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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/2562
Ver los metadatos del registro completo
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oai:ri.conicet.gov.ar:11336/2562 |
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3498 |
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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 |
_version_ |
1844614245067522048 |
score |
13.070432 |