title: 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.affiliation: Abramson 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.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Johnson
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: John
authors.initials: J
authors.lastname: Scholler
authors.affiliation: Department of Neurosurgery, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ohkuri
authors.affiliation: Department of Neurosurgery, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Akemi
authors.initials: A
authors.lastname: Kosaka
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Prachi R
authors.initials: PR
authors.lastname: Patel
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Shannon E
authors.initials: SE
authors.lastname: McGettigan
authors.affiliation: Department of Dermatology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Arben K
authors.initials: AK
authors.lastname: Nace
authors.affiliation: Department of Dermatology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Tzvete
authors.initials: T
authors.lastname: Dentchev
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Pramod
authors.initials: P
authors.lastname: Thekkat
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Andreas
authors.initials: A
authors.lastname: Loew
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Alina C
authors.initials: AC
authors.lastname: Boesteanu
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Alexandria P
authors.initials: AP
authors.lastname: Cogdill
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Taylor
authors.initials: T
authors.lastname: Chen
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Fraietta
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Kloss
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Avery D
authors.initials: AD
authors.lastname: Posey
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Boris
authors.initials: B
authors.lastname: Engels
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Reshma
authors.initials: R
authors.lastname: Singh
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Tucker
authors.initials: T
authors.lastname: Ezell
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Neeraja
authors.initials: N
authors.lastname: Idamakanti
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Melissa H
authors.initials: MH
authors.lastname: Ramones
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Na
authors.initials: N
authors.lastname: Li
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Plesa
authors.affiliation: Department of Dermatology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: John T
authors.initials: JT
authors.lastname: Seykora
authors.affiliation: Department of Neurosurgery, University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Hideho
authors.initials: H
authors.lastname: Okada
authors.affiliation: Abramson 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.
authors.forename: Carl H
authors.initials: CH
authors.lastname: June
authors.affiliation: Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Brogdon
authors.affiliation: Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA 19104, USA. Department of Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA. marcela.maus@uphs.upenn.edu.
authors.forename: Marcela V
authors.initials: MV
authors.lastname: Maus
mesh_terms: D000818:Animals; D001932:Brain Neoplasms / Q000628:therapy*; D004195:Disease Models, Animal; D066246:ErbB Receptors / Q000276:immunology*; D005909:Glioblastoma / Q000628:therapy*; D064593:Heterografts; D006801:Humans; D007167:Immunotherapy; D051379:Mice; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*
chemical_list: D011948:Receptors, Antigen, T-Cell; D066246:ErbB Receptors