Intracellular Therapeutics for Inflammatory Liver Injury

描述(由申请人提供)：酒精性肝病(ALD)在美国估计有200万患者，4年期间死亡率高达65%。我们假设，急性或慢性酒精滥用改变了细胞内促炎信号的中介物和抑制物之间的动态平衡。这种信号在肝细胞的遗传重编程中达到顶峰，表现为促炎症和促凋亡介质的表达。我们将研究在T细胞介导的肝炎模型中，乙醇如何改变两个关键的细胞内介质，信号转导和转录激活因子(STAT)1和3的相反作用。酒精诱导的由STAT1介导的潜在有害(促凋亡)信号和由STATS介导的有益(抗凋亡)信号之间的失衡将被我们称为细胞内蛋白治疗的新策略所研究，以靶向过度激活STAT1导致的促炎和促凋亡信号。我们将把我们的成功设计和体内交付的细胞因子信号生理抑制物扩展到胞浆蛋白，该蛋白可消除巨噬细胞(Kupffer细胞)中的促炎信号，后者在酒精相关的炎症性肝损伤中发挥重要作用。此外，我们还开发了一种穿透性核进口抑制肽，作为一类新型抗炎和抗凋亡药物的原型，在动物模型中抑制促炎细胞因子/趋化因子的产生，防止T细胞或巨噬细胞介导的大量肝细胞凋亡/坏死，并降低死亡率。由于这种细胞穿透性多肽抑制剂针对的是负责应激反应转录因子核输入的细胞内适配器，我们将分析核输入适配器在酒精诱导的肝细胞损伤中的作用。总而言之，我们预计从这些研究中获得的新知识将推动我们的细胞内蛋白质疗法的概念，以消除酒精加剧的肝脏炎症和细胞凋亡，并有助于为美国约200万ALD患者中快速衰竭的肝脏开发新的治疗方法。根据总体工作范围，本拨款申请是根据PA-05-074《酒精诱导组织损伤的机制》提交的。

DESCRIPTION (provided by applicant): Alcoholic liver disease (ALD) afflicts an estimated 2 million patients in the US with an astounding 65% mortality rate over a 4 year period. We hypothesize that acute or chronic alcohol abuse alters homeostatic balance that exists between intracellular mediators and suppressors of proinflammatory signaling. This signaling culminates in genetic reprogramming of liver cells manifested by expression of proinflammatory and proapoptotic mediators. We will study how ethanol alters the opposing roles of two key intracellular mediators, signal transducer and activator of transcription (STAT) 1 and 3 in T cell-mediated hepatitis model. The alcohol-induced imbalance between potentially harmful (proapoptotic) signaling mediated by STAT1 and beneficial (anti-apoptotic) signaling mediated by STATS will be studied using our new strategy termed intracellular protein therapy to target proinflammatory and proapoptotic signaling resulting from excessive STAT1 activation. We will expand our successful design and in vivo delivery of physiologic suppressors of cytokine signaling to the cytoplasmic protein known to ablate proinflammatory signaling in macrophages (Kupffer cells) which play a significant role in alcohol-associated inflammatory liver injury. Moreover, we have developed a cell-penetrating nuclear import inhibitory peptide as a prototype of a new class of anti- inflammatory and anti-apoptotic agents which suppresses production of proinflammatory cytokines/chemokines, prevents massive liver apoptosis/necrosis mediated by T cells or macrophages, and reduces mortality rates in animal models. Since this cell-penetrating peptide inhibitor targets intracellular adaptors responsible for nuclear import of stress responsive transcription factors, we will analyze the role of nuclear import adaptors in alcohol-induced injury of liver cells. Cumulatively, we anticipate that the new knowledge gained from these studies will, advance our concept of intracellular protein therapy to extinguish alcohol-exacerbated liver inflammation and apoptosis, and contribute to the development of new therapies for the rapidly failing liver among an estimated 2 million ALD patients in the US. Based on the overall workscope, this grant application is submitted in response to PA-05-074 "Mechanisms of Alcohol-Induced Tissue Injury".