Myeloid TLR4 epigenetic regulation and signaling in accelerating venous thrombus resolution

项目摘要 本提案描述了血管生物学研究职业发展的5年培训计划， 重点是血栓形成候选人安德烈·奥比博士的长期目标是成为一名独立的 资助的外科医生-科学家，促进对免疫血栓形成的理解，减少深部疾病的发病率。 静脉血栓形成（DVT）。奥比博士有一个强大的基础与先进的本科教育， 微生物学和免疫学，在T32博士后期间进行显微外科静脉血栓形成模型培训 奖学金，并完成血管外科临床奖学金。一个有才华和经验丰富的导师 团队提供了详细的技能和科学知识获取计划，并提供了纵向专业知识。 发展，以确保她在开发一个独立的研究计划，在先天的交叉成功 免疫和血栓形成。 博士欧比的直接目标是利用K 08保护的时间和指导，以发展先进的专业知识 免疫学实验技术，巩固了她在调查表观遗传修饰方面的科学技能， 并提高她在骨髓细胞信号传导途径方面的知识，这将有助于发展基于免疫的 用于治疗DVT的非抗凝治疗。在临床上，存在持续残留的静脉 DVT发作后的血栓与瓣膜功能障碍、复发性血栓形成和术后 血栓综合征在实验中，先天免疫细胞，主要是单核细胞/巨噬细胞， 在解决血栓方面的作用。在一个翻译小鼠模型中，我们证明了TLR 4是一个重要的介导因子， 血栓消退。在所附提案中，我们计划解决诊所护理的一个主要差距，即 缺乏血栓消退的非抗凝分子靶点，通过评价中心假设， 髓样TLR 4介导的血栓消退是由表观遗传变化驱动的， 骨髓来源的单核细胞对急性静脉血栓形成的反应，并决定随后的局部 细胞因子和纤维蛋白溶解反应。这将通过三个具体目标来实现：（1）检查配体， TLR 4的动力学和细胞来源以及血栓消退期间对髓样TLR 4信号传导的需求;（2） 检测染色质修饰酶MLL 1对血栓形成后TLR 4表达和分子水平的影响， 血栓消退的介质;（3）确定髓系特异性TLR 4激动作为策略的可行性 加速血栓消退并评估对静脉壁损伤的影响。

PROJECT ABSTRACT This proposal describes the 5-year training program for development of a research career in vascular biology, with focus on thrombosis. The candidate, Dr. Andrea Obi, has the long-term goals of becoming an independently funded surgeon-scientist, advancing understanding of immunothrombosis and reducing the morbidity from deep venous thrombosis (DVT). Dr. Obi has a strong foundation with advanced undergraduate education in Microbiology and Immunology, training in microsurgical venous thrombosis models during a T32 post-doctoral fellowship, and completion of a clinical fellowship in Vascular Surgery. A talented and experienced mentorship team provides a detailed plan of skill and scientific knowledge acquisition, and longitudinal professional development to ensure her success in developing an independent research program at the intersection of innate immunity and thrombosis. Dr. Obi's immediate goal is to leverage the K08 protected time and mentorship to develop expertise in advanced immunologic experimental techniques, solidify her scientific skill set in investigating epigenetic modifications, and to advance her knowledge in myeloid cellular signaling pathways, that will aid in developing immune based non-anticoagulant therapies for the treatment of DVT. Clinically, the presence of persistent residual venous thrombi following an episode of DVT is associated with valvular dysfunction, recurrent thrombosis and post thrombotic syndrome. Experimentally, innate immune cells, primarily monocytes/macrophages, play a central role in resolving the thrombus. In a translational mouse model we demonstrate that TLR4 is an essential mediator of thrombus resolution. Within the enclosed proposal we plan to address a major gap in clinic care, namely a lack of non-anticoagulant molecular targets for thrombus resolution, by evaluating the central hypothesis that that myeloid TLR4-mediated thrombus resolution is mechanistically driven by epigenetic changes that occur in bone marrow derived monocytes in response to acute venous thrombosis and determines the subsequent local cytokine and fibrinolytic response. This will be accomplished via three specific aims: (1) to examine the ligands, kinetics and cellular source of TLR4 and requirement for myeloid TLR4 signaling during thrombus resolution; (2) to examine the role of chromatic modifying enzyme MLL1 on postthrombotic TLR4 expression and molecular mediators of thrombus resolution; (3) to determine the feasibility of myeloid specific TLR4 agonism as a strategy to accelerate thrombus resolution and assess impact on vein wall injury.