Role of glycosaminoglycans (GAGs) in deep vein thrombus formation and resolution

项目总结 静脉血栓栓塞症是心血管疾病的第三大常见死亡原因 疾病。尽管在标准的抗凝治疗中增加了导管定向溶栓治疗，但复发 深静脉血栓形成(DVT)和随后的血栓形成后综合征(PTS)，其中的体征和症状 DVT的持续或恶化随着血栓的溶解，仍然会导致长期的残疾。因此，更好地理解 需要了解深静脉血栓形成和解决的基本机制。 为了形成深静脉血栓，必须发生内皮细胞(EC)损伤。最近，ECGCX，一种 糖蛋白、蛋白多糖和相关的糖胺聚糖(GAG)的膜结合网状物已经出现 作为一种多功能表层，能够调节血管扩张、炎症、增殖和 凝血途径。3个最突出的eGCX GAG，硫酸乙酰肝素(HS)，硫酸软骨素(CS)，以及 透明质酸(HA)与抗凝血酶、肝素辅因子II和非凝血酶的结合部位有关。 循环系数XI(FXI)。这些关键凝血成分的GAG结合表明 目前，GAG在血栓形成和溶解中的作用尚不清楚，因此有必要进一步 机械调查。 这项建议的目的是阐明eGCX调节DVT形成和 决议。我的假设是，受损的eGCX将通过增加血小板-EC来促进DVT的形成 相互作用，并通过释放非循环FXI激活内在凝血途径。此外， 受损的eGCX将通过增加炎性(M1)单核细胞对血栓的渗透而损害DVT的分辨率 和静脉壁。在目标1中，我将确定eGCX GAG在血小板激活和血栓形成中的作用 在野生型CD1小鼠中建立下腔静脉缩窄DVT模型。在目标2中，我将确定eGCX GAG的角色 导致血栓形成后静脉壁的改变。这项研究可能会确定相关的治疗靶点 直接影响DVT的形成和解决，为介入研究铺平道路，并支持 开发选择性、安全和有效的抗血栓药物的基本原理。这些研究将是 在俄勒冈州的主要学术医疗中心，俄勒冈健康与科学大学表演 在Monica Hinds博士的共同指导下，生物医学工程和血管外科系， 博士和Khanh Nguyen博士，医学博士PI由一个由资深科学家和医生组成的指导团队提供支持 具有血管病理生理学、血栓形成和血管外科方面的专业知识。职业发展活动 包括小鼠手术、体外和体内分析技术以及凝血和血管方面培训 病理生理学，以及沟通和领导技能的培训。本培训旨在支持 PI的职业目标是成为一名内科科学家，长期的职业目标是领导一个富有成效和 翻译血管病理生理学实验室与紧密结合的血管外科实践。

PROJECT SUMMARY Venous thromboembolism disorders represent the third most common cause of death from cardiovascular disease. Despite the addition of catheter directed thrombolysis to standard anticoagulation therapy, recurrent deep vein thrombosis (DVT) and subsequent post-thrombotic-syndrome (PTS), where the signs and symptoms of DVT persist or worsen with thrombus resolution, still cause long term disability. Thus, a better understanding of the basic mechanisms of DVT formation and resolution is needed. In order for a DVT to form, endothelial cell (EC) injury must occur. Recently, the EC glycocalyx (eGCX), a membrane bound mesh of glycoproteins, proteoglycans, and associated glycosaminoglycans (GAGs), has come into focus as a multifunctional surface layer capable of regulating vasodilation, inflammation, proliferation, and coagulation pathways. The 3 most prominent eGCX GAGs, heparan sulfate (HS), chondroitin sulfate (CS), and hyaluronan (HA), have been associated with binding sites for anti-thrombin, heparin cofactor II, and non- circulating factor XI (FXI). GAG binding of these critical coagulation components suggests an ambiguous and currently unclarified role for GAGs in thrombus formation and resolution, warranting further mechanistic investigation. The goal of this proposal is to elucidate the mechanistic role by which the eGCX regulates DVT formation and resolution. My hypothesis maintains that an injured eGCX will bolster DVT formation by increasing platelet-EC interactions and activating the intrinsic clotting pathway, via release of non-circulating FXI. Furthermore, the injured eGCX will impair DVT resolution by increasing inflammatory (M1) monocyte infiltration into the thrombus and the vein wall. In Aim 1, I will determine the role of eGCX GAGs in platelet activation and thrombus formation using a caval constriction DVT model in wildtype CD1 mice. In Aim 2, I will determine the role of eGCX GAGs in causing post-thrombotic vein wall changes. This study will potentially identify relevant therapeutic targets that directly affect DVT formation and resolution, paving the way for interventional studies and supporting the rationale for the development of selective, safe, and effective antithrombotic agents. These studies will be performed at Oregon's primary academic medical center, the Oregon Health and Science University in the Departments of Biomedical Engineering and Vascular Surgery under the co-mentorship of Dr. Monica Hinds, PhD and Dr. Khanh Nguyen, MD. The PI is supported by a mentorship team of senior scientists and physicians with expertise in vascular pathophysiology, thrombosis, and vascular surgery. Career development activities include training in mouse surgery, in vitro and in vivo analysis techniques, and coagulation and vascular pathophysiology, as well as training in communication and leadership skills. This training is designed to support the PI's career goals of becoming a physician-scientist with a long-term career goal of leading a productive and translational vascular pathophysiology laboratory with a tightly integrated vascular surgery practice.