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Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors

The Notch signaling ligand JAG1 is overexpressed in various aggressive tumors and is associated with poor clinical prognosis. Hence, therapies targeting oncogenic JAG1 hold great potential for the treatment of certain tumors. Here, we report the identification of specific anti-JAG1 single-chain vari...

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Autores principales: Silva, Gabriela, Rodrigues, Ana F., Ferreira, Susana, Matos, Carolina, Eleutério, Rute P., Marques, Gonçalo, Kucheryava, Khrystyna, Lemos, Ana R., Sousa, Pedro M. F., Castro, Rute, Barbas, Ana, Simão, Daniel, Alves, Paula M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10046313/
https://www.ncbi.nlm.nih.gov/pubmed/36979394
http://dx.doi.org/10.3390/biom13030459
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author Silva, Gabriela
Rodrigues, Ana F.
Ferreira, Susana
Matos, Carolina
Eleutério, Rute P.
Marques, Gonçalo
Kucheryava, Khrystyna
Lemos, Ana R.
Sousa, Pedro M. F.
Castro, Rute
Barbas, Ana
Simão, Daniel
Alves, Paula M.
author_facet Silva, Gabriela
Rodrigues, Ana F.
Ferreira, Susana
Matos, Carolina
Eleutério, Rute P.
Marques, Gonçalo
Kucheryava, Khrystyna
Lemos, Ana R.
Sousa, Pedro M. F.
Castro, Rute
Barbas, Ana
Simão, Daniel
Alves, Paula M.
author_sort Silva, Gabriela
collection PubMed
description The Notch signaling ligand JAG1 is overexpressed in various aggressive tumors and is associated with poor clinical prognosis. Hence, therapies targeting oncogenic JAG1 hold great potential for the treatment of certain tumors. Here, we report the identification of specific anti-JAG1 single-chain variable fragments (scFvs), one of them endowing chimeric antigen receptor (CAR) T cells with cytotoxicity against JAG1-positive cells. Anti-JAG1 scFvs were identified from human phage display libraries, reformatted into full-length monoclonal antibodies (Abs), and produced in mammalian cells. The characterization of these Abs identified two specific anti-JAG1 Abs (J1.B5 and J1.F1) with nanomolar affinities. Cloning the respective scFv sequences in our second- and third-generation CAR backbones resulted in six anti-JAG1 CAR constructs, which were screened for JAG1-mediated T-cell activation in Jurkat T cells in coculture assays with JAG1-positive cell lines. Studies in primary T cells demonstrated that one CAR harboring the J1.B5 scFv significantly induced effective T-cell activation in the presence of JAG1-positive, but not in JAG1-knockout, cancer cells, and enabled specific killing of JAG1-positive cells. Thus, this new anti-JAG1 scFv represents a promising candidate for the development of cell therapies against JAG1-positive tumors.
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spelling pubmed-100463132023-03-29 Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors Silva, Gabriela Rodrigues, Ana F. Ferreira, Susana Matos, Carolina Eleutério, Rute P. Marques, Gonçalo Kucheryava, Khrystyna Lemos, Ana R. Sousa, Pedro M. F. Castro, Rute Barbas, Ana Simão, Daniel Alves, Paula M. Biomolecules Article The Notch signaling ligand JAG1 is overexpressed in various aggressive tumors and is associated with poor clinical prognosis. Hence, therapies targeting oncogenic JAG1 hold great potential for the treatment of certain tumors. Here, we report the identification of specific anti-JAG1 single-chain variable fragments (scFvs), one of them endowing chimeric antigen receptor (CAR) T cells with cytotoxicity against JAG1-positive cells. Anti-JAG1 scFvs were identified from human phage display libraries, reformatted into full-length monoclonal antibodies (Abs), and produced in mammalian cells. The characterization of these Abs identified two specific anti-JAG1 Abs (J1.B5 and J1.F1) with nanomolar affinities. Cloning the respective scFv sequences in our second- and third-generation CAR backbones resulted in six anti-JAG1 CAR constructs, which were screened for JAG1-mediated T-cell activation in Jurkat T cells in coculture assays with JAG1-positive cell lines. Studies in primary T cells demonstrated that one CAR harboring the J1.B5 scFv significantly induced effective T-cell activation in the presence of JAG1-positive, but not in JAG1-knockout, cancer cells, and enabled specific killing of JAG1-positive cells. Thus, this new anti-JAG1 scFv represents a promising candidate for the development of cell therapies against JAG1-positive tumors. MDPI 2023-03-02 /pmc/articles/PMC10046313/ /pubmed/36979394 http://dx.doi.org/10.3390/biom13030459 Text en © 2023 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
Silva, Gabriela
Rodrigues, Ana F.
Ferreira, Susana
Matos, Carolina
Eleutério, Rute P.
Marques, Gonçalo
Kucheryava, Khrystyna
Lemos, Ana R.
Sousa, Pedro M. F.
Castro, Rute
Barbas, Ana
Simão, Daniel
Alves, Paula M.
Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors
title Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors
title_full Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors
title_fullStr Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors
title_full_unstemmed Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors
title_short Novel scFv against Notch Ligand JAG1 Suitable for Development of Cell Therapies toward JAG1-Positive Tumors
title_sort novel scfv against notch ligand jag1 suitable for development of cell therapies toward jag1-positive tumors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10046313/
https://www.ncbi.nlm.nih.gov/pubmed/36979394
http://dx.doi.org/10.3390/biom13030459
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