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Pre-clinical validation of a humanized anti-EGFR variant III chimeric antigen receptor and phase I trial of CART-EGFRvIII in glioblastoma

  • Laura A Johnson
  • John Scholler
  • Takayuki Ohkuri
  • Akemi Kosaka
  • Prachi R Patel
  • Shannon E McGettigan
  • Arben Nace
  • Pramod Thekkat
  • Andreas Loew
  • Taylor J Chen
  • Joseph A Fraietta
  • Avery D Posey
  • Alina C Boesteanu
  • Alexandria P Cogdill
  • Boris Engels
  • Reshma Singh
  • Tucker R Ezell
  • Neeraja Idamakanti
  • Gabriela Plesa
  • John Seykora
  • Hideho Okada
  • Carl June
  • Jennifer Brogdon
  • Marcela Maus
Open Access
Oral presentation

Keywords

Epidermal Growth Factor Receptor Chimeric Antigen Receptor Normal Human Skin Control Tumor Growth Lead Candidate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Chimeric antigen receptors are synthetic molecules designed to re-direct T cells to specific surface 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 novel surface targets with limited expression. The variant III mutation of the epidermal growth factor receptor (EGFR variant III) is the most common variant of the EGF receptor observed in human tumors, and results from an in-frame deletion of a portion of the extracellular domain. In glioblastoma, the EGFRvIII mutation is oncogenic, portends a poor prognosis, and is thought to be enriched in glioblastoma stem cells. However, because the neoepitope of EGFR variant III is based on a small peptide sequence, an antibody or single-chain variable fragment (scFv) directed to this epitope must be rigorously tested to confirm lack of cross-reactivity to the ubiquitously expressed normal EGFR. Having selected a candidate murine scFv directed to EGFRvIII and a vector backbone encoding a second generation CAR, we generated a panel of humanized scFv's and tested their specificity and function as soluble proteins and in the form of CAR-transduced T cells. The lead candidate scFv was tested in vitro for its ability to direct CAR-transduced T cells to kill antigen-bearing targets effectively, and proliferate and secrete cytokines specifically in response to antigen. We further evaluated the specificity of the lead candidate CAR by comparing it to a cetuximab-based CAR which does not discriminate between EGFR and EGFR variant III; the two CARs, along with negative controls, were tested in vitro against primary cells derived from a panel of normal tissues, and in vivo in immunodeficient mice grafted with normal human skin, which naturally expresses EGFR. CAR-T cells were also able to control tumor growth in xenogeneic subcutaneous and orthotopic models of human EGFR variant III+ glioblastoma. We have designed a Phase I clinical study of CAR T cells transduced with humanized scFv directed to EGFR variant III in patients with glioblastoma.

Copyright information

© Johnson et al.; licensee BioMed Central Ltd. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Authors and Affiliations

  • Laura A Johnson
    • 1
  • John Scholler
    • 2
  • Takayuki Ohkuri
    • 3
  • Akemi Kosaka
    • 3
  • Prachi R Patel
    • 2
  • Shannon E McGettigan
    • 4
  • Arben Nace
    • 5
  • Pramod Thekkat
    • 6
  • Andreas Loew
    • 7
  • Taylor J Chen
    • 2
  • Joseph A Fraietta
    • 1
  • Avery D Posey
    • 2
  • Alina C Boesteanu
    • 8
  • Alexandria P Cogdill
    • 2
  • Boris Engels
    • 7
  • Reshma Singh
    • 7
  • Tucker R Ezell
    • 7
  • Neeraja Idamakanti
    • 9
  • Gabriela Plesa
    • 10
  • John Seykora
    • 2
  • Hideho Okada
    • 11
  • Carl June
    • 2
  • Jennifer Brogdon
    • 7
  • Marcela Maus
    • 12
  1. 1.Translational Research ProgramUniversity of Pennsylvania Perelman School of MedicinePhiladelphiaUSA
  2. 2.University of PennsylvaniaPhiladelphiaUSA
  3. 3.University of PittsburghPittsburghUSA
  4. 4.University of PennsylvaniaHatboroUSA
  5. 5.University of PennsylvaniaLandenbergUSA
  6. 6.Novartis Institutes of BioMedical Research IncQuincyUSA
  7. 7.Novartis Institutes for Biomedical Research IncCambridgeUSA
  8. 8.University of Pennsylvania Abramson Cancer CenterWilow GroveUSA
  9. 9.NovartisBurlingtonUSA
  10. 10.University of PennsylvaniaBlue BellUSA
  11. 11.University of California San FranciscoSan FranciscoUSA
  12. 12.Abramson Cancer Center, Dept. of MedicineUniversity of Pennsylvania Perelman School of MedicineBryn MawrUSA

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