Pagano Francesca, Picchio Vittorio, Bordin Antonella, Cavarretta Elena, Nocella Cristina, Cozzolino Claudia, Floris Erica, Angelini Francesco, Sordano Alessia, Peruzzi Mariangela, Miraldi Fabio, Biondi-Zoccai Giuseppe, De Falco Elena, Carnevale Roberto, Sciarretta Sebastiano, Frati Giacomo, Chimenti Isotta
Institute of Biochemistry and Cell Biology, National Council of Research (IBBC-CNR), Monterotondo, Italy.
Department of Medical Surgical Sciences and Biotechnologies, Sapienza University, Latina, Italy.
J Pathol. 2022 Oct;258(2):136-148. doi: 10.1002/path.5985. Epub 2022 Jul 28.
Cardiac stromal cells (CSCs) are the main players in fibrosis. Dysmetabolic conditions (metabolic syndrome-MetS, and type 2 diabetes mellitus-DM2) are strong pathogenetic contributors to cardiac fibrosis. Moreover, modulation of the oxidative state (OxSt) and autophagy is a fundamental function affecting the fibrotic commitment of CSCs, that are adversely modulated in MetS/DM2. We aimed to characterize CSCs from dysmetabolic patients, and to obtain a beneficial phenotypic setback from such fibrotic commitment by modulation of OxSt and autophagy. CSCs were isolated from 38 patients, stratified as MetS, DM2, or controls. Pharmacological modulation of OxSt and autophagy was obtained by treatment with trehalose and NOX4/NOX5 inhibitors (TREiNOX). Flow-cytometry and real-time quantitative polymerase chain reaction (RT-qPCR) analyses showed significantly increased expression of myofibroblasts markers in MetS-CSCs at baseline (GATA4, ACTA2, THY1/CD90) and after starvation (COL1A1, COL3A1). MetS- and DM2-CSCs displayed a paracrine profile distinct from control cells, as evidenced by screening of 30 secreted cytokines, with a significant reduction in vascular endothelial growth factor (VEGF) and endoglin confirmed by enzyme-linked immunoassay (ELISA). DM2-CSCs showed significantly reduced support for endothelial cells in angiogenic assays, and significantly increased H O release and NOX4/5 expression levels. Autophagy impairment after starvation (reduced ATG7 and LC3-II proteins) was also detectable in DM2-CSCs. TREiNOX treatment significantly reduced ACTA2, COL1A1, COL3A1, and NOX4 expression in both DM2- and MetS-CSCs, as well as GATA4 and THY1/CD90 in DM2, all versus control cells. Moreover, TREiNOX significantly increased VEGF release by DM2-CSCs, and VEGF and endoglin release by both MetS- and DM2-CSCs, also recovering the angiogenic support to endothelial cells by DM2-CSCs. In conclusion, DM2 and MetS worsen microenvironmental conditioning by CSCs. Appropriate modulation of autophagy and OxSt in human CSCs appears to restore these features, mostly in DM2-CSCs, suggesting a novel strategy against cardiac fibrosis in dysmetabolic patients. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.
心脏基质细胞（CSCs）是纤维化的主要参与者。代谢紊乱状况（代谢综合征-MetS和2型糖尿病-DM2）是心脏纤维化的重要致病因素。此外，氧化状态（OxSt）和自噬的调节是影响CSCs纤维化进程的一项基本功能，而在MetS/DM2中它们受到不利调节。我们旨在对代谢紊乱患者的CSCs进行特征描述，并通过调节OxSt和自噬从这种纤维化进程中获得有益的表型逆转。从38例患者中分离出CSCs，分为MetS组、DM2组或对照组。通过用海藻糖和NOX4/NOX5抑制剂（TREiNOX）处理实现对OxSt和自噬的药理学调节。流式细胞术和实时定量聚合酶链反应（RT-qPCR）分析显示，在基线时（GATA4、ACTA2、THY1/CD90）以及饥饿后（COL1A1、COL3A1），MetS-CSCs中成肌纤维细胞标志物的表达显著增加。通过对30种分泌细胞因子的筛选证明，MetS-和DM2-CSCs表现出与对照细胞不同的旁分泌特征，酶联免疫吸附测定（ELISA）证实血管内皮生长因子（VEGF）和内皮糖蛋白显著减少。在血管生成试验中，DM2-CSCs对内皮细胞的支持作用显著降低，且H₂O释放和NOX4/5表达水平显著增加。在DM2-CSCs中也可检测到饥饿后自噬受损（ATG7和LC₃-II蛋白减少）。与对照细胞相比，TREiNOX处理显著降低了DM2-和MetS-CSCs中ACTA2、COL1A1、COL3A1和NOX4的表达，以及DM2中GATA4和THY1/CD90的表达。此外，TREiNOX显著增加了DM2-CSCs释放的VEGF，以及MetS-和DM2-CSCs释放的VEGF和内皮糖蛋白，还恢复了DM2-CSCs对内皮细胞的血管生成支持作用。总之，DM2和MetS会使CSCs的微环境条件恶化。对人CSCs中自噬和OxSt进行适当调节似乎可以恢复这些特征，在DM2-CSCs中最为明显，这提示了一种针对代谢紊乱患者心脏纤维化的新策略。© 2022作者。《病理学杂志》由约翰·威利父子有限公司代表大不列颠及爱尔兰病理学会出版。
