Novel EGFRvIII-CAR transgenic mice for rigorous preclinical studies in syngeneic mice

Pavlina Chuntova; Yafei Hou; Ryosuke Naka; Akane Yamamichi; Tiffany Chen; Yitzhar Goretsky; Ryusuke Hatae; Takahide Nejo; Gary Kohanbash; Abigail L Mende; Megan Montoya; Kira M Downey; David Diebold; Jayne Skinner; Hong-Erh Liang; Bjoern Schwer; Hideho Okada

doi:10.1093/neuonc/noab182

Novel EGFRvIII-CAR transgenic mice for rigorous preclinical studies in syngeneic mice

Neuro Oncol. 2022 Feb 1;24(2):259-272. doi: 10.1093/neuonc/noab182.

Authors

Pavlina Chuntova¹, Yafei Hou¹, Ryosuke Naka^{1

2}, Akane Yamamichi¹, Tiffany Chen¹, Yitzhar Goretsky¹, Ryusuke Hatae¹, Takahide Nejo¹, Gary Kohanbash^{1

3}, Abigail L Mende¹, Megan Montoya¹, Kira M Downey¹, David Diebold¹, Jayne Skinner¹, Hong-Erh Liang⁴, Bjoern Schwer^{1

5

6

7}, Hideho Okada^{1

5

8}

Affiliations

¹ Department of Neurological Surgery, University of California, San Francisco, San Francisco, California, USA.
² Japanese Red Cross Osaka Hospital, Osaka City, Japan.
³ Department of Neurological Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
⁴ Department of Medicine, University of California, San Francisco, San Francisco, California, USA.
⁵ Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, California, USA.
⁶ The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, California, USA.
⁷ Kavli Institute for Fundamental Neuroscience, University of California, San Francisco, San Francisco, California, USA.
⁸ The Parker Institute for Cancer Immunotherapy, San Francisco, California, USA.

Abstract

Background: Rigorous preclinical studies of chimeric antigen receptor (CAR) immunotherapy will require large quantities of consistent and high-quality CAR-transduced T (CART) cells that can be used in syngeneic mouse glioblastoma (GBM) models. To this end, we developed a novel transgenic (Tg) mouse strain with a fully murinized CAR targeting epidermal growth factor receptor variant III (EGFRvIII).

Methods: We first established the murinized version of EGFRvIII-CAR and validated its function using a retroviral vector (RV) in C57BL/6J mice bearing syngeneic SB28 GBM expressing EGFRvIII. Next, we created C57BL/6J-background Tg mice carrying the anti-EGFRvIII-CAR downstream of a Lox-Stop-Lox cassette in the Rosa26 locus. We bred these mice with CD4-Cre Tg mice to allow CAR expression on T cells and evaluated the function of the CART cells both in vitro and in vivo. To inhibit immunosuppressive myeloid cells within SB28 GBM, we also evaluated a combination approach of CART and an anti-EP4 compound (ONO-AE3-208).

Results: Both RV- and Tg-CART cells demonstrated specific cytotoxic activities against SB28-EGFRvIII cells. A single intravenous infusion of EGFRvIII-CART cells prolonged the survival of glioma-bearing mice when preceded by a lymphodepletion regimen with recurrent tumors displaying profound EGFRvIII loss. The addition of ONO-AE3-208 resulted in long-term survival in a fraction of CART-treated mice and those survivors demonstrated delayed growth of subcutaneously re-challenged both EGFRvIII+ and parental EGFRvIII- SB28.

Conclusion: Our new syngeneic CAR Tg mouse model can serve as a useful tool to address clinically relevant questions and develop future immunotherapeutic strategies.

Keywords: CART cells; EGFRvIII; glioblastoma; immunotherapy; transgenic.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line, Tumor
ErbB Receptors*
Glioblastoma* / pathology
Immunotherapy, Adoptive* / methods
Mice
Mice, Inbred C57BL
Mice, Transgenic
Receptors, Chimeric Antigen*

Substances

Receptors, Chimeric Antigen
epidermal growth factor receptor VIII
ErbB Receptors

Abstract

Publication types

MeSH terms

Substances

Grants and funding