Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1

Autores
Daniels Wells, Tracy R.; Helguera, Gustavo Fernando; Rodríguez, José A.; Leoh, Lai Sum; Erb, Michael A.; Diamante, Graciel; Casero, David; Pellegrini, Mateo; Martinez Maza, Otoniel; Penichet, Manuel L.
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We previously developed an antibody-avidin fusion protein (ch128.1Av) that targets the human transferrin receptor 1 (TfR1) and exhibits direct cytotoxicity against malignant B cells in an iron-dependent manner. ch128.1Av is also a delivery system and its conjugation with biotinylated saporin (b-SO6), a plant ribosome-inactivating toxin, results in a dramatic iron-independent cytotoxicity, both in malignant cells that are sensitive or resistant to ch128.1Av alone, in which the toxin effectively inhibits protein synthesis and triggers caspase activation. We have now found that the ch128.1Av/b-SO6 complex induces a transcriptional response consistent with oxidative stress and DNA damage, a response that is not observed with ch128.1Av alone. Furthermore, we show that the antioxidant N-acetylcysteine partially blocks saporin-induced apoptosis suggesting that oxidative stress contributes to DNA damage and ultimately saporin-induced cell death. Interestingly, the toxin was detected in nuclear extracts by immunoblotting, suggesting the possibility that saporin might induce direct DNA damage. However, confocal microscopy did not show a clear and consistent pattern of intranuclear localization. Finally, using the long-term culture-initiating cell assay we found that ch128.1Av/b-SO6 is not toxic to normal human hematopoietic stem cells suggesting that this critical cell population would be preserved in therapeutic interventions using this immunotoxin.
Fil: Daniels Wells, Tracy R.. University of California; Estados Unidos de América;
Fil: Helguera, Gustavo Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. University of California; Estados Unidos de América;
Fil: Rodríguez, José A.. University of California; Estados Unidos de América;
Fil: Leoh, Lai Sum. University of California; Estados Unidos de América;
Fil: Erb, Michael A.. University of California; Estados Unidos de América;
Fil: Diamante, Graciel. University of California; Estados Unidos de América;
Fil: Casero, David. University Of California; Estados Unidos de América;
Fil: Pellegrini, Mateo. University of California; Estados Unidos de América;
Fil: Martinez Maza, Otoniel. University of California; Estados Unidos de América;
Fil: Penichet, Manuel L.. University of California; Estados Unidos de América;
Materia
Saporin
Immunotoxin
Transferrin Receptor 1
Oxidative Stress
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/2562

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oai_identifier_str oai:ri.conicet.gov.ar:11336/2562
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1Daniels Wells, Tracy R.Helguera, Gustavo FernandoRodríguez, José A.Leoh, Lai SumErb, Michael A.Diamante, GracielCasero, DavidPellegrini, MateoMartinez Maza, OtonielPenichet, Manuel L.SaporinImmunotoxinTransferrin Receptor 1Oxidative Stresshttps://purl.org/becyt/ford/3.4https://purl.org/becyt/ford/3We previously developed an antibody-avidin fusion protein (ch128.1Av) that targets the human transferrin receptor 1 (TfR1) and exhibits direct cytotoxicity against malignant B cells in an iron-dependent manner. ch128.1Av is also a delivery system and its conjugation with biotinylated saporin (b-SO6), a plant ribosome-inactivating toxin, results in a dramatic iron-independent cytotoxicity, both in malignant cells that are sensitive or resistant to ch128.1Av alone, in which the toxin effectively inhibits protein synthesis and triggers caspase activation. We have now found that the ch128.1Av/b-SO6 complex induces a transcriptional response consistent with oxidative stress and DNA damage, a response that is not observed with ch128.1Av alone. Furthermore, we show that the antioxidant N-acetylcysteine partially blocks saporin-induced apoptosis suggesting that oxidative stress contributes to DNA damage and ultimately saporin-induced cell death. Interestingly, the toxin was detected in nuclear extracts by immunoblotting, suggesting the possibility that saporin might induce direct DNA damage. However, confocal microscopy did not show a clear and consistent pattern of intranuclear localization. Finally, using the long-term culture-initiating cell assay we found that ch128.1Av/b-SO6 is not toxic to normal human hematopoietic stem cells suggesting that this critical cell population would be preserved in therapeutic interventions using this immunotoxin.Fil: Daniels Wells, Tracy R.. University of California; Estados Unidos de América;Fil: Helguera, Gustavo Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. University of California; Estados Unidos de América;Fil: Rodríguez, José A.. University of California; Estados Unidos de América;Fil: Leoh, Lai Sum. University of California; Estados Unidos de América;Fil: Erb, Michael A.. University of California; Estados Unidos de América;Fil: Diamante, Graciel. University of California; Estados Unidos de América;Fil: Casero, David. University Of California; Estados Unidos de América;Fil: Pellegrini, Mateo. University of California; Estados Unidos de América;Fil: Martinez Maza, Otoniel. University of California; Estados Unidos de América;Fil: Penichet, Manuel L.. University of California; Estados Unidos de América;Pergamon-elsevier Science Ltd2012-10-17info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/2562Daniels Wells, Tracy R.; Helguera, Gustavo Fernando; Rodríguez, José A.; Leoh, Lai Sum; Erb, Michael A.; et al.; Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1; Pergamon-elsevier Science Ltd; Toxicology In Vitro; 27; 1; 17-10-2012; 220-2310887-23331879-3177enginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.tiv.2012.10.006info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0887233312002822info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:24:47Zoai:ri.conicet.gov.ar:11336/2562instacron: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 10:24:47.5CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1
title Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1
spellingShingle Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1
Daniels Wells, Tracy R.
Saporin
Immunotoxin
Transferrin Receptor 1
Oxidative Stress
title_short Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1
title_full Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1
title_fullStr Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1
title_full_unstemmed Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1
title_sort Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1
dc.creator.none.fl_str_mv Daniels Wells, Tracy R.
Helguera, Gustavo Fernando
Rodríguez, José A.
Leoh, Lai Sum
Erb, Michael A.
Diamante, Graciel
Casero, David
Pellegrini, Mateo
Martinez Maza, Otoniel
Penichet, Manuel L.
author Daniels Wells, Tracy R.
author_facet Daniels Wells, Tracy R.
Helguera, Gustavo Fernando
Rodríguez, José A.
Leoh, Lai Sum
Erb, Michael A.
Diamante, Graciel
Casero, David
Pellegrini, Mateo
Martinez Maza, Otoniel
Penichet, Manuel L.
author_role author
author2 Helguera, Gustavo Fernando
Rodríguez, José A.
Leoh, Lai Sum
Erb, Michael A.
Diamante, Graciel
Casero, David
Pellegrini, Mateo
Martinez Maza, Otoniel
Penichet, Manuel L.
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Saporin
Immunotoxin
Transferrin Receptor 1
Oxidative Stress
topic Saporin
Immunotoxin
Transferrin Receptor 1
Oxidative Stress
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.4
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv We previously developed an antibody-avidin fusion protein (ch128.1Av) that targets the human transferrin receptor 1 (TfR1) and exhibits direct cytotoxicity against malignant B cells in an iron-dependent manner. ch128.1Av is also a delivery system and its conjugation with biotinylated saporin (b-SO6), a plant ribosome-inactivating toxin, results in a dramatic iron-independent cytotoxicity, both in malignant cells that are sensitive or resistant to ch128.1Av alone, in which the toxin effectively inhibits protein synthesis and triggers caspase activation. We have now found that the ch128.1Av/b-SO6 complex induces a transcriptional response consistent with oxidative stress and DNA damage, a response that is not observed with ch128.1Av alone. Furthermore, we show that the antioxidant N-acetylcysteine partially blocks saporin-induced apoptosis suggesting that oxidative stress contributes to DNA damage and ultimately saporin-induced cell death. Interestingly, the toxin was detected in nuclear extracts by immunoblotting, suggesting the possibility that saporin might induce direct DNA damage. However, confocal microscopy did not show a clear and consistent pattern of intranuclear localization. Finally, using the long-term culture-initiating cell assay we found that ch128.1Av/b-SO6 is not toxic to normal human hematopoietic stem cells suggesting that this critical cell population would be preserved in therapeutic interventions using this immunotoxin.
Fil: Daniels Wells, Tracy R.. University of California; Estados Unidos de América;
Fil: Helguera, Gustavo Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. University of California; Estados Unidos de América;
Fil: Rodríguez, José A.. University of California; Estados Unidos de América;
Fil: Leoh, Lai Sum. University of California; Estados Unidos de América;
Fil: Erb, Michael A.. University of California; Estados Unidos de América;
Fil: Diamante, Graciel. University of California; Estados Unidos de América;
Fil: Casero, David. University Of California; Estados Unidos de América;
Fil: Pellegrini, Mateo. University of California; Estados Unidos de América;
Fil: Martinez Maza, Otoniel. University of California; Estados Unidos de América;
Fil: Penichet, Manuel L.. University of California; Estados Unidos de América;
description We previously developed an antibody-avidin fusion protein (ch128.1Av) that targets the human transferrin receptor 1 (TfR1) and exhibits direct cytotoxicity against malignant B cells in an iron-dependent manner. ch128.1Av is also a delivery system and its conjugation with biotinylated saporin (b-SO6), a plant ribosome-inactivating toxin, results in a dramatic iron-independent cytotoxicity, both in malignant cells that are sensitive or resistant to ch128.1Av alone, in which the toxin effectively inhibits protein synthesis and triggers caspase activation. We have now found that the ch128.1Av/b-SO6 complex induces a transcriptional response consistent with oxidative stress and DNA damage, a response that is not observed with ch128.1Av alone. Furthermore, we show that the antioxidant N-acetylcysteine partially blocks saporin-induced apoptosis suggesting that oxidative stress contributes to DNA damage and ultimately saporin-induced cell death. Interestingly, the toxin was detected in nuclear extracts by immunoblotting, suggesting the possibility that saporin might induce direct DNA damage. However, confocal microscopy did not show a clear and consistent pattern of intranuclear localization. Finally, using the long-term culture-initiating cell assay we found that ch128.1Av/b-SO6 is not toxic to normal human hematopoietic stem cells suggesting that this critical cell population would be preserved in therapeutic interventions using this immunotoxin.
publishDate 2012
dc.date.none.fl_str_mv 2012-10-17
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/11336/2562
Daniels Wells, Tracy R.; Helguera, Gustavo Fernando; Rodríguez, José A.; Leoh, Lai Sum; Erb, Michael A.; et al.; Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1; Pergamon-elsevier Science Ltd; Toxicology In Vitro; 27; 1; 17-10-2012; 220-231
0887-2333
1879-3177
url http://hdl.handle.net/11336/2562
identifier_str_mv Daniels Wells, Tracy R.; Helguera, Gustavo Fernando; Rodríguez, José A.; Leoh, Lai Sum; Erb, Michael A.; et al.; Insights into the mechanism of cell death induced by saporin delivered into cancer cells by an antibody fusion protein targeting the transferrin receptor 1; Pergamon-elsevier Science Ltd; Toxicology In Vitro; 27; 1; 17-10-2012; 220-231
0887-2333
1879-3177
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.tiv.2012.10.006
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0887233312002822
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Pergamon-elsevier Science Ltd
publisher.none.fl_str_mv Pergamon-elsevier Science Ltd
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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