Development of Glypican-2 Targeting Single-Domain Antibody CAR T Cells for Neuroblastoma.
Adoptive T-cell therapy
Glypican
Lentivirus
Nanobody
Neuroblastoma
Single-domain antibody
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
14
2
2022
pubmed:
15
2
2022
medline:
19
2
2022
Statut:
ppublish
Résumé
Chimeric antigen receptors (CARs) are engineered fusion proteins constructed from antigen-recognition, signaling, and costimulatory domains. CARs can be expressed in T cells with the purpose of reprogramming the T cells to specifically target tumor cells. This strategy thereby avoids the requirement for antigen processing and presentation by the target cell. Glypican-2 (GPC2) is a cell surface heparan sulfate proteoglycan with highly tumor-specific expression in neuroblastoma compared with nonmalignant cells. Therefore, GPC2 is an attractive target candidate for CAR T-cell therapy. Single-domain antibodies (sdAbs) can access epitopes different from those targeted by single-chain variable fragments and, because of their stability and modularity, could serve as ideal antigen-recognition domains in CAR T cells. Here, we describe a protocol for generating GPC2-targeted sdAb CAR T cells. We also present a methodology for assessing the efficiency of CAR expression on human T cells and their ability to kill GPC2-positive neuroblastoma cells in vitro and in vivo. The method described here is applicable to the production of CAR T cells derived from all types of sdAbs including VHHs and VNARs.
Identifiants
pubmed: 35157288
doi: 10.1007/978-1-0716-2075-5_23
doi:
Substances chimiques
Glypicans
0
Receptors, Antigen, T-Cell
0
Single-Domain Antibodies
0
Types de publication
Journal Article
Research Support, N.I.H., Intramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
451-468Subventions
Organisme : Intramural NIH HHS
ID : Z01 BC010891
Pays : United States
Organisme : Intramural NIH HHS
ID : ZIA BC010891
Pays : United States
Informations de copyright
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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