Maxwell Colleen B, Stylianou Panayiota, Marshall Hilary, Hall Alfie J, Quinn Paulene A, Ng Leong L, Jones Donald Jl, Bradding Peter, Roach Katy M
Department of Cardiovascular Sciences and NIHR Leicester Cardiovascular BRC, Glenfield Hospital, University of Leicester, Leicester, UK; Leicester van Geest MultiOMICS Facility, Hodgkin Building, University of Leicester, Leicester, UK.
Department of Respiratory Sciences, Leicester Respiratory NIHR BRC, Glenfield Hospital, University of Leicester, Leicester, UK.
Eur J Pharmacol. 2025 Jun 15;997:177461. doi: 10.1016/j.ejphar.2025.177461. Epub 2025 Mar 4.
Novel treatments for idiopathic pulmonary fibrosis (IPF) are needed urgently. A better understanding of the molecular pathways activated by TGFβ1 in human lung tissue may facilitate the development of more effective anti-fibrotic medications. This study utilized proteomic analysis to test the hypothesis that TGFβ1 induces pro-fibrotic effects on human lung parenchyma proteome, and to evaluate the viability of this model for testing novel therapeutic targets.
Non-fibrotic human lung parenchymal tissue from 11 patients was cultured for 7 days in serum-free (SF) media supplemented with TGFβ1 (10 ng/mL) or vehicle control, and the putative antifibrotic K3.1 ion channel blocker senicapoc or vehicle control. The tissue was homogenised, digested for bottom-up proteomics, and analysed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Principal component analysis, differential expression analysis, pathway analysis, and drug repurposing analysis were performed.
TGFβ1 stimulation for 7 days induced a strong fibrotic protein response relevant to IPF pathology. A total of 2391 proteins were quantified, 306 upregulated and 285 downregulated (FDR-adjusted p-value<0.05). Of these, 118 were upregulated and 28 downregulated at log(FC) > 0.58. These changes were attenuated by senicapoc (100 nM). Drug repurposing analysis identified 265 drugs predicted to inhibit the effects of TGFβ1 in this model. These included clotrimazole, a K3.1 blocker, and nintedanib, a drug licenced for the treatment of IPF, providing validation of this approach.
A pro-fibrotic proteome is induced in human lung parenchyma exposed to TGFβ1, sensitive to pharmacological intervention. This approach has the potential to enhance therapeutic drug screening for IPF treatment.
特发性肺纤维化（IPF）急需新的治疗方法。深入了解转化生长因子β1（TGFβ1）在人肺组织中激活的分子途径，可能有助于开发更有效的抗纤维化药物。本研究利用蛋白质组学分析来检验TGFβ1对人肺实质蛋白质组产生促纤维化作用的假说，并评估该模型用于测试新型治疗靶点的可行性。
将11例患者的非纤维化人肺实质组织在补充有TGFβ1（10 ng/mL）或溶剂对照的无血清（SF）培养基中培养7天，同时加入假定的抗纤维化K3.1离子通道阻滞剂司尼卡波或溶剂对照。将组织匀浆，进行自下而上的蛋白质组学消化，并使用液相色谱-串联质谱（LC-MS/MS）进行分析。进行主成分分析、差异表达分析、通路分析和药物再利用分析。
TGFβ1刺激7天诱导了与IPF病理相关的强烈纤维化蛋白反应。共定量了2391种蛋白质，其中306种上调，285种下调（FDR校正P值<0.05）。其中，118种在log(FC)>0.58时上调，28种下调。这些变化被司尼卡波（100 nM）减弱。药物再利用分析确定了265种预计在该模型中抑制TGFβ1作用的药物。其中包括K3.1阻滞剂克霉唑和已获许可用于治疗IPF的药物尼达尼布，验证了该方法的有效性。
暴露于TGFβ1的人肺实质中诱导出促纤维化蛋白质组，对药物干预敏感。这种方法有可能加强IPF治疗的治疗药物筛选。
