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Preclinical Studies of Granulysin-Based Anti-MUC1-Tn Immunotoxins as a New Antitumoral Treatment

Two granulysin (GRNLY) based immunotoxins were generated, one containing the scFv of the SM3 mAb (SM3GRNLY) and the other the scFv of the AR20.5 mAb (AR20.5GRNLY). These mAb recognize different amino acid sequences of aberrantly O-glycosylated MUC1, also known as the Tn antigen, expressed in a varie...

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Autores principales: Guerrero-Ochoa, Patricia, Ibáñez-Pérez, Raquel, Berbegal-Pinilla, Germán, Aguilar, Diederich, Marzo, Isabel, Corzana, Francisco, Minjárez-Sáenz, Martha, Macías-León, Javier, Conde, Blanca, Raso, Javier, Hurtado-Guerrero, Ramón, Anel, Alberto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9219680/
https://www.ncbi.nlm.nih.gov/pubmed/35740244
http://dx.doi.org/10.3390/biomedicines10061223
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author Guerrero-Ochoa, Patricia
Ibáñez-Pérez, Raquel
Berbegal-Pinilla, Germán
Aguilar, Diederich
Marzo, Isabel
Corzana, Francisco
Minjárez-Sáenz, Martha
Macías-León, Javier
Conde, Blanca
Raso, Javier
Hurtado-Guerrero, Ramón
Anel, Alberto
author_facet Guerrero-Ochoa, Patricia
Ibáñez-Pérez, Raquel
Berbegal-Pinilla, Germán
Aguilar, Diederich
Marzo, Isabel
Corzana, Francisco
Minjárez-Sáenz, Martha
Macías-León, Javier
Conde, Blanca
Raso, Javier
Hurtado-Guerrero, Ramón
Anel, Alberto
author_sort Guerrero-Ochoa, Patricia
collection PubMed
description Two granulysin (GRNLY) based immunotoxins were generated, one containing the scFv of the SM3 mAb (SM3GRNLY) and the other the scFv of the AR20.5 mAb (AR20.5GRNLY). These mAb recognize different amino acid sequences of aberrantly O-glycosylated MUC1, also known as the Tn antigen, expressed in a variety of tumor cell types. We first demonstrated the affinity of these immunotoxins for their antigen using surface plasmon resonance for the purified antigen and flow cytometry for the antigen expressed on the surface of living tumor cells. The induction of cell death of tumor cell lines of different origin positive for Tn antigen expression was stronger in the cases of the immunotoxins than that induced by GRNLY alone. The mechanism of cell death induced by the immunotoxins was studied, showing that the apoptotic component demonstrated previously for GRNLY was also present, but that cell death induced by the immunotoxins included also necroptotic and necrotic components. Finally, we demonstrated the in vivo tumor targeting by the immunotoxins after systemic injection using a xenograft model of the human pancreatic adenocarcinoma CAPAN-2 in athymic mice. While GRNLY alone did not have a therapeutic effect, SM3GRNLY and AR20.5GRNLY reduced tumor volume by 42 and 60%, respectively, compared with untreated tumor-bearing mice, although the results were not statistically significant in the case of AR20.5GRNLY. Histological studies of tumors obtained from treated mice demonstrated reduced cellularity, nuclear morphology compatible with apoptosis induction and active caspase-3 detection by immunohistochemistry. Overall, our results exemplify that these immunotoxins are potential drugs to treat Tn-expressing cancers.
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spelling pubmed-92196802022-06-24 Preclinical Studies of Granulysin-Based Anti-MUC1-Tn Immunotoxins as a New Antitumoral Treatment Guerrero-Ochoa, Patricia Ibáñez-Pérez, Raquel Berbegal-Pinilla, Germán Aguilar, Diederich Marzo, Isabel Corzana, Francisco Minjárez-Sáenz, Martha Macías-León, Javier Conde, Blanca Raso, Javier Hurtado-Guerrero, Ramón Anel, Alberto Biomedicines Article Two granulysin (GRNLY) based immunotoxins were generated, one containing the scFv of the SM3 mAb (SM3GRNLY) and the other the scFv of the AR20.5 mAb (AR20.5GRNLY). These mAb recognize different amino acid sequences of aberrantly O-glycosylated MUC1, also known as the Tn antigen, expressed in a variety of tumor cell types. We first demonstrated the affinity of these immunotoxins for their antigen using surface plasmon resonance for the purified antigen and flow cytometry for the antigen expressed on the surface of living tumor cells. The induction of cell death of tumor cell lines of different origin positive for Tn antigen expression was stronger in the cases of the immunotoxins than that induced by GRNLY alone. The mechanism of cell death induced by the immunotoxins was studied, showing that the apoptotic component demonstrated previously for GRNLY was also present, but that cell death induced by the immunotoxins included also necroptotic and necrotic components. Finally, we demonstrated the in vivo tumor targeting by the immunotoxins after systemic injection using a xenograft model of the human pancreatic adenocarcinoma CAPAN-2 in athymic mice. While GRNLY alone did not have a therapeutic effect, SM3GRNLY and AR20.5GRNLY reduced tumor volume by 42 and 60%, respectively, compared with untreated tumor-bearing mice, although the results were not statistically significant in the case of AR20.5GRNLY. Histological studies of tumors obtained from treated mice demonstrated reduced cellularity, nuclear morphology compatible with apoptosis induction and active caspase-3 detection by immunohistochemistry. Overall, our results exemplify that these immunotoxins are potential drugs to treat Tn-expressing cancers. MDPI 2022-05-24 /pmc/articles/PMC9219680/ /pubmed/35740244 http://dx.doi.org/10.3390/biomedicines10061223 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Guerrero-Ochoa, Patricia
Ibáñez-Pérez, Raquel
Berbegal-Pinilla, Germán
Aguilar, Diederich
Marzo, Isabel
Corzana, Francisco
Minjárez-Sáenz, Martha
Macías-León, Javier
Conde, Blanca
Raso, Javier
Hurtado-Guerrero, Ramón
Anel, Alberto
Preclinical Studies of Granulysin-Based Anti-MUC1-Tn Immunotoxins as a New Antitumoral Treatment
title Preclinical Studies of Granulysin-Based Anti-MUC1-Tn Immunotoxins as a New Antitumoral Treatment
title_full Preclinical Studies of Granulysin-Based Anti-MUC1-Tn Immunotoxins as a New Antitumoral Treatment
title_fullStr Preclinical Studies of Granulysin-Based Anti-MUC1-Tn Immunotoxins as a New Antitumoral Treatment
title_full_unstemmed Preclinical Studies of Granulysin-Based Anti-MUC1-Tn Immunotoxins as a New Antitumoral Treatment
title_short Preclinical Studies of Granulysin-Based Anti-MUC1-Tn Immunotoxins as a New Antitumoral Treatment
title_sort preclinical studies of granulysin-based anti-muc1-tn immunotoxins as a new antitumoral treatment
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9219680/
https://www.ncbi.nlm.nih.gov/pubmed/35740244
http://dx.doi.org/10.3390/biomedicines10061223
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