Druso Joseph E, MacPherson Maximilian B, Chia Shi B, Elko Evan, Aboushousha Reem, Seward David J, Abdelhamid Hend, Erickson Cuixia, Corteselli Elizabeth, Tarte Megan, Peng Zhihua, Bernier Daniel, Zito Ester, Shoulders Matthew D, Thannickal Victor J, Huang Steven, van der Vliet Albert, Anathy Vikas, Janssen-Heininger Yvonne M W
Departments of Pathology and Laboratory Medicine, College of Medicine, University of Vermont, Burlington, Vermont.
Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy.
Am J Respir Cell Mol Biol. 2024 Nov;71(5):589-602. doi: 10.1165/rcmb.2023-0379OC.
Changes in the oxidative (redox) environment accompany idiopathic pulmonary fibrosis (IPF). S-glutathionylation of reactive protein cysteines is a post-translational event that transduces oxidant signals into biological responses. We recently demonstrated that increases in S-glutathionylation promote pulmonary fibrosis, which was mitigated by the deglutathionylating enzyme glutaredoxin (GLRX). However, the protein targets of S-glutathionylation that promote fibrogenesis remain unknown. In the present study we addressed whether the extracellular matrix is a target for S-glutathionylation. We discovered increases in COL1A1 (collagen 1A1) S-glutathionylation (COL1A1-SSG) in lung tissues from subjects with IPF compared with control subjects in association with increases in ERO1A (endoplasmic reticulum [ER] oxidoreductin 1) and enhanced oxidation of ER-localized PRDX4 (peroxiredoxin 4), reflecting an increased oxidative environment of the ER. Human lung fibroblasts exposed to TGFB1 (transforming growth factor-β1) show increased secretion of COL1A1-SSG. Pharmacologic inhibition of ERO1A diminished the oxidation of PRDX4, attenuated COL1A1-SSG and total COL1A1 concentrations, and dampened fibroblast activation. Absence of enhanced COL1A1-SSG and overall COL1A1 secretion and promoted the activation of mechanosensing pathways. Remarkably, COL1A1-SSG resulted in marked resistance to collagenase degradation. Compared with COL1, lung fibroblasts plated on COL1-SSG proliferated more rapidly and increased the expression of genes encoding extracellular matrix crosslinking enzymes and genes linked to mechanosensing pathways. Overall, these findings suggest that glutathione-dependent oxidation of COL1A1 occurs in settings of IPF in association with enhanced ER oxidative stress and may promote fibrotic remodeling because of increased resistance to collagenase-mediated degradation and fibroblast activation.
氧化（氧化还原）环境的变化伴随着特发性肺纤维化（IPF）。反应性蛋白半胱氨酸的S-谷胱甘肽化是一种翻译后事件，可将氧化信号转化为生物学反应。我们最近证明，S-谷胱甘肽化的增加会促进肺纤维化，而谷胱甘肽去乙酰化酶谷氧还蛋白（GLRX）可减轻这种纤维化。然而，促进纤维化形成的S-谷胱甘肽化的蛋白质靶点仍然未知。在本研究中，我们探讨了细胞外基质是否是S-谷胱甘肽化的靶点。我们发现，与对照组相比，IPF患者肺组织中COL1A1（胶原蛋白1A1）的S-谷胱甘肽化（COL1A1-SSG）增加，同时ERO1A（内质网[ER]氧化还原蛋白1）增加，ER定位的PRDX4（过氧化物酶4）氧化增强，这反映了ER氧化环境的增加。暴露于TGFB1（转化生长因子-β1）的人肺成纤维细胞显示COL1A1-SSG的分泌增加。对ERO1A的药理学抑制减少了PRDX4的氧化，减弱了COL1A1-SSG和总COL1A1浓度，并抑制了成纤维细胞的激活。缺乏增强的COL1A1-SSG和总体COL1A1分泌，并促进了机械传感途径的激活。值得注意的是，COL1A1-SSG对胶原酶降解具有显著抗性。与COL1相比，接种在COL1-SSG上的肺成纤维细胞增殖更快，并增加了编码细胞外基质交联酶的基因和与机械传感途径相关的基因的表达。总体而言，这些发现表明，COL1A1的谷胱甘肽依赖性氧化发生在IPF患者中，与ER氧化应激增强有关，并且由于对胶原酶介导的降解和成纤维细胞激活的抗性增加，可能促进纤维化重塑。
