MATRIX-INDUCED REGULATION OF HEPATOCYTE DIFFERENTIATION

基质诱导的肝细胞分化调节

• 批准号: 6524631
• 负责人: AARON W BELL
• 金额: $ 4.62万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-04 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6524631
• 关键词: biological signal transduction; cell differentiation; cell growth regulation; extracellular matrix; laboratory mouse; laboratory rat; liver cells; liver regeneration; messenger RNA; postdoctoral investigator; protein structure function; transcription factor

A cells fate as well as its differentiative state and phenotype are highly dependent on signals from the extracellular matrix. The liver, and hepatocyte cultures in particular, represent and appropriate and convenient model system to study cellular growth and differentiation since several signaling pathways and liver-enriched transcription factors have been identified which acre essential to initiation and maintenance of liver differentia6on/development. Hepatic growth and differentiation is mediated primarily by the combinatorial actions of liver-enriched transcription factors, foremost of which is HNF-4.. To more clearly understand the molecular mechanisms by which extracellular matrix proteins regulate growth, regeneration and differentiation of hepatocytes, this study aims to investigate the matrixspecific alterations in HNF-4 expression, DNA binding, and transcriptional activity in differentiating hepatocytes. Specifically, this study will look at (1) the mRNA and protein expression profiles, during differentiation, of the various HNF-4 isoforms as well as other factors known to modulate HNF4 activity through Northern, RT-PCR and Western analysis. Likewise, (2) the DNA binding activities of these transcription factors will be assessed through mobility shift and reporter gene assays. Additionally, (3) the functionality of these factors in promoting and/or inhibiting differentiation will be assessed via transfection assays utilizing reporter genes, antisense oligonucleotides, plasmids over- expressing wild type or mutant factors. Lastly, we will investigate specific matrix components and signaling cascades responsible for matrix-induced differentiation. The information derived from the proposed studies will heighten our understanding of the role of extracellular matrix in regulation of cellular differentiation involved in development, regeneration, and cancer.

细胞命运以及其分化状态和表型高度取决于细胞外基质的信号。特别是肝脏和肝细胞培养物，代表了研究细胞生长和分化的适当且方便的模型系统，因为已经确定了几种信号通路和富含肝脏的转录因子，这对于引发和维持肝脏差异化和发展至关重要。 Hepatic growth and differentiation is mediated primarily by the combinatorial actions of liver-enriched transcription factors, foremost of which is HNF-4.. To more clearly understand the molecular mechanisms by which extracellular matrix proteins regulate growth, regeneration and differentiation of hepatocytes, this study aims to investigate the matrixspecific alterations in HNF-4 expression, DNA binding, and transcriptional activity在区分肝细胞中。具体而言，这项研究将研究（1）在分化过程中，各种HNF-4同工型的mRNA和蛋白质表达谱以及已知通过Northern，RT-PCR和Western分析调节HNF4活性的其他因素。同样，（2）这些转录因子的DNA结合活性将通过迁移率转移和报告基因测定来评估。此外，（3）这些因素在促进和/或抑制分化方面的功能将通过使用报告基因，反义寡核苷酸，过度表达野生型或突变因素的质粒，反义寡核苷酸，质粒来评估。最后，我们将研究负责基质引起的分化的特定基质组件和信号级联。源自提出的研究得出的信息将加深我们对细胞外基质在调节发育，再生和癌症中涉及细胞分化的作用的理解。