Scholarlink

<p>Acute limb ischemia (ALI) is characterized by&#xa0;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 (<i>p</i> &lt; 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 <i>proof-of-concept</i> 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.</p> Graphical abstract <p>Arterial thrombi in acute limb ischemia (ALI) exhibit a robust proteomic structure that may enhance thrombolysis resistance. Proteomic analysis revealed that ALI thrombi are highly enriched with fibrinolysis regulators, thromboinflammatory complement proteins, and platelet activators. Thrombus samples were collected post-revascularization from ALI patients, whereas in vitro control clots were generated from healthy donor blood via tissue factor‒induced coagulation. Clots were subjected to chemical and enzymatic digestion, followed by protein identification and quantification via liquid chromatography‒mass spectrometry (LC–MS). A comprehensive literature review categorized the most abundant proteins based on their roles in fibrinolysis, red blood cell degradation, complement activation, and platelet activation. ALI thrombi are highly abundant with antifibrinolytic proteins (vitronectin), thrombogenic complement proteins, platelet activators, and heme/hemoglobin scavengers. These findings reveal a fibrinolysis-resistant, prothrombotic environment, providing new insights into the mechanisms that drive clot durability and potential therapeutic targets to improve ALI management.</p> <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Proteomics of arterial thrombi in acute limb ischemia

  • 期刊论文

  • 发表日期：

    2025年6月22日

查看全文
  • Robert D. Stegman,
  • Lauren R. Schmitt,
  • Kirk C. Hansen,
  • Adrianna Kayden,
  • Ahana Ghosh,
  • Ara Metjian,
  • Max V. Wohlauer

摘要

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.

Graphical abstract

Arterial thrombi in acute limb ischemia (ALI) exhibit a robust proteomic structure that may enhance thrombolysis resistance. Proteomic analysis revealed that ALI thrombi are highly enriched with fibrinolysis regulators, thromboinflammatory complement proteins, and platelet activators. Thrombus samples were collected post-revascularization from ALI patients, whereas in vitro control clots were generated from healthy donor blood via tissue factor‒induced coagulation. Clots were subjected to chemical and enzymatic digestion, followed by protein identification and quantification via liquid chromatography‒mass spectrometry (LC–MS). A comprehensive literature review categorized the most abundant proteins based on their roles in fibrinolysis, red blood cell degradation, complement activation, and platelet activation. ALI thrombi are highly abundant with antifibrinolytic proteins (vitronectin), thrombogenic complement proteins, platelet activators, and heme/hemoglobin scavengers. These findings reveal a fibrinolysis-resistant, prothrombotic environment, providing new insights into the mechanisms that drive clot durability and potential therapeutic targets to improve ALI management.