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

肝脏富集转录因子作为酒精性肝炎的预后标志物和治疗靶点

• 批准号: 10428560
• 负责人: Gavin E Arteel
• 金额: $ 24.1万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-22 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10428560
• 关键词: Alcoholic Hepatitis; Alcoholic Liver Diseases; Area; Biological Markers; CSF3 gene; Cell Differentiation process; Cells; Clinical; Clinical Data; Coupled; DNA Binding; Data; Development; Disease; Down-Regulation; EGF gene; Enrollment; Epidermal Growth Factor Receptor; Failure; Functional disorder; Genes; Glycoproteins; Goals; HNF4A gene; Hepatobiliary; Hepatocyte; Human; Immunoprecipitation; In Vitro; Inflammation; Inflammatory; Knowledge; Lead; Ligands; Link; Liver; Liver Fibrosis; Mass Spectrum Analysis; Mediating; Microdissection; Molecular; Natural regeneration; Neutrophil Infiltration; Organoids; Outcome; Pathway interactions; Patient-Focused Outcomes; Patients; Peripheral; Phenotype; Play; Post Translational Modification Analysis; Prognostic Marker; Protein Isoforms; Proteins; Proteome; Proteomics; Reaction; Regenerative response; Regulation; Reporter; Research; Role; Sampling; Serum; Serum Proteins; Severities; Severity of illness; Signal Transduction; System; Techniques; Therapeutic; Tissues; base; cohort; effective therapy; experimental study; genetic signature; genomic profiles; in vitro Model; innovation; liver function; mortality; novel; novel therapeutic intervention; novel therapeutics; precision medicine; prednisolone; premature; preservation; promoter; protein biomarkers; response; screening; standard of care; stem cells; targeted treatment; therapeutic target; tool; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; treatment response; Alcoholic Hepatitis; Alcoholic Liver Diseases; Area; Biological Markers; CSF3 gene; Cell Differentiation process; Cells; Clinical; Clinical Data; Coupled; DNA Binding; Data; Development; Disease; Down-Regulation; EGF gene; Enrollment; Epidermal Growth Factor Receptor; Failure; Functional disorder; Genes; Glycoproteins; Goals; HNF4A gene; Hepatobiliary; Hepatocyte; Human; Immunoprecipitation; In Vitro; Inflammation; Inflammatory; Knowledge; Lead; Ligands; Link; Liver; Liver Fibrosis; Mass Spectrum Analysis; Mediating; Microdissection; Molecular; Natural regeneration; Neutrophil Infiltration; Organoids; Outcome; Pathway interactions; Patient-Focused Outcomes; Patients; Peripheral; Phenotype; Play; Post Translational Modification Analysis; Prognostic Marker; Protein Isoforms; Proteins; Proteome; Proteomics; Reaction; Regenerative response; Regulation; Reporter; Research; Role; Sampling; Serum; Serum Proteins; Severities; Severity of illness; Signal Transduction; System; Techniques; Therapeutic; Tissues; base; cohort; effective therapy; experimental study; genetic signature; genomic profiles; in vitro Model; innovation; liver function; mortality; novel; novel therapeutic intervention; novel therapeutics; precision medicine; prednisolone; premature; preservation; promoter; protein biomarkers; response; screening; standard of care; stem cells; targeted treatment; therapeutic target; tool; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; treatment response

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.

电池 - UO1（RFA AA-18-003） 抽象的 肝细胞衰竭是酒精性肝炎患者（AH）的标志性发现。当前的疗法 不完全有效，需要有针对性的疗法。大多数研究都集中在 炎症的作用。相反，肝细胞衰竭的基础机制和 随后在AH中的再生反应较差。我们的小组表明严重的AH 具有大量但无效的导管细胞积累的特征。中心目的 该建议是确定肝细胞衰竭和 随后在AH中的再生反应差。最终目标是识别预后 标记和治疗靶标可用于精密医学。我们已经产生了强大 人类生活中的初步数据显示：（1）ALD早期形式的进展为AH是 其特征是HNF4A的功能下降。 （2）HNF4A网络足迹 与AH患者的疾病严重程度密切相关。 （3）TGFβ1和EGF是上游的关键 AH中的调节剂并调节培养的人肝细胞中的LETF。 （4） AH中的导管反应显示富集的炎症，纤维化和增殖信号，以及 HNF4A活性降低。 （5）由左目标基因编码的分泌蛋白是 在AH患者的血清中检测到。基于这些强大的初步数据，我们的具体目的 （1）揭示了通过TGFβ1调节HNF4A同工型的分子机制， 通过MS耦合表征HNF4A P1依赖同工型的相互作用组 免疫沉淀。此外，我们旨在识别HNF4A P2的DNA结合区域 chipseq的同工型，并产生启动子报告系统，该系统将用于体外高级 新型药物的水槽筛查。 （2）研究的相关性和基因组概况 通过RNASEQ的RNASEQ的凝结增生和脱离分化的肝细胞 微分解区域以及基因特征与临床结果的相关性。我们有 发达的人体器官的凝管反应，代表了一种研究效果的新工具 HNF4A中的TGFβ1和EGFR配体的。 （3）。确定新的血清蛋白生物标志物 AH通过糖蛋白选择偶联来研究血清N连接的糖蛋白室 通过Snap-shot通过分析了血清蛋白质组的翻译后修饰 蛋白质组学阵列。我们将将血清蛋白质组学数据的分析与转录组学研究结合 总和显微解剖的肝组织，并将所得的特征与临床结局相关联 包括死亡率和对泼尼松龙和G-CSF的反应。