Artificial T Cell Adaptor Molecule-Transduced TCR-T Cells Demonstrated Improved Proliferation Only When Transduced in a Higher Intensity

Toshiyasu Sakai; Seitaro Terakura; Kotaro Miyao; Shingo Okuno; Yoshitaka Adachi; Koji Umemura; Jakrawadee Julamanee; Keisuke Watanabe; Hiroshi Hamana; Hiroyuki Kishi; Judith Leitner; Peter Steinberger; Tetsuya Nishida; Makoto Murata; Hitoshi Kiyoi

doi:10.1016/j.omto.2020.08.014

Artificial T Cell Adaptor Molecule-Transduced TCR-T Cells Demonstrated Improved Proliferation Only When Transduced in a Higher Intensity

Mol Ther Oncolytics. 2020 Aug 28:18:613-622. doi: 10.1016/j.omto.2020.08.014. eCollection 2020 Sep 25.

Authors

Toshiyasu Sakai¹, Seitaro Terakura¹, Kotaro Miyao¹, Shingo Okuno¹, Yoshitaka Adachi¹, Koji Umemura¹, Jakrawadee Julamanee^{1

2}, Keisuke Watanabe^{1

3}, Hiroshi Hamana⁴, Hiroyuki Kishi⁴, Judith Leitner⁵, Peter Steinberger⁵, Tetsuya Nishida¹, Makoto Murata¹, Hitoshi Kiyoi¹

Affiliations

¹ Department of Hematology and Oncology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
² Division of Clinical Hematology, Faculty of Medicine, Prince of Songkla University, Songkhla, Thailand.
³ Division of Cancer Immunology, National Cancer Center Research Institute, Tokyo, Japan.
⁴ Department of Immunology, Faculty of Medicine, Academic Assembly, University of Toyama, Toyama, Japan.
⁵ Division for Immune Receptors and T Cell Activation, Institute of Immunology, Medical University of Vienna, Vienna, Austria.

Abstract

An artificial T cell adaptor molecule (ATAM) was generated to improve persistence of T cell receptor (TCR) gene-transduced T (TCR-T) cells compared to such persistence in a preceding study. ATAMs are gene-modified CD3ζ with the intracellular domain of 4-1BB inserted in the middle of CD3ζ. NY-ESO-1 TCR-T cells transduced with an ATAM with two separated virus vectors demonstrated superior proliferation upon antigen stimulation. To further develop clinically applicable ATAM-transduced TCR-T cells, we attempted to make a single virus vector to transduce the TCR and ATAM simultaneously. Because we failed to observe improved proliferation capacity upon stimulation after one virus vector (1vv) transduction, we compared TCR-T cells transduced with 1vv and two virus vector (2vv) methods to elucidate the reason. In Jurkat reporter cells, an ATAM transduced by the 2vv method demonstrated a higher intensity than by the 1vv method, and the ATAM intensity was associated with increased nuclear factor κB (NF-κB) signals upon stimulation. In ATAM-transduced primary T cells, a transduced ATAM by the 2vv method showed higher intensity and better proliferation. ATAM-transduced TCR-T cells demonstrated improved proliferation only when the ATAM was transduced at a higher intensity. To create a simpler transduction method, we need to develop a strategy to make a higher ATAM expression to prove the efficacy of ATAM transduction in TCR-T therapy.

Keywords: CD3ζ gene modification; T cell receptor gene insertion; adaptor molecule gene-insertion intensity; artificial adaptor molecule; gene-modified T cell therapy.