Targeted Suppression of Cytokine Signaling in the Acute Phase Response

描述（由申请人提供）：炎症作为疾病的主要机制，是脓毒症（“脓毒性休克”）中血管系统塌陷和动脉粥样硬化发展的基础。两种疾病过程在60-85岁年龄组中显著会聚，其中脓毒症相关死亡率达到45%，而所有年龄组的死亡率为30%。促炎细胞因子诱导急性期蛋白反应（APPR）。APPR表现为生物标志物C-反应蛋白（CRP）的表达，以及血清淀粉样蛋白、补体蛋白、凝血因子和蛋白酶标志物水平的升高-每种都有助于心血管炎性损伤。CRP和其他与炎症过程相关的生物标志物已成为最近流行病学和临床研究的焦点。在美国，每年有100万心脏病发作，783，000中风和估计750，000败血症患者住院。高胆固醇西方饮食和其他微生物和代谢炎症诱导物如何将APPR与动脉粥样硬化和脓毒症联系起来尚待阐明。该提议的中心假设是先天免疫的关键细胞内适配器MyD 88将脓毒症和动脉粥样硬化中的促炎性线索与APPR联系起来。此外，我们假设，在响应促炎症，氧化剂和代谢应激，MyD 88介导的信号诱导核输入的应激反应性转录因子（SRTF），负责遗传重编程的基础APPR。为了验证这两个假设，我们将使用最近设计的新型细胞穿透蛋白，靶向MyD 88，一种位于Toll样/IL 1/IL 18受体附近的衔接蛋白。为了描述APPR中核输入机制的特异性，我们将设计、生产和测试细胞穿透抑制剂，以靶向将SRTFs穿梭至细胞核的输入蛋白（核粒转运蛋白）α 1。使用表现APPR的动物模型，我们将评估这些细胞穿透抑制剂的体内作用机制和治疗潜力。所提出的研究的可行性通过申请人使用细胞穿透蛋白和肽抑制MyD 88-STAT 1/STAT 3-和输入蛋白（核粒细胞转运蛋白）α介导的信号传导途径的最新进展而得到强调。这些进展由定制设计的SRTFs核输入抑制剂证明，该抑制剂有效地纠正了炎症、凋亡、组织损伤和动脉粥样硬化的异常遗传程序。进一步的进展包括SOCS 3的重组细胞穿透形式，其有效地减弱了这些机制中的一些。从拟议的调查结果将有助于利用潜在的细胞穿透蛋白质和肽用于翻译研究，并提供了一个了解的信号中间体，串扰，和细胞内检查点控制APPR。因此，所提出的研究满足了对新的抗炎药物的迫切需要，以抑制APPR及其伴随的心血管系统损伤在脓毒症和动脉粥样硬化。公共卫生相关性：我们建议进一步开发一种创新形式的细胞内蛋白/肽治疗，以消除炎症和重要器官血管损伤。具体来说，我们将研究血液中毒（败血症）和动脉硬化（动脉粥样硬化）的实验模型。在美国，每年有100万例心脏病发作、783，000例中风和估计750，000例住院败血症患者是这些炎症性疾病的基础。

DESCRIPTION (provided by applicant): Inflammation, as the major mechanism of disease, underlies vascular system collapse in sepsis ("septic shock") and the development of atherosclerosis. Both disease processes converge prominently in the 60-85 year age group wherein sepsis-related mortality reaches 45%, as compared to a 30% mortality rate encompassing all age groups. Proinflammatory cytokines induce acute phase protein response (APPR). APPR is manifested by the expression of a biomarker, C-reactive protein (CRP), and by elevated levels of serum amyloid proteins, complement proteins, coagulation factors, and protease inhibitors-each contributing to cardiovascular inflammatory injury. CRP and other biomarkers related to the inflammatory process have been the focus of recent epidemiological and clinical studies. Annually, this process underlies 1 million heart attacks, 783,000 strokes, and an estimated 750,000 patients hospitalized with sepsis in the United States. How a high cholesterol Western diet and other microbial and metabolic inducers of inflammation link APPR to atherosclerosis and sepsis awaits elucidation. The central hypothesis of this proposal is that the key intracellular adapter of innate immunity, MyD88, links proinflammatory cues to APPR in sepsis and atherosclerosis. Furthermore, we hypothesize that in response to proinflammatory, oxidant, and metabolic stress, MyD88-mediated signaling induces nuclear import of stress-responsive transcription factors (SRTFs) that are responsible for the genetic reprogramming underlying APPR. To test these two hypotheses, we will use recently-engineered novel cell-penetrating proteins that target MyD88, an adapter protein positioned proximally to Toll-like/IL1/IL18 receptors. To delineate specificity of nuclear import machinery in APPR, we will design, produce, and test cell-penetrating inhibitors to target importin (karyopherin) alpha 1 that shuttles SRTFs to the nucleus. Using animal models that manifest APPR, we will assess the in vivo mechanism of action and therapeutic potential of these cell-penetrating inhibitors. Feasibility for the proposed study is underscored by the applicant's recent advances to suppress the MyD88-STAT1/STAT3- and importin (karyopherin) alpha-mediated signaling pathways using cell-penetrating proteins and peptides. These advances are evidenced by a custom-designed SRTFs nuclear import inhibitor that effectively corrected the aberrant genetic programs for inflammation, apoptosis, tissue injury, and atherosclerosis. Further advances include a recombinant cell penetrating form of SOCS3 that effectively attenuated some of these mechanisms. Results from the proposed investigation will help to harness the potential of cell-penetrating proteins and peptides for use in translational studies, and provide an understanding of the signaling intermediates, crosstalk, and intracellular checkpoints that control APPR. Thus, the proposed studies meet the urgent need for new anti-inflammatory drugs to suppress APPR and its attendant cardiovascular system injury in sepsis and atherosclerosis. PUBLIC HEALTH RELEVANCE: We propose to further develop an innovative form of intracellular protein/peptide therapy to extinguish inflammation and injury to blood vessels in vital organs. Specifically, we will study experimental models of blood poisoning (sepsis) and hardening of arteries (atherosclerosis). Annually, these inflammation-based diseases underlie 1 million heart attacks, 783,000 strokes, and an estimated 750,000 hospitalized sepsis patients in the United States.