Molecular Mechanisms Of Post-Transplant Recurrent Alcoholic Liver Disease

项目总结 这项提案概述了一项研究神经酰胺在酒精诱导的脂滴中的作用的计划。 生物发生和肝移植后酒精性肝病(ALD)复发。这项研究将使用小说在 体外和体内模型系统研究神经酰胺合成酶神经酰胺的假说 合成酶6(CerS6)通过转录调控调节酒精诱导的脂滴的生物发生 脂滴蛋白Perilipin 2(PLIN2)。此外，这项提案将调查两者之间的关联 CerS6在酒精性脂肪变性和移植后复发ALD移植物丢失中的上调。研究人员的目标是 1)确定CerS6调节PLIN2和脂滴生物发生的机制；2) 肝细胞特异性CerS6消融术对大鼠血脂和血糖动态平衡的影响 实验性ALD和3)确定CerS6表达是否预测移植后移植物丢失 复发性ALD。 为了实现这些特定的目标，研究人员建议在基因上使用CRISPR/Cas9 转基因小鼠和人类乙醇代谢VL17A细胞进行综合代谢 表型分析、类脂组学分析、生化和功能分析。此外，研究人员的目标是 检测CerS6免疫组织化学表达是否可预测ALD术后复发患者的移植物丢失 移植。该项目的结果将为将CerS6作为诊断和治疗的靶点提供证据 ALD移植后复发的生物标志物。

PROJECT SUMMARY This proposal outlines a plan to investigate the role of ceramides in alcohol-induced lipid droplet biogenesis and in post-liver transplant alcoholic liver disease (ALD) recurrence. The research will use novel in vitro and in vivo model systems to investigate the hypothesis that the ceramide synthetic enzyme ceramide synthase 6 (CerS6) regulates alcohol-induced lipid droplet biogenesis through the transcriptional regulation of the lipid droplet protein, Perilipin 2 (PLIN2). Furthermore, this proposal will investigate the association between CerS6 upregulation in alcoholic steatosis and graft loss in post-transplant recurrent ALD. The researcher aims 1) to determine the mechanisms by which CerS6 regulates PLIN2 and lipid droplet biogenesis; 2) to determine the effects of hepatocyte-specific CerS6 ablation on lipid and glucose homeostasis in experimental ALD and 3) to determine if CerS6 expression predicts graft loss in post-transplant recurrent ALD. To accomplish these Specific Aims, the researcher proposes to use CRISPR/Cas9 genetically engineered mice and human ethanol-metabolizing VL17A cells to perform comprehensive metabolic phenotyping, lipidomics analyses, and biochemical and functional assays. In addition, the researcher aims to examine if CerS6 immunohistochemical expression predicts graft loss in patients with recurrent ALD post- transplant. Results of this project will provide evidence for targeting CerS6 as a diagnostic and therapeutic biomarker in post-transplant ALD recurrence.