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Single-Cell and TCR Profiling across Tissues Reveals GZMK⁺CD8⁺ T Cells as Drivers of Fibrosis in IPF

Yue, B.; Jing, S.; Li, Q.; Xie, G.; Chen, J.; Yan, T.; Zhu, G.; Huang, B.; Zhao, J.; Yang, X.; Huang, J.; Yin, P.; Yao, Q.; Yang, H.; Gao, F.; Yang, J.; Huang, M.; Chen, J.

2025-12-22 respiratory medicine
10.64898/2025.12.20.25342718 medRxiv
Show abstract

Idiopathic pulmonary fibrosis (IPF) is a fatal interstitial lung disease with limited therapies and poorly defined cross-tissue immune mechanisms. We performed single-cell RNA sequencing and TCR profiling of paired lung, lymph node, and peripheral blood samples from patients with IPF, combined with functional coculture assays and mouse model. We identified GZMK-high CD8 T cells enriched in fibrotic lungs, displaying inflammatory but low-cytotoxic features. TCR and trajectory analyses indicated that these cells originate from lymph-node CD8_HSPA1A cells and migrate to the lung. Higher GZMKCD8 T-cell levels correlated with impaired lung function and worse outcomes. Mechanistically, GZMK-overexpressing CD8 T cells promoted fibroblast-to-myofibroblast differentiation and proliferation through TGF-{beta}1 signalling, while GZMK inhibition reduced fibroblast activation and collagen deposition. Pharmacological blockade of TGF{beta}R1/ALK4 with TEW suppressed fibroblast activation in vitro and significantly attenuated pulmonary fibrosis in vivo. Together, these findings identify a lymph node-to-lung migration axis of GZMKCD8 T cells that drives fibroblast activation through TGF{beta}R1/ALK4 signalling, highlighting GZMK as a potential biomarker and therapeutic target in IPF.

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