Rational development and characterization of humanized anti-EGFR variant III chimeric antigen receptor T cells for glioblastoma.

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).

• Authors: Laura A Johnson, John Scholler, Takayuki Ohkuri, Akemi Kosaka, Prachi R Patel, Shannon E McGettigan, Arben K Nace, Tzvete Dentchev, Pramod Thekkat, Andreas Loew, Alina C Boesteanu, Alexandria P Cogdill, Taylor Chen, Joseph A Fraietta, Christopher C Kloss, Avery D Posey Jr, Boris Engels, Reshma Singh, Tucker Ezell, Neeraja Idamakanti, Melissa H Ramones, Na Li, Li Zhou, Gabriela Plesa, John T Seykora, Hideho Okada, Carl H June, Jennifer L Brogdon, Marcela V Maus
• Journal: Science translational medicine (Sci Transl Med) Vol. 7 Issue 275 Pg. 275ra22 (Feb 18 2015) ISSN: 1946-6242 [Electronic] United States
• PMID: 25696001 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015, American Association for the Advancement of Science.

Chemical References

• Receptors, Antigen, T-Cell
• ErbB Receptors

Topics

• Animals
• Brain Neoplasms (therapy)
• Disease Models, Animal
• ErbB Receptors (immunology)
• Glioblastoma (therapy)
• Heterografts
• Humans
• Immunotherapy
• Mice
• Receptors, Antigen, T-Cell (immunology)