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  2. Alpha5 nicotinic acetylcholine receptor mediated immune escape of lung adenocarcinoma via STAT3/Jab1-PD-L1 signalling

Alpha5 nicotinic acetylcholine receptor mediated immune escape of lung adenocarcinoma via STAT3/Jab1-PD-L1 signalling

  • Cell Commun Signal. 2022 Aug 15;20(1):121. doi: 10.1186/s12964-022-00934-z.
Ping Zhu 1 2 Zhengxin Jin 1 Guiyu Kang 2 Yanfei Jia 1 Duanrui Liu 3 Qian Zhang 3 Feiyang Guo 4 Ying Jia 3 Yang Jiao 3 Jingtan Li 3 Haiji Sun 4 Xiaoli Ma 5 6 7
Affiliations

Affiliations

  • 1 Research Center of Basic Medicine, Central Hospital Affiliated to Shandong First Medical University, Jinan, China.
  • 2 Department of Medical Laboratory, Weifang Medical University, Weifang, China.
  • 3 Research Center of Basic Medicine, Jinan Central Hospital, Shandong University, Jinan, China.
  • 4 College of Life Science, Shandong Normal University, Jinan, China.
  • 5 Research Center of Basic Medicine, Central Hospital Affiliated to Shandong First Medical University, Jinan, China. Maxiaoli_jn@sdu.edu.cn.
  • 6 Department of Medical Laboratory, Weifang Medical University, Weifang, China. Maxiaoli_jn@sdu.edu.cn.
  • 7 Research Center of Basic Medicine, Jinan Central Hospital, Shandong University, Jinan, China. Maxiaoli_jn@sdu.edu.cn.
Abstract

Background: Immunotherapy has proven to be an emerging treatment for non-small-cell lung Cancer in recent years. Notably, smokers show higher programmed cell death ligand-1 (PD-L1) expression and better responses to PD-1/PD-L1 inhibitors than nonsmokers. Genome-wide association studies show that the CHRNΑ5 encoding α5-nicotinic acetylcholine receptor (α5-nAChR) is especially relevant to lung Cancer and nicotine dependence. Jab1 is a key regulatory factor and promotes the stabilization of PD-L1. Our previous study reported that α5-nAChR mediates lung adenocarcinoma (LUAD) epithelial-mesenchymal transition (EMT) and metastasis via STAT3/Jab1. However, the link between α5-nAChR and PD-L1 is unclear in LUAD.

Methods: We used various bioinformatics databases to analyze the expression of related genes and their correlations. Expression and clinicopathologic significance of α5-nAChR and PD-L1 were detected by immunohistochemistry in a tissue microarray. α5-nAChR regulated LUAD cell immune escape by targeting the STAT3/Jab1-PD-L1 signalling by Western-blotting and ChIP in vitro. We used T cell coculture, flow cytometry, ELISA, CCK8 assay and crystal violet staining to detect the expression of regulatory T cell (Tregs), IFN-γ, IL-2 and the ability of T cell-mediated tumour cell killing respectively. IF assays were performed in both Cancer cells and tumour xenograft paraffin sections to analyze the protein expression. The in vivo experiments in mouse model were performed to show the α5-nAChR-mediated immune escape via PD-L1 pathway.

Results: The expression of α5-nAChR was correlated with PD-L1 expression, smoking status and lower survival of LUAD in vivo. In vitro, the expression of α5-nAChR mediated phosphorylated STAT3 (pSTAT3), Jab1 and PD-L1 expression. STAT3 bound to the Jab1 or PD-L1 promoter and mediated PD-L1 expression. Jab1 stabilized PD-L1 expression in LUAD cells. Furthermore, in primary T cell cocultured system, downregulation of α5-nAChR suppressed the function of CD4+CD25+FOXP3+ Tregs, enhanced IFN-γ secretion, and increased T cell-mediated killing of LUAD cells. In the Jurkat T cells and LUAD cells coculture assay, inhibition of α5-nAChR increased IL-2 secretion. In tumour xenograft tissues, α5-nAChR expression was related to PD-L1, Jab1, pSTAT3, CD4 and granzyme B expression (GB).

Conclusions: Our results suggest that the novel α5-nAChR/STAT3-Jab1-PD-L1 axis is involved in LUAD immune escape, which could lead to potential therapeutic strategies for Cancer Immunotherapy. Video abstract.

Keywords

Immune escape; Lung adenocarcinoma; STAT3/Jab1-PD-L1; α5-nAChR.

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