1. Rational development and characterization of humanized anti-EGFR variant III chimeric antigen receptor T cells for glioblastoma
- Author
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John Scholler, Joseph A. Fraietta, Shannon E. McGettigan, Avery D. Posey, Neeraja Idamakanti, Boris Engels, Prachi R. Patel, Andreas Loew, Christopher C. Kloss, Reshma Singh, Tucker Ezell, Akemi Kosaka, Hideho Okada, Taylor Chen, Alexandria P. Cogdill, Tzvete Dentchev, Arben Nace, Alina C. Boesteanu, Melissa Ramones, John T. Seykora, Jennifer Brogdon, Marcela V. Maus, Laura A. Johnson, Carl H. June, Na Li, Pramod Thekkat, Takayuki Ohkuri, Gabriela Plesa, and Li Zhou
- Subjects
medicine.medical_treatment ,Receptors, Antigen, T-Cell ,Biology ,Medical and Health Sciences ,Vaccine Related ,Mice ,Rare Diseases ,Antigen ,In vivo ,Clinical Research ,Receptors ,medicine ,Genetics ,Cytotoxic T cell ,Animals ,Humans ,Epidermal growth factor receptor ,B cell ,Cancer ,5.2 Cellular and gene therapies ,Brain Neoplasms ,Animal ,General Medicine ,Immunotherapy ,Gene Therapy ,Biological Sciences ,T-Cell ,Molecular biology ,Chimeric antigen receptor ,In vitro ,Human Fetal Tissue ,Brain Disorders ,ErbB Receptors ,Brain Cancer ,Disease Models, Animal ,medicine.anatomical_structure ,Disease Models ,biology.protein ,Heterografts ,Immunization ,Development of treatments and therapeutic interventions ,Glioblastoma ,Biotechnology - 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).
- Published
- 2015