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Updated: Apr 17, 2026
Generation of CAR T Cells for Adoptive Therapy in the Context of Glioblastoma Standard of Care
Published on: February 16, 2015
Rational development and characterization of humanized anti-EGFR variant III chimeric antigen receptor T cells for
Laura A Johnson1, John Scholler2, Takayuki Ohkuri3
1Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
Abstract:
Chimeric antigen receptors (CARs) are synthetic molecules designed to redirect T cells to specific antigens. CAR-modified T cells can mediate long-term durable remissions in B cell malignancies, but expanding this platform to solid tumors requires the discovery of surface targets with limited expression in normal tissues. The variant III mutation of the epidermal growth factor receptor (EGFRvIII) results from an in-frame deletion of a portion of the extracellular domain, creating a neoepitope. We chose a vector backbone encoding a second-generation CAR based on efficacy of a murine scFv-based CAR in a xenograft model of glioblastoma. Next, we generated a panel of humanized scFvs and tested their specificity and function as soluble proteins and in the form of CAR-transduced T cells; a low-affinity scFv was selected on the basis of its specificity for EGFRvIII over wild-type EGFR. The lead candidate scFv was tested in vitro for its ability to direct CAR-transduced T cells to specifically lyse, proliferate, and secrete cytokines in response to antigen-bearing targets. We further evaluated the specificity of the lead CAR candidate in vitro against EGFR-expressing keratinocytes and in vivo in a model of mice grafted with normal human skin. EGFRvIII-directed CAR T cells were also able to control tumor growth in xenogeneic subcutaneous and orthotopic models of human EGFRvIII(+) glioblastoma. On the basis of these results, we have designed a phase 1 clinical study of CAR T cells transduced with humanized scFv directed to EGFRvIII in patients with either residual or recurrent glioblastoma (NCT02209376).
Insights
Chimeric antigen receptor (CAR) T cells targeting the EGFRvIII mutation show promise for treating glioblastoma. These engineered T cells effectively controlled tumor growth in preclinical models, leading to a phase 1 clinical trial.
Area of Science:
- Oncology
- Immunotherapy
- Molecular Biology
Background:
- Chimeric antigen receptor (CAR) T cell therapy has shown success in B cell malignancies.
- Expanding CAR T cell therapy to solid tumors requires identifying specific targets with minimal expression on normal tissues.
- Epidermal growth factor receptor variant III (EGFRvIII) is a tumor-specific neoepitope found in glioblastoma.
Purpose of the Study:
- To develop and evaluate EGFRvIII-targeted CAR T cells for glioblastoma treatment.
- To identify a specific single-chain variable fragment (scFv) that targets EGFRvIII with high specificity.
- To assess the in vitro and in vivo efficacy and safety of EGFRvIII-directed CAR T cells.
Main Methods:
- Generated and screened humanized scFvs for specificity against EGFRvIII over wild-type EGFR.
- Tested lead CAR T cells in vitro for specific lysis, proliferation, and cytokine secretion against antigen-bearing targets.
- Evaluated CAR T cell specificity in vitro against EGFR-expressing keratinocytes and in vivo in mice with normal human skin grafts.
- Assessed tumor growth control in xenogeneic subcutaneous and orthotopic models of human glioblastoma.
Main Results:
- Selected a low-affinity humanized scFv with high specificity for EGFRvIII over wild-type EGFR.
- Demonstrated that EGFRvIII-directed CAR T cells specifically lyse, proliferate, and secrete cytokines in response to antigen-positive targets.
- Confirmed in vitro and in vivo specificity against normal tissues expressing EGFR.
- Showed significant control of glioblastoma tumor growth in both subcutaneous and orthotopic xenograft models.
Conclusions:
- EGFRvIII-directed CAR T cells are a promising therapeutic strategy for glioblastoma.
- Preclinical data support the safety and efficacy of targeting EGFRvIII for glioblastoma treatment.
- A phase 1 clinical trial (NCT02209376) has been initiated to evaluate these CAR T cells in patients with glioblastoma.
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