Mini-TCRs: Truncated T cell receptors to generate T cells from induced pluripotent stem cells

Shin-Ichiro Takayanagi; Bo Wang; Saki Hasegawa; Satoshi Nishikawa; Ken Fukumoto; Kohei Nakano; Sayaka Chuganji; Yuya Kato; Sanae Kamibayashi; Atsutaka Minagawa; Atsushi Kunisato; Hajime Nozawa; Shin Kaneko

doi:10.1016/j.omtm.2023.101109

Mini-TCRs: Truncated T cell receptors to generate T cells from induced pluripotent stem cells

Mol Ther Methods Clin Dev. 2023 Sep 16:31:101109. doi: 10.1016/j.omtm.2023.101109. eCollection 2023 Dec 14.

Authors

Shin-Ichiro Takayanagi^{1

2}, Bo Wang^{2

3}, Saki Hasegawa^{1

2}, Satoshi Nishikawa⁴, Ken Fukumoto^{1

2}, Kohei Nakano³, Sayaka Chuganji^{1

2}, Yuya Kato¹, Sanae Kamibayashi², Atsutaka Minagawa², Atsushi Kunisato¹, Hajime Nozawa¹, Shin Kaneko^{2

3}

Affiliations

¹ Kirin Central Research Institute, Kirin Holdings Company, Ltd., 26-1, Muraoka-Higashi 2, Fujisawa-shi, Kanagawa 251-8555, Japan.
² Shin Kaneko Laboratory, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
³ Shinobi Therapeutics, Inc., 46-29 Yoshida-Shimo-Adachi-cho, Sakyo-ku, Kyoto 606-8501, Japan.
⁴ R&D Division, Kyowa Kirin Co. Ltd, 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan.

Abstract

Allogeneic T cell platforms utilizing induced pluripotent stem cell (iPSC) technology exhibit significant promise for the facilitation of adoptive immunotherapies. While mature T cell receptor (TCR) signaling plays a crucial role in generating T cells from iPSCs, the introduction of exogenous mature TCR genes carries a potential risk of causing graft-versus-host disease (GvHD). In this study, we present the development of truncated TCRα and TCRβ chains, termed mini-TCRs, which lack variable domains responsible for recognizing human leukocyte antigen (HLA)-peptide complexes. We successfully induced cytotoxic T lymphocytes (CTLs) from iPSCs by employing mini-TCRs. Combinations of TCRα and TCRβ fragments were screened from mini-TCR libraries based on the surface localization of CD3 proteins and their ability to transduce T cell signaling. Consequently, mini-TCR-expressing iPSCs underwent physiological T cell development, progressing from the CD4 and CD8 double-positive stage to the CD8 single-positive stage. The resulting iPSC-derived CTLs exhibited comparable cytokine production and cytotoxicity in comparison to that of full-length TCR-expressing T lymphocytes when chimeric antigen receptors (CARs) were expressed. These findings demonstrate the potential of mini-TCR-carrying iPSCs as a versatile platform for CAR T cell therapy, offering a promising avenue for advancing adoptive immunotherapies.

Keywords: T-cell receptor; antigen recognition; chimeric antigen receptor T cell; cytotoxic T lymphocyte; graft-versus-host disease; induced pluripotent stem cell; mini-TCR.