Characterization and functional analysis of scFv-based chimeric antigen receptors to redirect T Cells to IL13Rα2-positive glioma

Giedre Krenciute, Simone Krebs, David Torres, Meng Fen Wu, Hao Liu, Gianpietro Dotti, Xiao Nan Li, Maciej S. Lesniak, Irina V. Balyasnikova, Stephen Gottschalk*

*Corresponding author for this work

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Immunotherapy with T cells expressing chimeric antigen receptors (CARs) is an attractive approach to improve outcomes for patients with glioblastoma (GBM). IL13Rα2 is expressed at a high frequency in GBM but not in normal brain, making it a promising CAR T-cell therapy target. IL13Rα2-specific CARs generated up to date contain mutated forms of IL13 as an antigen-binding domain. While these CARs target IL13Rα2, they also recognize IL13Rα1, which is broadly expressed. To overcome this limitation, we constructed a panel of IL13Rα2-specific CARs that contain the IL13Rα2-specific single-chain variable fragment (scFv) 47 as an antigen binding domain, short or long spacer regions, a transmembrane domain, and endodomains derived from costimulatory molecules and CD3.ζ (IL13Rα2-CARs). IL13Rα2-CAR T cells recognized IL13Rα2-positive target cells in coculture and cytotoxicity assays with no cross-reactivity to IL13Rα1. However, only IL13Rα2-CAR T cells with a short spacer region produced IL2 in an antigen-dependent fashion. In vivo, T cells expressing IL13Rα2-CARs with short spacer regions and CD28.ζ, 41BB.ζ, and CD28.OX40.ζ endodomains had potent anti-glioma activity conferring a significant survival advantage in comparison to mice that received control T cells. Thus, IL13Rα2-CAR T cells hold the promise to improve current IL13Rα2-targeted immunotherapy approaches for GBM and other IL13Rα2-positive malignancies.

Original languageEnglish (US)
Pages (from-to)354-363
Number of pages10
JournalMolecular Therapy
Volume24
Issue number2
DOIs
StatePublished - Feb 1 2016

Fingerprint

Antigen Receptors
Glioma
T-Lymphocytes
Glioblastoma
T-Cell Antigen Receptor
Antigens
Immunotherapy
Single-Chain Antibodies
Interleukin-13
Cell- and Tissue-Based Therapy
Coculture Techniques
Interleukin-2
Survival
Brain

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

Cite this

Krenciute, Giedre ; Krebs, Simone ; Torres, David ; Wu, Meng Fen ; Liu, Hao ; Dotti, Gianpietro ; Li, Xiao Nan ; Lesniak, Maciej S. ; Balyasnikova, Irina V. ; Gottschalk, Stephen. / Characterization and functional analysis of scFv-based chimeric antigen receptors to redirect T Cells to IL13Rα2-positive glioma. In: Molecular Therapy. 2016 ; Vol. 24, No. 2. pp. 354-363.
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abstract = "Immunotherapy with T cells expressing chimeric antigen receptors (CARs) is an attractive approach to improve outcomes for patients with glioblastoma (GBM). IL13Rα2 is expressed at a high frequency in GBM but not in normal brain, making it a promising CAR T-cell therapy target. IL13Rα2-specific CARs generated up to date contain mutated forms of IL13 as an antigen-binding domain. While these CARs target IL13Rα2, they also recognize IL13Rα1, which is broadly expressed. To overcome this limitation, we constructed a panel of IL13Rα2-specific CARs that contain the IL13Rα2-specific single-chain variable fragment (scFv) 47 as an antigen binding domain, short or long spacer regions, a transmembrane domain, and endodomains derived from costimulatory molecules and CD3.ζ (IL13Rα2-CARs). IL13Rα2-CAR T cells recognized IL13Rα2-positive target cells in coculture and cytotoxicity assays with no cross-reactivity to IL13Rα1. However, only IL13Rα2-CAR T cells with a short spacer region produced IL2 in an antigen-dependent fashion. In vivo, T cells expressing IL13Rα2-CARs with short spacer regions and CD28.ζ, 41BB.ζ, and CD28.OX40.ζ endodomains had potent anti-glioma activity conferring a significant survival advantage in comparison to mice that received control T cells. Thus, IL13Rα2-CAR T cells hold the promise to improve current IL13Rα2-targeted immunotherapy approaches for GBM and other IL13Rα2-positive malignancies.",
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Characterization and functional analysis of scFv-based chimeric antigen receptors to redirect T Cells to IL13Rα2-positive glioma. / Krenciute, Giedre; Krebs, Simone; Torres, David; Wu, Meng Fen; Liu, Hao; Dotti, Gianpietro; Li, Xiao Nan; Lesniak, Maciej S.; Balyasnikova, Irina V.; Gottschalk, Stephen.

In: Molecular Therapy, Vol. 24, No. 2, 01.02.2016, p. 354-363.

Research output: Contribution to journalArticle

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