Role of Soluble Epoxide Hydrolase in Alcohol-Associated Liver Disease

酒精相关性肝病(ALD)是一系列肝脏疾病，从肝脏脂肪变性到 脂肪性肝炎伴不同程度的纤维化和肝硬变。肌萎缩侧索硬化症是发病率、死亡率和 美国和世界各地的医疗成本。然而，目前还没有FDA批准的治疗ALD任何阶段的方法。 对疾病进展的机制和媒介的了解也不完全 严肃性。可溶性环氧化物水解酶(S-EH)，调节内源激素代谢的主酶 生物活性脂类(如环氧脂肪酸，EP-FAs)最近被认为是一个新兴的治疗靶点 在多种疾病中。该项目的总体目标是测试S-EH抑制剂的治疗效果 不同阶段的酒精性肝病的严重程度，并为将S-高血压抑制作为新药提供机制基础 酒精性肝病的治疗。目的1.检测S抑制EH的疗效 作为ALD的一种新的治疗策略。我们将：i)测试药物对S-EH活性的调节作用 抑制或S-EH(EPHX2)基因消融可有效减轻或预防乙醇诱导的小鼠肝损伤 实验性ALD；ii)评价S-EH抑制是否导致血液和肝脏EP-FAs的稳定；以及iii) EP-FA水平变化与肝损伤标志物的相关性。系统性、靶向性地给药S-- EH抑制剂(t-TUCB和TPPU)以及WT和EPHX2-/-小鼠(全局和肝脏特异性)将用于多个 ALD的动物模型，产生不同的疾病严重程度阶段，并概括不同的 人类阿尔茨海默病的特征。这使得可以严格评估S-EH在不同阶段的抑制效果 严重的阿尔茨海默病。将采用治疗和预防模式。目的2.探索机制(S) S-EH抑制对实验性阿尔茨海默病的有益作用。我们将确定EP-FA是否 S-EH抑制介导的稳定作用在减轻乙醇诱导的肝损伤中起关键作用。我们会 测试n3和n6 EP-FA是否通过增强M2巨噬细胞极化而发挥更大的有益作用， 通过EP-FAs-PPARγ-CXCL1介导增加促进修复/促进分解的巨噬细胞表型 中性粒细胞浸润减少。S-EH抑制剂，WT，转基因FAT-1小鼠(内源性转化n6 多不饱和脂肪酸到n3多不饱和脂肪酸)，以及肝细胞特异性多不饱和脂肪酸受体γ-/-小鼠。体内和体外研究将是 已执行。目的3.观察乙醇对人酒精性肝病S-EH和EP-FAs的影响。利用去离子化 经鉴定的人血浆和全血样本，我们将：i)评估血浆EP-FAs和 建立酒精中毒患者肝损伤生物标志物与全身炎症的关系 相关性肝炎(AH)；II)确定饮食补充n3-PUFA对血浆EP-FAs的影响 检测S-EH抑制和EP-FA(包括n3和n6 EP-FA) 能有效降低基础和内毒素刺激的全血促炎症细胞因子的产生 采集急性肝炎患者外周血单核细胞。

Alcohol-associated liver disease (ALD) is a spectrum of liver disorders ranging from hepatic steatosis to steatohepatitis with varying degrees of fibrosis and cirrhosis. ALD is a major cause of morbidity, mortality, and health care costs in the US and worldwide. However, there is no FDA-approved therapy for any stage of ALD. There is also an incomplete understanding of the mechanisms and mediators of disease progression and severity. Soluble epoxide hydrolase (s-EH), a master enzyme that regulates the metabolism of endogenous bioactive lipids (e.g., epoxy-fatty acids, Ep-FAs), has recently been recognized as an emerging therapeutic target in multiple diseases. The overall goal of this project is to test the therapeutic efficacy of s-EH inhibition at different stages of ALD severity, and to provide a mechanistic foundation for using s-EH inhibition as a novel therapy for alcohol-induced liver pathology. Aim 1. To test the therapeutic effectiveness of s-EH inhibition as a novel therapeutic strategy for ALD. We will: i) test whether modulation of s-EH activity by pharmacological inhibition or s-EH (Ephx2) genetic ablation can effectively attenuate or prevent EtOH-induced liver injury in experimental ALD; ii) evaluate whether s-EH inhibition leads to stabilization of blood and liver Ep-FAs; and iii) correlate changes in Ep-FA levels with markers of liver injury. Systemic and targeted liver-specific delivery of s- EH inhibitors (t-TUCB and TPPU), and WT and Ephx2 -/- mice (global and liver-specific) will be used in multiple animal models of ALD, which produce different stages of disease severity and which recapitulate different features of human ALD. This allows for a rigorous evaluation of the effects of s-EH inhibition at different stages of ALD severity. Treatment and prevention paradigms will be applied. Aim 2. To explore mechanism(s) underlying the beneficial effects of s-EH inhibition in experimental ALD. We will determine whether Ep-FA stabilization mediated by s-EH inhibition plays a critical role in attenuation of EtOH-induced liver injury. We will test whether n3 vs n6 Ep-FAs exert a greater beneficial effect by enhancing M2 macrophage polarization, increasing a pro-restorative/pro-resolving macrophage phenotype, and by Ep-FAs-PPARγ-CXCL1-mediated reduction of neutrophil infiltration. s-EH inhibitors, WT, transgenic fat-1 mice (which endogenously convert n6 PUFAs to n3 PUFAs), and hepatocyte-specific Pparγ-/- mice will be used. In vivo and in vitro studies will be performed. Aim 3. To evaluate EtOH-induced alterations in s-EH and Ep-FAs in human ALD. Utilizing de- identified human plasma and whole blood samples, we will: i) evaluate alterations in plasma Ep-FAs and establish relationships between biomarkers of liver injury and systemic inflammation in patients with alcohol- associated hepatitis (AH); ii) determine the effects of n3-PUFA dietary supplementation on plasma Ep-FAs in heavy drinking individuals; and iii) test whether s-EH inhibition and Ep-FAs (including both n3 and n6 Ep-FAs) can effectively decrease basal and LPS-stimulated pro-inflammatory cytokine production in whole blood and peripheral blood monocytes obtained from AH patients.