Ligand-based, piggyBac-engineered CAR-T cells targeting EGFR are safe and effective against non-small cell lung cancers

Thanyavi Chinsuwan; Koichi Hirabayashi; Shuji Mishima; Aiko Hasegawa; Miyuki Tanaka; Hidemi Mochizuki; Akihito Shimoi; Takashi Murakami; Shigeki Yagyu; Kimihiro Shimizu; Yozo Nakazawa

doi:10.1016/j.omto.2023.100728

Ligand-based, piggyBac-engineered CAR-T cells targeting EGFR are safe and effective against non-small cell lung cancers

Mol Ther Oncolytics. 2023 Sep 16:31:100728. doi: 10.1016/j.omto.2023.100728. eCollection 2023 Dec 19.

Authors

Thanyavi Chinsuwan^{1

2}, Koichi Hirabayashi¹, Shuji Mishima³, Aiko Hasegawa¹, Miyuki Tanaka^{1

4}, Hidemi Mochizuki^{4

5}, Akihito Shimoi^{4

5}, Takashi Murakami⁶, Shigeki Yagyu^{1

4}, Kimihiro Shimizu³, Yozo Nakazawa^{1

4

7}

Affiliations

¹ Department of Pediatrics, Shinshu University School of Medicine, Matsumoto, Nagano, Japan.
² Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
³ Division of General Thoracic Surgery, Department of Surgery, Shinshu University School of Medicine, Matsumoto, Nagano, Japan.
⁴ Center for Advanced Research of Gene and Cell Therapy, Shinshu University School of Medicine, Matsumoto, Nagano, Japan.
⁵ Ina Research Inc., Ina, Nagano, Japan.
⁶ Department of Microbiology, Faculty of Medicine, Saitama Medical University, Iruma, Saitama, Japan.
⁷ Institute for Biomedical Sciences, Interdisciplinary Cluster for Cutting Edge Research, Shinshu University, Matsumoto, Nagano, Japan.

Abstract

Epidermal growth factor receptor (EGFR) is overexpressed in various cancers, including non-small cell lung cancer (NSCLC), and in some somatic cells at a limited level, rendering it an attractive antitumor target. In this study, we engineered chimeric antigen receptor (CAR)-T cells using the piggyBac transposon system, autologous artificial antigen-presenting cells, and natural ligands of EGFR. We showed that this approach yielded CAR-T cells with favorable phenotypes and CAR positivity. They exhibited potent antitumor activity against NSCLC both in vitro and in vivo. When administered to tumor-bearing mice and non-tumor-bearing cynomolgus macaques, they did not elicit toxicity despite their cross-reactivity to both murine and simian EGFRs. In total we tested three ligands and found that the CAR candidate with the highest affinity consistently displayed greater potency without adverse events. Taken together, our results demonstrate the feasibility and safety of targeting EGFR-expressing NSCLCs using ligand-based, piggyBac-engineered CAR-T cells. Our data also show that lowering the affinity of CAR molecules is not always beneficial.

Keywords: CAR-T; EGFR; NSCLC; cancer immunotherapy; non-human primate; piggyBac.