Mechanisms of vascular dysfunction in acute systemic inflammation

项目概要： 全身性炎症反应综合征，由宿主对炎症触发物的反应驱动， 外来病原体和细胞因子引起脉管系统的显著功能障碍。这种功能障碍是由于 内皮内稳态机制的丧失，表现为血管内液体丢失、异常 白细胞运输，破坏凝血和改变血管张力。尽管它的压倒性和明显的 对患者及其结局的负面影响，靶向血管通路，如血管扩张剂一氧化氮， 没有取得成功的结果。这种试验的失败，沿着普遍缺乏有效的 急性全身性炎症的治疗，除了支持性治疗外， 在乎不幸的是，急性全身性炎症中血管功能障碍的根本原因仍然完全是 未知的和许多未回答的问题。什么时空蛋白质相互作用或代谢途径 有助于或抵消功能障碍？血管病变的存在是否会导致特定的基因组或 蛋白质组特征，即内型，以及如何对它们进行建模？什么独特的内皮靶点 存在可以用来改善血管功能和恢复体内平衡的药物吗？为了回答这些问题， 今后，我们的研究计划将集中在三个综合主题上。主题一将探索 在急性炎症过程中内皮稳态失调的分子机制。等机制 作为直接的蛋白质-蛋白质相互作用， 探索使用各种技术，包括基因改造，糖酵解和氧化应激能力， 邻位连接测定。主题二将集中于利用重叠模型内皮功能障碍 在动物和人类中的程序，以确定一致的模式。这个主题将测试动物模型， 使用非侵入性血管反应性与急性病患者组合的全身性炎症 技术，如激光多普勒灌注监测，加上基因组和蛋白质组签名。在 此外，微流体装置（即，芯片上组织）的使用将在更适应性的 动物模型和非适应性人类患者群体，以测试合成人类系统是否与 数据来源于机制驱动的动物研究。第三个主题将侧重于药物发现。这 主题将使用先前主题中发现的生化机制来帮助发现内皮特异性， 有针对性的治疗使用与新化合物或肽序列偶联的细胞穿透肽将 允许细胞渗透到感兴趣的目标。此外，对慢性血管功能障碍的治疗方法的测试 将进行检查，以确定类似的机制是否可以在急性全身性血管病变中得到缓解。这些 综合主题支持这项研究计划的总体目标，即更好地了解 影响急性内皮介导的血管功能障碍的机制，总体目的是能够 识别和治疗全身炎症期间的急性血管病变患者，以改善临床结局。

PROJECT SUMMARY: The systemic inflammatory response syndrome, driven by a host’s response to inflammatory triggers such as foreign pathogens and cytokines, causes significant dysfunction in the vasculature. This dysfunction is due to the loss of homeostatic mechanisms within the endothelium and manifests as intravascular fluid loss, abnormal leukocyte trafficking, disrupted coagulation and altered vascular tone. Despite its overwhelming and obvious negative effect on patients and their outcomes, targeting vascular pathways, such the vasodilator nitric oxide, has not yielded successful results. The failure of such trials, along with the general absence of effective treatments for acute systemic inflammation, has left clinicians with no therapeutic options beyond supportive care. Unfortunately, the root cause of vascular dysfunction in acute systemic inflammation remains completely unknown and with many unanswered questions. What spatiotemporal protein interactions or metabolic pathways contribute to or counterbalance the dysfunction? Does the presence of vasculopathy lead to specific genomic or proteomic signatures, known as endotypes, and how can they be modeled? What unique endothelial targets exist that can be utilized to improve vascular function and restore homeostasis? To answer these questions and any further that will arise, our research program will focus on three integrated themes. Theme one will explore molecular mechanisms that dysregulate endothelial homeostasis during acute inflammation. Mechanisms such as direct protein-protein interactions, mitochondrial dysfunction and reactive oxygen species signaling will be explored using a variety of techniques including genetic modification, glycolytic and oxidative stress capacity and proximity ligation assays. Theme two will focus on modeling endothelial dysfunction utilizing overlapping procedures in both animals and humans to identify consistent patterns. This theme will test animal models of systemic inflammation in combination with acutely ill human patients using non-invasive vascular reactivity techniques, such as laser doppler perfusion monitoring, coupled with genomic and proteomic signatures. In addition, the use of microfluidic devices (i.e. tissue-on-a-chip) will create a bridge between the more adaptable animal models and the non-adaptable human patient populations to test if a synthetic human system will correlate with data derived from mechanism driven animal studies. The third theme will focus on drug discovery. This theme will use the biochemical mechanisms found in the prior themes to help discover endothelial-specific, targeted treatments. Use of cell-penetrating peptides coupled to novel compounds or peptide sequences will allow cell permeation to the target of interest. In addition, testing of therapies used in chronic vascular dysfunction will be examined to determine if similar mechanisms can be tempered in acute systemic vasculopathy. These integrated themes support the overarching goal of this research program, which is to better understand mechanisms that affect acute endothelial-mediated vascular dysfunction with the overall intent of being able to identify and treat patients with acute vasculopathy during systemic inflammation to improve clinical outcomes.