Nanoparticle-mediated targeting of hepatic macrophages to mitigate inflammation in alcoholic liver disease

摘要 酒精滥用造成的持续肝损伤导致的慢性和未解决的肝脏炎症可能导致纤维化， 肝硬变和最终的肝细胞癌--这是与癌症相关的快速增长的原因 世界上最大的死亡率。美国有近450万成年人患有慢性肝病。有效 由于穷人的原因，针对肝脏炎症的安全治疗策略仍然是一个未得到满足的临床需求 当前治疗方法的药代动力学、毒性和缺乏特异性。纳米粒(NP)介导靶向药物 给药可达到较高的肝脏浓度和较低的全身浓度。我们有 证明NPs可以靶向G蛋白偶联的胆汁酸受体1(Gpbar1)。 库普弗细胞，调节肝脏炎症反应。用pH响应型聚合物包覆纳米颗粒 进一步控制药物释放动力学，促进包囊药物释放到 酸性炎症微环境。因此，这个R21项目的首要目标是开发Kupffer 可用于靶向刺激Gpbar1和Gpbar1的细胞靶向双功能NP制剂 同时释放pH响应的抗炎治疗药物。我们假设联合行动 Gpbar1的刺激和抗炎治疗在缓解慢性肝脏方面将具有相加的作用 与酒精性肝病相关的炎症。我们提出的目标是：目标1：在体内评估 靶向Gpbar1的NPs的靶向能力和生物分布。我们将研究和优化目标 INT-777修饰的可生物降解聚合物纳米粒子的性能和生物分布 Gpbar1激动剂，使用广泛使用的慢性+酗酒的酒精性肝病动物模型。目标2：实现 考察所设计的纳米粒在体内外的治疗效果。基于优化后的性能 在目标1中，将对NPs进行进一步修饰，以加入抗炎药物地塞米松，并进行测试 对照酒精处理的肝脏芯片模型和慢性+暴饮暴食的动物模型，确定它们的 药代动力学和药效学(PK/PD)特性。我们将使用组织学，生物标记物分析，胶原蛋白 检测和细胞因子ELISA研究我们的双功能纳米粒能否显著缓解慢性肝病 炎症与不含INT-777的免费药物或NPs相比。我们的结果将导致 开发和测试慢性肝炎的新治疗策略，在紧急情况下 需要开发和安全、一贯有效的治疗方法。

ABSTRACT Chronic and unresolved liver inflammation due to persistent liver injury from alcohol abuse can lead to fibrosis, cirrhosis and eventually hepatocellular carcinoma (HCC) – which is the fasted growing cause of cancer-related mortality in the world. Nearly 4.5 million adults in the United States are living with chronic liver disease. Effective and safe treatment strategies against hepatic inflammation remains an unmet clinical need due to the poor pharmacokinetics, toxicity and lack of specificity of current therapies. Nanoparticle (NP)-mediated targeted drug delivery can achieve high hepatic concentrations and low systemic concentrations of the drug. We have demonstrated that NPs can be targeted towards G-protein coupled bile acid receptor1 (Gpbar1) expressed by Kupffer cells, to regulate the hepatic inflammatory response. Coating the NPs with pH-responsive polymers offers further control over the drug release kinetics by facilitating release of the encapsulated drugs into the acidic inflammatory microenvironment. Therefore, the overarching goal of this R21 project is to develop a Kupffer cell-targeting dual-functional NP formulation that can be used for targeted stimulation of Gpbar1 and simultaneous pH-responsive release of anti-inflammatory therapeutics. We hypothesize that the combined action of Gpbar1 stimulation and anti-inflammatory therapy will have an additive effect in mitigating chronic liver inflammation associated with alcoholic liver disease. Our proposed aims are: Aim 1: To evaluate the in vivo targeting capabilities and biodistribution of the Gpbar1-targeted NPs. We will study and optimize the targeting capabilities and biodistribution of our biodegradable polymeric NPs surface decorated with INT-777- a potent Gpbar1 agonist, using a widely used chronic-plus-binge animal model of alcoholic liver disease. Aim 2: To investigate the therapeutic efficacy of the designed NPs in vitro and in vivo. Based on the optimized properties in Aim 1, the NPs will be further modified to incorporate the anti-inflammatory drug dexamethasone, and tested against alcohol-treated liver-chip models and the chronic-plus-binge animal models to determine their pharmacokinetic and pharmacodynamic (PK/PD) properties. We will use histology, biomarker analysis, collagen assays and cytokine ELISA to study whether our dual-functional NPs can significantly mitigate chronic hepatic inflammation compared to free drugs or NPs without INT-777. Our results will lead to a paradigm shift in the development and testing of new therapeutic strategies for chronic liver inflammation, where there is an urgent need for the development and safe and consistently effective therapies.