface ligand.The authors appreciate the financial support received for the development of multimeric recombinant drugs, from AEI (PID2022-136845OB-I00 financed by MCIN/AEI/10.13039/501100011033 to E.V. & A.V. and PID2020-116174RB-I00 to A.V.), AGAUR (SGR2021-00092 to A.V. SGR-CatM.2021-01140 to R.M.), Instituto de Salud Carlos III (PI20/00400 and PI23/00318 to U.U.) co-funded by European Regional Development Fund (ERDF, a way to make Europe). We also appreciate the funding from the CIBER –Consorcio Centro de Investigación Biomédica en Red– (CB06/01/0014 and CB06/01/1031) through TRACC and TRAILS intramural projects (to E.P.), Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación, through the project PI21/00150 to R.M. A.V. received an ICREA ACADEMIA award. The authors are indebted to the Nanotoxicology Unit of the ICTS-141007 Nanbiosis Platform (http://www.nanbiosis.es/portfolio/u18-nanotoxicology-unit/) for the in vivo work. Protein production was partially performed by the ICTS “NANBIOSIS”, more specifically by the Protein Production Platform of CIBER in Bioengineering, Biomaterials & Nanomedicine (CIBER-BBN)/ IBB, at the UAB (http://www.nanbiosis.es/portfolio/u1-protein-production-platform-ppp/). E.V.D was supported by a predoctoral fellowship from Ministerio de Ciencia, Innovación y Universidades (FPU18/04615). UU was supported by Miguel Servet contract (CP19/00028) from ISCIII co-funded by European Social Fund (ESF investing in your future). Cell culture and cytometry experiments were performed at SCAC facilities in the UAB. Confocal microscopy studies were performed at the Servei de Microscòpia in the UAB.Peer Reviewed20242024-09-0120262026-02-02journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/2117/452229https://dx.doi.org/10.1016/j.ejpb.2024.114410reponame:UPCommons. Portal del coneixement obert de la UPCinstname:Universitat Politècnica de Catalunya (UPC)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2http://creativecommons.org/licenses/by-nc/4.0/info:eu-repo/semantics/openAccessoai:upcommons.upc.edu:2117/4522292026-05-27T15:37:01Z
dc.title.none.fl_str_mv Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticles
title Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticles
spellingShingle Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticles
Parladé Molist, Eloi|||0000-0001-5750-550X
Recombinant proteins
Protein materials
Nanoparticles
Drug delivery
Cell targeting
Àrees temàtiques de la UPC::Ciències de la salut::Medicina
title_short Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticles
title_full Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticles
title_fullStr Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticles
title_full_unstemmed Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticles
title_sort Paradoxical cell targeting of calreticulin-empowered, protein-only nanoparticles
dc.creator.none.fl_str_mv Parladé Molist, Eloi|||0000-0001-5750-550X
García León, Annabel
Voltà Durán, Eric|||0000-0003-0017-8274
Unzueta, Ugutz
Mangues, Ramón
Casanova Rigat, Isolda
Villaverde Corrales, Antonio
Vázquez Gómez, Esther
author Parladé Molist, Eloi|||0000-0001-5750-550X
author_facet Parladé Molist, Eloi|||0000-0001-5750-550X
García León, Annabel
Voltà Durán, Eric|||0000-0003-0017-8274
Unzueta, Ugutz
Mangues, Ramón
Casanova Rigat, Isolda
Villaverde Corrales, Antonio
Vázquez Gómez, Esther
author_role author
author2 García León, Annabel
Voltà Durán, Eric|||0000-0003-0017-8274
Unzueta, Ugutz
Mangues, Ramón
Casanova Rigat, Isolda
Villaverde Corrales, Antonio
Vázquez Gómez, Esther
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Recombinant proteins
Protein materials
Nanoparticles
Drug delivery
Cell targeting
Àrees temàtiques de la UPC::Ciències de la salut::Medicina
topic Recombinant proteins
Protein materials
Nanoparticles
Drug delivery
Cell targeting
Àrees temàtiques de la UPC::Ciències de la salut::Medicina
description Surface-exposed calreticulin (CRT) serves as a crucial cell damage-associated molecular pattern for immunogenic apoptosis, by generating an “eat me” signal to macrophages. Aiming at precision immunotherapies we intended to artificially label tumoral cells in vivo with a recombinant CRT, in a targeted way. For that, we have constructed a CRT fusion protein intended to surface attach CXCR4+ cancer cells, to stimulate their immunological destruction. As a targeting ligand of the CRT construct and to drive its specific cell adhesion, we used the peptide V1, a derivative of the vMIP-II cytokine and an antagonist of CXCR4. The modular protein tends to self-assemble as regular 16 nm nanoparticles, assisted by ionic Zn. Through both in vivo and in vitro experiments, we have determined that CRT itself confers cell targeting capabilities to the construct overcoming those of V1, that are only moderate. In particular, CRT binds HeLa cells in absence of further internalization, by a route fully independent of CXCR4. Furthermore, by cytometry in THP-1 cells, we observed that the binding of the protein is preferential for dead cells over live cells, a fact that cannot be associated to a mere artefactual adsorption. These data are discussed in the context of the oligomerizing properties of CRT and the potential clinical applicability of proteins and protein materials functionalized with this novel cell surface ligand.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024-09-01
2026
2026-02-02
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/2117/452229
https://dx.doi.org/10.1016/j.ejpb.2024.114410
url https://hdl.handle.net/2117/452229
https://dx.doi.org/10.1016/j.ejpb.2024.114410
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2

http://creativecommons.org/licenses/by-nc/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2

http://creativecommons.org/licenses/by-nc/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:UPCommons. Portal del coneixement obert de la UPC
instname:Universitat Politècnica de Catalunya (UPC)
instname_str Universitat Politècnica de Catalunya (UPC)
reponame_str UPCommons. Portal del coneixement obert de la UPC
collection UPCommons. Portal del coneixement obert de la UPC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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