Nitschkowski Dörte, Vierbuchen Tim, Heine Holger, Behrends Jochen, Reiling Norbert, Reck Martin, Rabe Klaus F, Kugler Christian, Ammerpohl Ole, Drömann Daniel, Muley Thomas, Kriegsmann Mark, Stathopoulos Georgious T, Arendt Kristina A M, Goldmann Torsten, Marwitz Sebastian
Histology, Research Center Borstel - Leibniz Lung Center, Borstel, Germany.
Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL), Borstel, Lübeck and Großhansdorf, Germany.
Br J Cancer. 2025 May 3. doi: 10.1038/s41416-025-02970-1.
We investigated the impact of SMAD2 linker phosphorylation (pSMAD2L) on overall and disease-free survival, signal transduction, as well as cancer-related processes in non-small cell lung cancer (NSCLC).
We generated A549 cells constitutively lacking pSMAD2L (A549L) to gain mechanistic insights and stimulated NSCLC cell lines with inhibitors against cell cycle-associated kinases or TGFβ1. In addition, we analysed SMAD2 and pSMAD2L in alveolar epithelial cells type 2 from tumour-free lung tissue as well as in benign and malignant T cells by Western blotting. Furthermore, pSMAD2L-positive tumours and immune cells were analysed in an NSCLC patient cohort (n = 316) using multiplex immunofluorescence.
In NSCLC cell lines and benign T cells, pSMAD2L was expressed in a mitosis-dependent manner. Loss of pSMAD2L (A549L) had an anti-proliferative effect, slowed migration, and increased alternatively spliced short SMAD2 (SMAD2). The gene signature in A549L was associated with developmental and morphogenetic processes and redirected canonical TGFβ1-dependent signalling. By contrast, SMAD2 was absent in benign T cells but present in malignant T lymphoblasts. NSCLC patients with low pSMAD2L tumour cell density had a poorer prognosis, whereas low pSMAD2L immune cell density favoured overall and disease-free survival.
pSMAD2L antagonises anti-proliferative canonical TGFβ-signalling in NSCLC and redirects TGFβ1-dependent gene expression, whereas loss of pSMAD2L enhances SMAD2 and affects pluripotency-associated proteins in vitro. In NSCLC patients, pSMAD2L cell density influences disease-free and overall survival in a spatially distinct manner.
我们研究了SMAD2连接区磷酸化（pSMAD2L）对非小细胞肺癌（NSCLC）总生存期和无病生存期、信号转导以及癌症相关过程的影响。
我们构建了组成性缺乏pSMAD2L的A549细胞（A549L），以深入了解其机制，并使用细胞周期相关激酶抑制剂或TGFβ1刺激NSCLC细胞系。此外，我们通过蛋白质免疫印迹分析了来自无肿瘤肺组织的2型肺泡上皮细胞以及良性和恶性T细胞中的SMAD2和pSMAD2L。此外，使用多重免疫荧光分析了一个NSCLC患者队列（n = 316）中的pSMAD2L阳性肿瘤和免疫细胞。
在NSCLC细胞系和良性T细胞中，pSMAD2L以有丝分裂依赖性方式表达。pSMAD2L缺失（A549L）具有抗增殖作用，减缓迁移，并增加可变剪接的短SMAD2（SMAD2）。A549L中的基因特征与发育和形态发生过程相关，并重新定向了经典的TGFβ1依赖性信号传导。相比之下，良性T细胞中不存在SMAD2，但在恶性T淋巴母细胞中存在。pSMAD2L肿瘤细胞密度低的NSCLC患者预后较差，而pSMAD2L免疫细胞密度低则有利于总生存期和无病生存期。
pSMAD2L拮抗NSCLC中抗增殖的经典TGFβ信号传导并重新定向TGFβ1依赖性基因表达，而pSMAD2L的缺失在体外增强SMAD2并影响多能性相关蛋白。在NSCLC患者中，pSMAD2L细胞密度以空间上不同的方式影响无病生存期和总生存期。
