PD-L1 transmembrane domain substitution overcomes CAR-T exhaustion driven by tonic signaling.
5 affiliations
- 1ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai 200031, China.
- 2Shanghai Key Laboratory of Gene Editing and Cell Therapy for Rare Diseases, Fudan University, Shanghai 200031, China.
- 3School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.
- 4Institute of Biomedicine Sciences & Shanghai Stomatological Hospital, Fudan University, Shanghai, China.
- 5Changping Laboratory, Beijing 102206, China.
Abstract
Chimeric antigen receptor (CAR)-T cell therapy has achieved limited success against solid tumors, in part because excessive tonic signaling drives premature T cell exhaustion, as exemplified by CSPG4 and GD2 CARs. Here, we demonstrate that replacing the dimeric CD28 transmembrane (TM) domain with the monomeric PD-L1 TM domain substantially mitigates tonic signaling. The PD-L1 TM displays a smaller tilt angle and lower hydrophobic thickness, enabling a better membrane fit that minimizes spontaneous CAR clustering. This structural feature reduces tonic signaling and exhaustion and confers superior proliferation and memory preservation, resulting in enhanced and sustained tumor killing in vitro and in vivo. Mechanistically, these benefits were associated with attenuated phosphorylation across the CD3ζ-ZAP70-PLC-γ1/MAPK/AKT/NF-κB axis, downregulation of exhaustion-associated genes, and upregulation of stemness- and memory-related programs. Collectively, our findings establish that the PD-L1 TM swap is a modular and broadly applicable strategy to suppress CAR tonic signaling and promote durable, memory-biased CAR-T responses against solid tumors.
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BibTeX
@article{zhang2026pdl1,
title = {{PD-L1 transmembrane domain substitution overcomes CAR-T exhaustion driven by tonic signaling.}},
author = {Liang Zhang and Xiang Xu and Haiwen Yu and Shizhen Qiu and Zhitong Wang and Yinying Chu and Guangfei Li and Yi Fang and Peijie He and Haitao Wu and Lei Cheng and Hongxiu Yu and Haopeng Wang and Jian Chen},
journal = {Molecular therapy. Oncology},
year = {2026},
month = oct,
doi = {10.1016/j.omton.2026.201364},
url = {https://europepmc.org/article/MED/42997611},
note = {CC BY},
}
RIS
TY - JOUR
TI - PD-L1 transmembrane domain substitution overcomes CAR-T exhaustion driven by tonic signaling.
AU - Liang Zhang
AU - Xiang Xu
AU - Haiwen Yu
AU - Shizhen Qiu
AU - Zhitong Wang
AU - Yinying Chu
AU - Guangfei Li
AU - Yi Fang
AU - Peijie He
AU - Haitao Wu
AU - Lei Cheng
AU - Hongxiu Yu
AU - Haopeng Wang
AU - Jian Chen
PY - 2026
DA - 2026/10/14
JO - Molecular therapy. Oncology
DO - 10.1016/j.omton.2026.201364
UR - https://europepmc.org/article/MED/42997611
AB - Chimeric antigen receptor (CAR)-T cell therapy has achieved limited success against solid tumors, in part because excessive tonic signaling drives premature T cell exhaustion, as exemplified by CSPG4 and GD2 CARs. Here, we demonstrate that replacing the dimeric CD28 transmembrane (TM) domain with the monomeric PD-L1 TM domain substantially mitigates tonic signaling. The PD-L1 TM displays a smaller tilt angle and lower hydrophobic thickness, enabling a better membrane fit that minimizes spontaneous CAR clustering. This structural feature reduces tonic signaling and exhaustion and confers superior proliferation and memory preservation, resulting in enhanced and sustained tumor killing in vitro and in vivo. Mechanistically, these benefits were associated with attenuated phosphorylation across the CD3ζ-ZAP70-PLC-γ1/MAPK/AKT/NF-κB axis, downregulation of exhaustion-associated genes, and upregulation of stemness- and memory-related programs. Collectively, our findings establish that the PD-L1 TM swap is a modular and broadly applicable strategy to suppress CAR tonic signaling and promote durable, memory-biased CAR-T responses against solid tumors.
SN - 2950-3299
ER -