Site-specific therapies to prevent intimal hyperplasia

项目总结： 在这个方案中，我们假设抗体介导的靶向是抗血栓/抗炎效应。 血管壁损伤部位的分子可以抑制凝血酶和嘌呤能依赖的通路，并增强 消解炎症反应。在这一过程中，功能完整的内皮细胞的重建将是 加速再狭窄，抑制再狭窄。具体来说，我们打算： (1)确定抗体导向的融合蛋白消除凝血酶依赖性炎症的能力 有助于受损的血管壁愈合的途径。在这些调查的第一阶段，我们计划 基于ScFvanti-Libs和任一血栓调节蛋白的重组抗血栓融合蛋白的研究 (TM)或直接的Xa抑制剂，Tick抗凝肽(TAP)，选择性地靶向激活的血小板并限制 体外和体内促凝血剂反应。这些研究的第二阶段将确定药物动力学特征。 这些结构，以及全身抗凝作用。在最后阶段，我们将评估在多大程度上 表面靶向TM和/或TAP通过直接注射减少ApoE/-小鼠主动脉成形术模型的内膜增生 对中膜SMC的影响或改变骨髓来源的祖细胞向SMC的招募和分化 受伤地点。 (2)确定靶向血管壁损伤部位的CD39/CD73对嘌呤能神经的限制作用。 导致血栓形成和再狭窄的介导性炎症途径。实验将寻求 检测基于单链抗体、CD39和/或CD39的重组抗血栓融合蛋白的能力 CD73选择性靶向活化的血小板并在体外介导嘌呤能通路的阻断。这些结果将是 与体内血栓形成、全身抗凝作用的存在以及血栓形成的特异性相关 血栓靶向。随后的调查将单独或在#年确定这一战略的有效性。 与TM/TAP融合蛋白结合可在体内限制血栓生长和内膜增生。这件事的影响 骨髓来源的祖细胞的动员、招募和分化的策略将是 确定的，由实验结果决定的。 (3)描述特定部位的生物活性脂类的传递促进分解的程度 损伤引起的炎性事件有助于限制内膜的增生。这些研究的第一阶段将寻求 目的：确定重组蛋白聚合物胶束在体内靶向血管壁损伤部位的能力。在……里面 在这一目标的第二阶段，实验将确定系统性与地方性交付的相对有效性 18R-HEPE、RvE1和RvD1在抑制体内内膜增生和促进内皮再生中的作用 这些介体在改变骨髓来源的祖细胞的募集和分化中的作用。 最后，我们将评估当两个促血栓形成过程都是 废除并促进加速炎症性事件解决的途径。

PROJECT SUMMARY: In this proposal, we hypothesize that antibody mediated targeting of anti-thrombotic/anti-inflammatory effector molecules to sites of vessel wall injury can inhibit thrombin and purinergic dependant pathways and enhance the resolution of the inflammatory response. In the process, the reconstitution of a functionally intact endothelium will be accelerated and restenosis inhibited. Specifically, we intend to: (1) Define the capacity of antibody-directed fusion proteins to abrogate thrombin-dependant inflammatory pathways that contribute to impaired vascular wall healing. In the first phase of these investigations, we plan to define the capacity of recombinant anti-thrombotic fusion proteins based upon scFvanti-LIBS and either thrombomodulin (TM) or the direct Xa inhibitor, tick anticoagulant peptide (TAP), to selectively target activated platelets and limit procoagulant responses in vitro and in vivo. The second phase of these studies will define the pharmacokinetic profile of these constructs, as well as systemic anticoagulant effects. In the final phase, we will evaluate the extent to which surface targeted TM and/or TAP reduce intimal hyperplasia in an ApoE-/- mouse model of aortic angioplasty by a direct effect on medial SMCs or by altering the recruitment and differentiation of bone marrow derived progenitor cells to the site of injury. (2) Determine the effectiveness of CD39/CD73 targeted to sites of vessel wall injury in limiting purinergic mediated inflammatory pathways that contribute to thrombus formation and restenosis. Experiments will seek to define the ability of recombinant anti-thrombotic fusion proteins based upon scFvanti-LIBS and either CD39 and/or CD73 to selectively target activated platelets and mediate purinergic pathway blockade in vitro. These results will be correlated with thrombus formation in vivo, the presence of systemic anticoagulant effects, and the specificity of thrombus targeting. Subsequent investigations will determine the effectiveness of this strategy either alone or in combination with TM/TAP fusion proteins to limit thrombus growth and intimal hyperplasia in vivo. The impact of this strategy on the mobilization, recruitment, and differentiation of bone marrow derived progenitor cells will be determined, as dictated by experimental findings. (3) Characterize the extent to which site-specific delivery of bioactive lipids that promote the resolution of injury induced inflammatory events serve to limit intimal hyperplasia. The first phase of these studies will seek to define the capacity of recombinant protein polymer micelles to be targeted to sites of vessel wall injury in vivo. In the second phase of this aim, experiments will determine the relative effectiveness of systemic versus local delivery of 18R-HEPE, RvE1, and RvD1 in limiting intimal hyperplasia and accelerating endothelial regeneration in vivo and the role of these mediators, if any, on altering the recruitment and differentiation of bone marrow derived progenitor cells. Finally, we will assess whether vascular wall healing can be further optimized when both pro-thrombotic processes are abrogated and pathways that accelerate the resolution of inflammatory events are promoted.