Stegman Robert D, Schmitt Lauren R, Hansen Kirk C, Kayden Adrianna, Ghosh Ahana, Metjian Ara, Wohlauer Max V
University of Colorado School of Medicine, 13001 E 17th Pl, Aurora, CO, 80045, USA.
Departments of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, 12801 East 17th Avenue, Aurora, CO, 80045, USA.
J Thromb Thrombolysis. 2025 Jun 22. doi: 10.1007/s11239-025-03123-0.
Acute limb ischemia (ALI) is characterized by a sudden decrease in limb perfusion due to arterial occlusion. Without urgent revascularization, patients are at risk of ischemic damage and amputation. This study uses novel proteomic techniques to investigate the molecular architecture of ALI thrombi, identifying key proteins that may influence coagulation dynamics and fibrinolysis resistance. Arterial thromboemboli (n = 12) collected after revascularization procedures were analyzed and compared to in vitro clots (n = 10) generated from healthy donor blood via tissue factor‒induced coagulation. Proteins were identified and quantified via liquid chromatography‒mass spectrometry (LC-MS/MS). A comprehensive literature review of the most abundant proteins allowed for categorization based on their functional roles in fibrinolysis, red blood cell (RBC) degradation, complement activation, and platelet activation. Compared to in vitro clots, ALI clots contained 141 proteins with significantly increased abundance (fold change) and 38 with decreased abundance (p < 0.05). These include 17 fibrinolysis regulators, 8 RBC-related proteins, 6 complement proteins, and 36 platelet regulators. The antifibrinolytic protein vitronectin (VTN) was strikingly enriched (1067-fold), suggesting a substantial role in fibrinolysis resistance and clot stability. Scavengers of heme/hemoglobin, thromboinflammatory complement proteins, and platelet activators were highly abundant. This proof-of-concept study introduces novel proteomic methods for arterial thrombus analysis and identifies key proteins involved in ALI pathology. Our findings reveal a delicate balance between antifibrinolytic and profibrinolytic proteins, offering potential therapeutic targets to enhance thrombolysis and improve ALI management.
急性肢体缺血（ALI）的特征是由于动脉闭塞导致肢体灌注突然减少。若不进行紧急血运重建，患者有发生缺血性损伤和截肢的风险。本研究采用新型蛋白质组学技术研究ALI血栓的分子结构，确定可能影响凝血动力学和纤维蛋白溶解抵抗的关键蛋白。对血运重建术后收集的动脉血栓栓子（n = 12）进行分析，并与通过组织因子诱导凝血从健康供体血液中生成的体外凝块（n = 10）进行比较。通过液相色谱 - 质谱联用（LC-MS/MS）鉴定和定量蛋白质。基于对最丰富蛋白质的全面文献综述，根据它们在纤维蛋白溶解、红细胞（RBC）降解、补体激活和血小板激活中的功能作用进行分类。与体外凝块相比，ALI凝块含有141种丰度显著增加（倍数变化）的蛋白质和38种丰度降低的蛋白质（p < 0.05）。这些包括17种纤维蛋白溶解调节因子、8种与RBC相关的蛋白质、6种补体蛋白和36种血小板调节因子。抗纤维蛋白溶解蛋白玻连蛋白（VTN）显著富集（1067倍），表明其在纤维蛋白溶解抵抗和凝块稳定性中起重要作用。血红素/血红蛋白清除剂、血栓炎症补体蛋白和血小板激活剂含量很高。这项概念验证研究引入了用于动脉血栓分析的新型蛋白质组学方法，并确定了参与ALI病理过程的关键蛋白。我们的研究结果揭示了抗纤维蛋白溶解蛋白和促纤维蛋白溶解蛋白之间的微妙平衡，为增强溶栓作用和改善ALI治疗提供了潜在的治疗靶点。
