GPC2-CAR T cells tuned for low antigen density mediate potent activity against neuroblastoma without toxicity

Sabine Heitzeneder; Kristopher R Bosse; Zhongyu Zhu; Doncho Zhelev; Robbie G Majzner; Molly T Radosevich; Shaurya Dhingra; Elena Sotillo; Samantha Buongervino; Guillem Pascual-Pasto; Emily Garrigan; Peng Xu; Jing Huang; Benjamin Salzer; Alberto Delaidelli; Swetha Raman; Hong Cui; Benjamin Martinez; Scott J Bornheimer; Bita Sahaf; Anya Alag; Irfete S Fetahu; Martin Hasselblatt; Kevin R Parker; Hima Anbunathan; Jennifer Hwang; Min Huang; Kathleen Sakamoto; Norman J Lacayo; Dorota D Klysz; Johanna Theruvath; José G Vilches-Moure; Ansuman T Satpathy; Howard Y Chang; Manfred Lehner; Sabine Taschner-Mandl; Jean-Phillipe Julien; Poul H Sorensen; Dimiter S Dimitrov; John M Maris; Crystal L Mackall

doi:10.1016/j.ccell.2021.12.005

GPC2-CAR T cells tuned for low antigen density mediate potent activity against neuroblastoma without toxicity

Cancer Cell. 2022 Jan 10;40(1):53-69.e9. doi: 10.1016/j.ccell.2021.12.005. Epub 2021 Dec 30.

Authors

Sabine Heitzeneder¹, Kristopher R Bosse², Zhongyu Zhu³, Doncho Zhelev⁴, Robbie G Majzner¹, Molly T Radosevich¹, Shaurya Dhingra¹, Elena Sotillo¹, Samantha Buongervino⁵, Guillem Pascual-Pasto⁵, Emily Garrigan⁵, Peng Xu¹, Jing Huang¹, Benjamin Salzer⁶, Alberto Delaidelli⁷, Swetha Raman⁸, Hong Cui⁸, Benjamin Martinez⁸, Scott J Bornheimer⁹, Bita Sahaf¹, Anya Alag¹, Irfete S Fetahu⁴, Martin Hasselblatt¹⁰, Kevin R Parker¹¹, Hima Anbunathan¹, Jennifer Hwang³, Min Huang¹², Kathleen Sakamoto¹², Norman J Lacayo¹², Dorota D Klysz¹, Johanna Theruvath¹, José G Vilches-Moure¹³, Ansuman T Satpathy¹⁴, Howard Y Chang¹⁵, Manfred Lehner⁶, Sabine Taschner-Mandl¹⁶, Jean-Phillipe Julien¹⁷, Poul H Sorensen⁷, Dimiter S Dimitrov⁴, John M Maris², Crystal L Mackall¹⁸

Affiliations

¹ Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Lorry Lokey Building, Suite G3141, MC: 5456, 265 Campus Drive, Stanford, CA 94305, USA.
² Children's Hospital of Philadelphia and Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA; Department of Pediatrics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
³ National Cancer Institute, Frederick, MD 21702, USA.
⁴ University of Pittsburgh Department of Medicine, Pittsburgh, PA 15261, USA.
⁵ Children's Hospital of Philadelphia and Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA.
⁶ St. Anna Children's Cancer Research Institute, Vienna, Austria; Christian Doppler Laboratory for Next Generation CAR T Cells, Vienna, Austria.
⁷ Department of Molecular Oncology, British Columbia Cancer Research Centre, Vancouver, BC V5Z 1L3, Canada.
⁸ Program in Molecular Medicine, Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada.
⁹ BD Biosciences, San Jose, CA 95131, USA.
¹⁰ Institute of Neuropathology, University Hospital Münster, Münster, Germany.
¹¹ Center for Personal Dynamic Regulomes, Stanford University, Stanford, CA 94305, USA.
¹² Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA.
¹³ Department of Comparative Medicine, Animal Histology Services, Stanford University School of Medicine, Stanford, CA 94305, USA.
¹⁴ Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.
¹⁵ Center for Personal Dynamic Regulomes, Stanford University, Stanford, CA 94305, USA; Parker Institute for Cancer Immunotherapy, San Francisco, CA 941209, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA 94305, USA.
¹⁶ St. Anna Children's Cancer Research Institute, Vienna, Austria.
¹⁷ Program in Molecular Medicine, Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada; Departments of Biochemistry and Immunology, University of Toronto, Toronto, ON M5S 1A8, Canada.
¹⁸ Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Lorry Lokey Building, Suite G3141, MC: 5456, 265 Campus Drive, Stanford, CA 94305, USA; Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA; Parker Institute for Cancer Immunotherapy, San Francisco, CA 941209, USA; Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA. Electronic address: [email protected].

Abstract

Pediatric cancers often mimic fetal tissues and express proteins normally silenced postnatally that could serve as immune targets. We developed T cells expressing chimeric antigen receptors (CARs) targeting glypican-2 (GPC2), a fetal antigen expressed on neuroblastoma (NB) and several other solid tumors. CARs engineered using standard designs control NBs with transgenic GPC2 overexpression, but not those expressing clinically relevant GPC2 site density (∼5,000 molecules/cell, range 1-6 × 10³). Iterative engineering of transmembrane (TM) and co-stimulatory domains plus overexpression of c-Jun lowered the GPC2-CAR antigen density threshold, enabling potent and durable eradication of NBs expressing clinically relevant GPC2 antigen density, without toxicity. These studies highlight the critical interplay between CAR design and antigen density threshold, demonstrate potent efficacy and safety of a lead GPC2-CAR candidate suitable for clinical testing, and credential oncofetal antigens as a promising class of targets for CAR T cell therapy of solid tumors.

Keywords: CAR T cell; GPC2; antigen density; chimeric antigen receptor; glypican-2; immunotherapy; neuroblastoma; oncofetal antigen.

Publication types

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

MeSH terms

Animals
Cell Line, Tumor
Glypicans / immunology*
Glypicans / metabolism
Humans
Immunotherapy / methods
Immunotherapy, Adoptive*
Neuroblastoma / drug therapy*
Neuroblastoma / pathology
Receptors, Antigen, T-Cell / immunology
Receptors, Antigen, T-Cell / metabolism*
Receptors, Chimeric Antigen / immunology
T-Lymphocytes / drug effects
T-Lymphocytes / immunology
Xenograft Model Antitumor Assays / methods

Substances

Glypicans
Receptors, Antigen, T-Cell
Receptors, Chimeric Antigen

Abstract

Publication types

MeSH terms

Substances

Grants and funding