Sarohi Vivek, Basak Trayambak
School of Biosciences and Bioengineering (SBB), Indian Institute of Technology (IIT)- Mandi, India; BioX Center, IIT-Mandi, Himachal Pradesh 175075, India.
School of Biosciences and Bioengineering (SBB), Indian Institute of Technology (IIT)- Mandi, India; BioX Center, IIT-Mandi, Himachal Pradesh 175075, India.
J Proteomics. 2023 Mar 30;276:104842. doi: 10.1016/j.jprot.2023.104842. Epub 2023 Feb 11.
Myocardial infarction (MI) leading to heart failure contributes to almost 85% of deaths associated with CVDs. MI results from plaque formation in the coronary artery which leads to a lack of oxygen and nutrients in the myocardium. To date, stenting is a widely used gold-standard technique to maintain the proper blood flow through coronary circulation in the myocardium. Bare metal stents (BMS) and drug-eluting stents (DES) are majorly used in implantation. However, BMS and DES both can induce neointima formation by depositing excessive collagens in the coronary arteries leading to restenosis. Identification and quantitative analysis of site-specific post-translational modifications (PTMs) of deposited COL1A1 from neointima ECM are not known. Applying our in-house workflow, we re-analyzed a previously published mass-spectrometry data set to comprehensively map site-specific prolyl-hydroxylation, lysyl hydroxylation, and O-glycosylation sites in COL1A1 from neointima ECM. Furthermore, we quantitated the occupancy level of 9 3-hydroxyproline (3-HyP) sites, 2 hydroxylysine sites, and glycosylation microheterogeneity on 6 lysine sites of COL1A1. Although the total level of COL1A1 was decreased in DES-induced neointima, the occupancy levels of 2 3-HyP sites (P, and P) and 2 HyK (K and K) sites of COL1A1 were significantly (p < 0.05) elevated in DES-induced neointima compared to BMS-induced neointima. We also found O-glycosylation to be significantly elevated on 3 lysine sites (K, K, and K and K) of COL1A1 in DES-induced neointima compared to BMS-induced neointima. Taken together, our first comprehensive PTM analysis of COL1A1 reflected significant site-specific alterations that may play a very important role in the ECM remodeling during stent-induced neointima formation in MI patients. SIGNIFICANCE: The knowledge about site-specific post-translational modifications (PTMs) of collagen 1 deposited in the neointima ECM during the post-stenting restenosis process is absent. Here for the first time, we report the altered levels of COL1A1 PTMs during metal stent and drug-eluting stent-induced neointima formation. Our study showcases a novel ECM remodeling through site-specific collagen PTMs during stent-induced restenosis.
导致心力衰竭的心肌梗死（MI）导致了近85%的心血管疾病相关死亡。心肌梗死是由冠状动脉中形成斑块所致，这会导致心肌缺氧和营养物质缺乏。迄今为止，支架植入是一种广泛应用的金标准技术，用于维持心肌中冠状动脉循环的正常血流。裸金属支架（BMS）和药物洗脱支架（DES）主要用于植入。然而，BMS和DES都可通过在冠状动脉中沉积过多胶原蛋白导致内膜增生，进而引起再狭窄。目前尚不清楚来自新生内膜细胞外基质（ECM）的沉积型COL1A1的位点特异性翻译后修饰（PTM）的鉴定和定量分析情况。应用我们内部的工作流程，我们重新分析了一个先前发表的质谱数据集，以全面绘制来自新生内膜ECM的COL1A1中位点特异性脯氨酰羟化、赖氨酰羟化和O-糖基化位点。此外，我们定量了COL1A1的9个3-羟脯氨酸（3-HyP）位点、2个羟赖氨酸位点以及6个赖氨酸位点上糖基化微异质性的占据水平。尽管DES诱导的新生内膜中COL1A1的总水平降低，但与BMS诱导的新生内膜相比，DES诱导的新生内膜中COL1A1的2个3-HyP位点（P和P）和2个HyK（K和K）位点的占据水平显著升高（p<0.05）。我们还发现，与BMS诱导的新生内膜相比，DES诱导的新生内膜中COL1A1的3个赖氨酸位点（K、K以及K和K）上的O-糖基化显著升高。综上所述，我们对COL1A1的首次全面PTM分析反映了显著的位点特异性改变，这些改变可能在MI患者支架诱导的新生内膜形成过程中的ECM重塑中发挥非常重要的作用。意义：关于支架植入后再狭窄过程中沉积在新生内膜ECM中的胶原蛋白1的位点特异性翻译后修饰（PTM）的知识尚不存在。在此，我们首次报告了金属支架和药物洗脱支架诱导新生内膜形成过程中COL1A1 PTMs水平的改变。我们的研究展示了支架诱导再狭窄过程中通过位点特异性胶原蛋白PTM进行的新型ECM重塑。
