Pavlina Chuntova1, Yafei Hou1, Ryosuke Naka1,2, Akane Yamamichi1, Tiffany Chen1, Yitzhar Goretsky1, Ryusuke Hatae1, Takahide Nejo1, Gary Kohanbash1,3, Abigail L Mende1, Megan Montoya1, Kira M Downey1, David Diebold1, Jayne Skinner1, Hong-Erh Liang4, Bjoern Schwer1,5,6,7, Hideho Okada1,5,8. 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.
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.
Authors: Luca Gattinoni; Steven E Finkelstein; Christopher A Klebanoff; Paul A Antony; Douglas C Palmer; Paul J Spiess; Leroy N Hwang; Zhiya Yu; Claudia Wrzesinski; David M Heimann; Charles D Surh; Steven A Rosenberg; Nicholas P Restifo Journal: J Exp Med Date: 2005-10-03 Impact factor: 14.307
Authors: Carmen S M Yong; Liza B John; Christel Devaud; Miles H Prince; Ricky W Johnstone; Joseph A Trapani; Phillip K Darcy; Michael H Kershaw Journal: Oncotarget Date: 2016-06-07
Authors: Carter M Suryadevara; Rupen Desai; Melissa L Abel; Katherine A Riccione; Kristen A Batich; Steven H Shen; Pakawat Chongsathidkiet; Patrick C Gedeon; Aladine A Elsamadicy; David J Snyder; James E Herndon; Patrick Healy; Gary E Archer; Bryan D Choi; Peter E Fecci; John H Sampson; Luis Sanchez-Perez Journal: Oncoimmunology Date: 2018-02-21 Impact factor: 8.110