Lorentzen Lasse G, Yeung Karin, Eldrup Nikolaj, Eiberg Jonas P, Sillesen Henrik H, Davies Michael J
Department of Biomedical Sciences, Panum Institute, University of Copenhagen, Denmark.
Department of Vascular Surgery, Heart Centre, University Hospital Copenhagen - Rigshospitalet, Copenhagen, Denmark.
Matrix Biol Plus. 2024 Jan 11;21:100141. doi: 10.1016/j.mbplus.2024.100141. eCollection 2024 Feb.
Cardiovascular disease is the leading cause of death, with atherosclerosis the major underlying cause. While often asymptomatic for decades, atherosclerotic plaque destabilization and rupture can arise suddenly and cause acute arterial occlusion or peripheral embolization resulting in myocardial infarction, stroke and lower limb ischaemia. As extracellular matrix (ECM) remodelling is associated with plaque instability, we hypothesized that the ECM composition would differ between plaques. We analyzed atherosclerotic plaques obtained from 21 patients who underwent carotid surgery following recent symptomatic carotid artery stenosis. Plaques were solubilized using a new efficient, single-step approach. Solubilized proteins were digested to peptides, and analyzed by liquid chromatography-mass spectrometry using data-independent acquisition. Identification and quantification of 4498 plaque proteins was achieved, including 354 ECM proteins, with unprecedented coverage and high reproducibility. Multidimensional scaling analysis and hierarchical clustering indicate two distinct clusters, which correlate with macroscopic plaque morphology (soft/unstable versus hard/stable), ultrasound classification (echolucent versus echogenic) and the presence of hemorrhage/ulceration. We identified 714 proteins with differential abundances between these groups. Soft/unstable plaques were enriched in proteins involved in inflammation, ECM remodelling, and protein degradation (e.g. matrix metalloproteinases, cathepsins). In contrast, hard/stable plaques contained higher levels of ECM structural proteins (e.g. collagens, versican, nidogens, biglycan, lumican, proteoglycan 4, mineralization proteins). These data indicate that a single-step proteomics method can provide unique mechanistic insights into ECM remodelling and inflammatory mechanisms within plaques that correlate with clinical parameters, and help rationalize plaque destabilization. These data also provide an approach towards identifying biomarkers for individualized risk profiling of atherosclerosis.
心血管疾病是主要的死亡原因，动脉粥样硬化是其主要潜在病因。虽然动脉粥样硬化斑块通常数十年无症状，但斑块不稳定和破裂可能突然发生，导致急性动脉闭塞或外周栓塞，进而引发心肌梗死、中风和下肢缺血。由于细胞外基质（ECM）重塑与斑块不稳定相关，我们推测不同斑块的ECM组成会有所不同。我们分析了21例近期有症状性颈动脉狭窄并接受颈动脉手术患者的动脉粥样硬化斑块。采用一种新的高效单步方法溶解斑块。将溶解的蛋白质消化成肽段，通过液相色谱 - 质谱联用仪采用数据非依赖采集模式进行分析。实现了对4498种斑块蛋白的鉴定和定量，其中包括354种ECM蛋白，具有前所未有的覆盖范围和高重现性。多维尺度分析和层次聚类表明存在两个不同的簇，这与宏观斑块形态（软/不稳定与硬/稳定）、超声分类（无回声与有回声）以及出血/溃疡的存在相关。我们鉴定出这两组之间丰度有差异的714种蛋白质。软/不稳定斑块富含参与炎症、ECM重塑和蛋白质降解的蛋白质（如基质金属蛋白酶、组织蛋白酶）。相比之下，硬/稳定斑块含有更高水平的ECM结构蛋白（如胶原蛋白、多功能蛋白聚糖、巢蛋白、双糖链蛋白聚糖、光蛋白聚糖、蛋白聚糖4、矿化蛋白）。这些数据表明，单步蛋白质组学方法可以为斑块内与临床参数相关的ECM重塑和炎症机制提供独特的机制性见解，并有助于解释斑块不稳定的原因。这些数据还提供了一种识别生物标志物以进行动脉粥样硬化个体化风险评估的方法。
