Liver-enriched Transcription Factors as Prognostic Markers and Therapeutic Targets in Alcoholic Hepatitis

巴特勒-UO1(RFA AA-18-003) 摘要 肝细胞衰竭是酒精性肝炎(AH)患者的标志性发现。现代疗法 并不是完全有效，需要有针对性的治疗。大多数研究都集中在 炎症的作用。相比之下，肝细胞衰竭的机制和 在AH中随后的不良再生反应尚不清楚。我们的小组显示，严重的AH 以大量但低效的导管细胞堆积为特征。的中心目标是 这项建议是为了确定肝细胞衰竭的分子机制和 随后在AH中再生反应差。最终的目标是确定预后 对精准医学有用的标志物和治疗靶点。我们已经产生了强大的 人类肝脏的初步数据显示：(1)ALD的早期形式进展为AH 以HNF4a功能的显著下降为特征。(2)HNF4a网络足迹 与急性肝炎患者的病情严重程度密切相关。(3)转化生长因子β-1和表皮生长因子是关键的上游。 AH中的调节剂和调节培养的人肝细胞中的LETF。(4)基因签名 AH的导管反应表现为丰富的炎性、纤维化和增殖性信号，并 HNF4a活性降低。(5)左靶基因编码的分泌蛋白为 在急性肝炎患者的血清中检测到。基于这些强劲的初步数据，我们的具体目标 目的：(1)揭示转化生长因子β1对HNF4a亚型调控的分子机制。 HNF4a P1依赖异构体相互作用组的MS联用研究 免疫沉淀。此外，我们的目标是确定HNF4a P2的DNA结合区 并构建了一个启动子报告系统，该系统将用于体外高效表达。 新药产量筛选。(2)研究基因的相关性和基因组图谱 AH型胆管细胞增殖和去分化肝细胞的RNA序列分析 显微解剖区域和基因特征与临床结果的相关性。我们有 开发的人类导管反应类器官代表了一种研究这一效应的新工具 转化生长因子β1及其配体在HNF4a中的表达。和(3)。寻找新的血清蛋白生物标志物 糖蛋白选择偶联研究血清N-连接糖蛋白区段的研究 通过快照分析血清蛋白质组的翻译后修饰 蛋白质组阵列。我们将结合血清蛋白质组数据的分析和转录组研究 对肝组织进行整体和显微解剖，并将结果与临床结果相关联 包括死亡率和对强的松龙和G-CSF的反应。

Bataller – UO1 (RFA AA-18-003) ABSTRACT Hepatocellular failure is a hallmark finding in patients with alcoholic hepatitis (AH). Current therapy is not fully effective and targeted therapies are needed. Most research has been focused on the role of inflammation. In contrast, the mechanisms underlying hepatocellular failure and the subsequent poor regenerative response in AH are unknown. Our group showed that severe AH is characterized by a massive, yet inefficient accumulation of ductular cells. The central aim of this proposal is to identify the molecular mechanisms underlying the hepatocellular failure and the subsequent poor regenerative response in AH. The ultimate goal is the identification of prognostic markers and therapeutic targets useful for precision medicine. We have generated strong preliminary data in human livers showing: (1) progression of early forms of ALD to AH is characterized by a profound decrease in the function of HNF4A. (2) HNF4A network footprint closely correlates with disease severity in AH patients. (3) TGFβ1 and EGF are key upstream modulators in AH and regulate LETF in cultured human hepatocytes. (4) The gene signature of the ductular reaction in AH shows enriched inflammatory, fibrogenic and proliferative signals and decreased HNF4A activity. And (5) Secreted proteins encoded by LEFT-target genes are detected in sera of patients with AH. Based on these strong preliminary data, our specific aims are (1) To uncover the molecular mechanism of HNF4A isoform regulation by TGFβ1, by characterizing the interactome of the HNF4A P1 dependent isoforms by MS coupled Immunoprecipitation. Moreover, we aim at identifying the DNA binding regions of HNF4A P2 isoform by ChIPseq, and generating a promoter reporter system that will be used for in vitro high- troughput screening of novel drugs. (2) To investigate the relevance and genomic profile of ductular proliferation and de-differentiated hepatocytes in AH by means of RNAseq of microdissected areas and correlation of gene signatures with clinical outcome. We have developed human organoids of ductular reaction that represents a novel tool to study the effect of TGFβ1 and EGFR ligands in HNF4A. And (3). To identify new serum protein biomarkers of AH by investigating serum N-linked glycoprotein compartment by glycoprotein selection coupled to MS and by analyzing post-translational modifications of the serum proteome by Snap-shot proteomic array. We will combine analysis of serum proteomic data with transcriptomic studies of total and microdissected liver tissue and correlate the resulting signatures with clinical outcomes including mortality and response to prednisolone and G-CSF.