Role of Nlrp3 inflammasome signaling in alcoholic liver disease progression

酒精性肝病（ALD）是当今西方社会的主要疾病负担之一。虽然近年来其患病率有所增加，但疾病进展为严重肝脏炎症，肝纤维化和肝细胞癌仍然不清楚。含有3个炎性体的多蛋白质复合物NLR家族pyrin结构域涉及促进疾病进展、促进细胞炎症并有助于肝星状细胞的分化，从而加速纤维化的发展。这些研究促进了新型Nlrp3炎性体干扰化合物的开发，该化合物刚刚进入第一个用于治疗非酒精性脂肪肝的人体研究。在先前发表的研究中，我们证明，Nlrp3炎性体过度激活加剧了小鼠肝脏炎症和肝纤维化的发展。在初步实验中，我们发现IL-18是肝星状细胞活化的驱动因素，酒精孵育处理增加了肝细胞IL-18的产生。我们还分析了两个独立的肝细胞癌患者队列中炎性小体级联相关的SNP，总发病率为10%。从逻辑上讲，我们提出了这样的假设，即细胞特异性Nlrp3炎性体激活是酒精性肝病中触发肝脏炎症和肝纤维化的中心机制。三个工作包将患者数据的评估与小鼠模型中的体内实验联系起来，并推进细胞培养和类器官系统。对于后者，我们将整合Soumita Das教授（加州大学圣地亚哥分校）的知识和专业知识，他是类器官系统的专家，在Charité柏林我们不断发展的肝脏研究部门。我们将采用多细胞型共培养与细胞缺陷或广泛精通Nlrp3信号。因此，我们的体内实验类似地集中于Nlrp3或IL-18的细胞特异性激活或整体缺失。将饮用水中的酒精与喂给小鼠8周的西式饮食相结合，将允许更密切地描绘单个疾病组分及其在炎症信号传导中的作用。更具体地说，我们将采用多色流式细胞术分析单个炎症细胞亚群，以预测它们在疾病进展和治疗干预中的作用。最后，我们与Andreas海因茨教授（最近成立的合作研究中心Transregio 265-失去和恢复对药物摄入的控制的发言人）Charité柏林建立了一项合作努力，以评估（ALD）进展的全谱。在这一队列中，我们将评估多种标志物，包括细胞因子、免疫细胞分布和微小RNA表达。随着全基因组测序的进行，我们将有独特的机会发现ALD患者的炎性级联改变。

Alcoholic liver disease (ALD) is one of the major disease burdens in western society of our time. Although its prevalence has increased over recent years, disease progression towards severe liver inflammation, liver fibrosis and hepatocellular carcinoma are still not understood. The multi-protein complex NLR family pyrin domain containing 3 inflammasome has been implicated in the promotion of disease progression, boosting cellular inflammation and contributing to differentiation of hepatic stellate cells thereby accelerating the development of fibrosis. These studies fostered the development of novel Nlrp3 inflammasome-interfering compounds, which just entered in the first human studies for the treatment of non-alcoholic fatty liver disease. In previous published studies we demonstrate, that Nlrp3 inflammasome overactivation aggravates the development of liver inflammation and liver fibrosis in mice. In preliminary experiments, we discovered that IL-18 is a driver of hepatic stellate cell activation and alcohol incubation treatment increased hepatocellular IL-18 production. We also analyzed inflammasome-cascade related SNPs in two independent cohorts of patients with hepatocellular cancer and faced a total incidence of 10%. Logically, we propose the HYPOTHESIS that cell-specific Nlrp3 inflammasome activation is a central mechanism that triggers hepatic inflammation and liver fibrogenesis in alcoholic liver disease. Three work packages link evaluation of patient data with in vivo experiments in murine models and advance cell culture and organoid systems. For the latter we will integrate the knowledge and expertise of Prof. Soumita Das (University of California, San Diego), an expert in organoid systems, in our growing liver research unit at Charité Berlin. We will employ multi-celltype coculture with cells either deficient or extensively proficient in Nlrp3 signaling. Consequently, our in vivo experiments are similarly focused on cell specific activation or global deletion of Nlrp3 or IL-18. Combining alcohol in drinking water alongside a Western style diet fed to mice for 8 weeks, will allow a closer delineation of the individual disease components and their role in inflammatory signaling. More specifically, we will employ multi-colour flow cytometry to analyze the individual inflammatory cell subsets in order to project their role in disease progression and therapeutic interventions. Finally, we have established a collaborative effort with Prof. Andreas Heinz (spokesperson of the recently established Collaborative Research Center Transregio 265 – Losing and Regaining Control over Drug Intake) Charité Berlin to evaluate the full spectrum of (ALD) progression. In this cohort of we will evaluate a multiplicity of markers including cytokines, immune cell distribution and microRNA expression. As whole genome sequencing will be performed we will have the unique opportunity to uncover inflammasome cascade alteration in patients with ALD.