Role of n3 PUFAs and inflammation-resolving lipid mediator, RvD1, in alcoholic liver disease

酒精性肝病（ALD）是美国（特别是肯塔基州）的一个主要且日益严重的健康问题， 国际吧尽管这个问题的严重性，但没有FDA批准的治疗ALD的任何阶段。在 此外，对疾病进展和严重程度的机制和调节因素还不清楚。膳食 脂肪与酒精消耗在ALD发病机制中起着重要的相互作用，然而，n3 ALD中的PUFA没有明确定义。我们的中心假设是n3 PUFA在ALD中是有益的，部分是通过 n3-PUFA衍生的促消退介质，促进炎症消退，改善肠道-肝脏 轴，以及随后的肝损伤衰减。我们认为消退素D1（Resolvin D1，RvD 1）是一种有效的治疗药物， 通过RvD 1-FPR 2-NEAT 1信号传导抑制促炎性细胞因子并 促进肝细胞损伤的修复，部分通过增强促修复巨噬细胞。我们 假设由于受损的RvD 1产生/信号传导而导致的炎症消退受损是导致炎症消退的原因之一。 关键的营养因素的进展ALD和酒精性肝炎（AH）的严重程度在人类。 该提案的具体目标是：目标1。为了测试n3 PUFA是否对EtOH产生有益影响， 通过增强炎症消退的有效性和通过 通过增加n3-PUFA衍生的特化促消退介质修复肝细胞损伤 促进 （SPM），以及RvD 1-FPR 2和Neat 1介导的促炎细胞因子信号传导的抑制， 将促炎巨噬细胞重编程为促恢复表型。野生型（WT），Fpr 2-/-， neat 1-/-和转基因fat-1小鼠（能够内源性地将n6转化为n3 PUFA）将用于本研究。 瞄准我们还将研究RvD 1的治疗效果，利用一种新的纳米颗粒技术， 靶向RvD 1递送检查n3-PUFA的作用 植物来源的可食用外泌体注入肝脏目标2.到 和RvD 1在维持肠道屏障完整性和在实验性ALD中的肠道炎症的解决中的作用。 我们将：i）在体内、动物模型和体外、肠类器官培养中测试n3 PUFA或RvD 1 通过减弱肠免疫失调改善肠屏障损伤; ii）在体内测试是否 RvD 1-FPR 2轴的破坏加剧，而阻断Neat 1信号转导减弱肠道炎症 和肠道屏障完整性的改变; iii）测试施用工程化的 细菌菌株在肠中将n6转化为n3 PUFA，从而减弱小鼠的肠屏障功能障碍。 在目标3中，我们试图将我们在动物模型中的发现转化并扩展到人类ALD。利用去身份化 i）确定n3-PUFA膳食补充剂对血浆SPM水平、标志物 肝损伤，全身炎症，和肠道通透性在大量饮酒的个人; ii）检查血浆 从AH获得的全血和外周血单核细胞（PBMC）中的SPM水平和SPM合成 iii）测试RvDl是否改善从AH患者获得的PBMC的吞噬作用/吞噬作用。

Alcoholic liver disease (ALD) is a major and increasing health problem in the US (especially in Kentucky) and worldwide. In spite of the magnitude of this problem, there is no FDA-approved therapy for any stage of ALD. In addition, the mechanisms and regulators of the disease progression and severity are not well understood. Dietary fats play an important interactive role with alcohol consumption in ALD pathogenesis, however, the role of n3 PUFAs in ALD are not well defined. Our central hypothesis is that n3 PUFAs are beneficial in ALD, in part, via n3-PUFA-derived pro-resolving mediators which facilitate inflammation resolution, improvement in the gut-liver axis, and subsequent attenuation of liver injury. We propose that resolvin D1 (RvD1) is a potent therapeutic agent in severe ALD acting via RvD1-FPR2-NEAT1 signaling to suppress pro-inflammatory cytokines and to promote repair of hepatocellular damage, in part, via enhancement of pro-restorative macrophages. We postulate that compromised inflammation resolution due to impaired RvD1 production/signaling is one of the critical nutritional contributing factors to the progressive ALD and severity of alcoholic hepatitis (AH) in humans. The Specific Aims of the proposal are: Aim 1. To test whether n3 PUFAs exert beneficial effects on EtOH- associated liver injury/inflammation by enhancing the effectiveness of inflammation resolution and by repair of hepatocellular damage through increase in n3-PUFA-derived specialized pro-resolving mediators promoting (SPMs), and RvD1-FPR2 and Neat1-mediated suppression of pro-inflammatory cytokine signaling and reprogramming pro-inflammatory macrophages into a pro-restorative phenotype. Wild Type (WT), Fpr2-/-, Neat1-/-, and transgenic fat-1 mice (which are able to endogenously convert n6 to n3 PUFAs) will be used in this Aim. We will also examine the therapeutic effectiveness of RvD1 utilizing a novel nanoparticle technology of targeted RvD1 delivery examine the role n3-PUFAs to the liver with plant-derived edible exosomes. Aim 2. To and RvD1 in maintaining gut barrier integrity, and in the resolution of intestinal inflammation in experimental ALD. We will: i) test in vivo, in animal models, and in vitro, in intestinal organoid culture, whether n3 PUFA or RvD1 improve intestinal barrier damage by attenuating intestinal immune dysregulation; ii) test in vivo whether disruption of the RvD1-FPR2 axis exacerbates, while blocking Neat1 signaling attenuates intestinal inflammation and alterations in the gut barrier integrity; iii) test a novel therapeutic strategy of administering an engineered bacteria strain to convert n6 to n3 PUFAs in the intestine and thus to attenuate gut barrier dysfunction in mice. In Aim 3, we seek to translate and extend our findings in animal models to human ALD. Utilizing de-identified human samples we will: i) determine effects of n3-PUFA dietary supplementation on plasma SPM levels, markers of liver injury, systemic inflammation, and intestinal permeability in heavy drinking individuals; ii) examine plasma SPM levels, and SPM synthesis in whole blood and peripheral blood monocytes (PBMCs) obtained from AH patients; and iii) test whether RvD1 improves phagocytosis/efferocytosis of PBMCs obtained from AH patients.