Myeloid TLR4 epigenetic regulation and signaling in accelerating venous thrombus resolution

项目摘要 这份提案描述了血管生物学研究生涯发展的5年培训计划， 重点是血栓形成。候选人安德里亚·奥比博士的长期目标是成为一名独立的 资助外科医生和科学家，提高对免疫血栓形成的认识，从深层次降低发病率 静脉血栓(DVT)。欧比博士拥有雄厚的基础，在 微生物学和免疫学，T32博士后显微外科静脉血栓模型培训 奖学金，并完成血管外科临床奖学金。才华横溢、经验丰富的导师关系 团队提供详细的技能和科学知识获取计划，并纵向专业 开发以确保她在先天的交汇处成功地开发了一个独立的研究计划 免疫力和血栓形成。 欧比博士的直接目标是利用K08受保护的时间和指导来开发高级专业知识 免疫学实验技术，巩固了她研究表观遗传修饰的科学技能， 并提高她在髓系细胞信号通路方面的知识，这将有助于开发基于 深静脉血栓的非抗凝治疗。临床上，持续残存静脉的存在 DVT发作后的血栓与瓣膜功能障碍、复发的血栓形成和 血栓综合征。实验上，先天免疫细胞，主要是单核/巨噬细胞，在 在解决血栓中的作用。在翻译小鼠模型中，我们证明了TLR4是一个重要的介体 血栓消退的可能性。在随附的建议中，我们计划解决临床护理方面的一个主要差距，即 缺乏用于血栓溶解的非抗凝分子靶点，通过评估以下中心假设 髓系TLR4介导的血栓溶解是由发生在 骨髓来源的单核细胞对急性静脉血栓的反应并确定随后的局部 细胞因子和纤溶反应。这将通过三个具体目标来实现：(1)检查配体， 血栓溶解过程中TLR4的动力学和细胞来源以及对髓系TLR4信号的需求； 染色单体修饰酶MLL1对血栓后TLR4表达及分子水平的影响 血栓溶解的介体；(3)确定髓系特异性TLR4激动剂作为策略的可行性 加速血栓溶解，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.