CDKN2B as a Novel Epigenetically Regulated Gene in Idiopathic Pulmonary Fibrosis

CDKN2B 作为特发性肺纤维化中的新型表观遗传调控基因

• 批准号: 9247799
• 负责人: STEVEN K HUANG
• 金额: $ 37.8万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9247799
• 关键词: Address; Alpha Cell; Apoptosis; Architecture; Biological; Biology; CDKN1A gene; Cardiovascular Diseases; Cell Cycle; Cell Cycle Progression; Cell Proliferation; Cells; Cicatrix; Cyclin-Dependent Kinase Inhibitor; DNA; DNA Methylation; Data; Development; Disease; Disease Pathway; Effector Cell; Epigenetic Process; Etiology; Exhibits; Extracellular Matrix; Family; Family member; Fibroblasts; Fibrosis; Future; Gene Expression; Gene Targeting; Genes; Goals; Grant; Hamman-Rich syndrome; Hypermethylation; In Vitro; Individual; Investigation; Lung; Malignant Neoplasms; Methylation; Mission; Modification; Myofibroblast; Nature; Normal tissue morphology; Pathogenesis; Pathology; Pathway interactions; Patients; Phenotype; Play; Production; Progressive Disease; Public Health; Pulmonary Fibrosis; Research; Research Personnel; Resistance; Role; Surveys; Testing; Tumor Suppressor Genes; United States National Institutes of Health; Untranslated RNA; base; cell type; epigenetic regulation; epigenomics; experimental study; genome wide association study; genome wide methylation; improved; inhibitor/antagonist; innovation; insight; knowledge base; methylation pattern; mouse model; novel; public health relevance; treatment strategy; tumor; Address; Alpha Cell; Apoptosis; Architecture; Biological; Biology; CDKN1A gene; Cardiovascular Diseases; Cell Cycle; Cell Cycle Progression; Cell Proliferation; Cells; Cicatrix; Cyclin-Dependent Kinase Inhibitor; DNA; DNA Methylation; Data; Development; Disease; Disease Pathway; Effector Cell; Epigenetic Process; Etiology; Exhibits; Extracellular Matrix; Family; Family member; Fibroblasts; Fibrosis; Future; Gene Expression; Gene Targeting; Genes; Goals; Grant; Hamman-Rich syndrome; Hypermethylation; In Vitro; Individual; Investigation; Lung; Malignant Neoplasms; Methylation; Mission; Modification; Myofibroblast; Nature; Normal tissue morphology; Pathogenesis; Pathology; Pathway interactions; Patients; Phenotype; Play; Production; Progressive Disease; Public Health; Pulmonary Fibrosis; Research; Research Personnel; Resistance; Role; Surveys; Testing; Tumor Suppressor Genes; United States National Institutes of Health; Untranslated RNA; base; cell type; epigenetic regulation; epigenomics; experimental study; genome wide association study; genome wide methylation; improved; inhibitor/antagonist; innovation; insight; knowledge base; methylation pattern; mouse model; novel; public health relevance; treatment strategy; tumor

 DESCRIPTION (provided by applicant): Although the etiology of idiopathic pulmonary fibrosis (IPF) is unknown and may involve diverse mechanisms, excessive fibroproliferation remains a central hallmark in IPF pathology. Among the many cell types that play critical roles in disease pathogenesis, fibroblasts are ultimately the effector cell of fibrosis. Fibroblasts from IPF patiens have been shown by multiple investigators to exhibit a profibrotic in vitro phenotype; these observations were often made even when cells were cultured after multiple cell passages, suggesting that epigenetic modifications may be responsible for these phenotypic changes. Through a global survey of DNA methylation patterns in IPF fibroblasts, CDKN2B was discovered to be one of the most differentially methylated genes in IPF fibroblasts. Although never previously investigated in the context of IPF, CDKN2B has been shown to be highly differentially methylated in many cancers and to regulate cell proliferation and cell cycle progression in tumors. Our objectives are to determine the mechanism by which hypermethylation and diminished CDKN2B expression contributes to the activated phenotype of IPF cells, and to understand how its expression is regulated by DNA methylation, and how methylation may be dictated by the expression and actions of a long noncoding RNA (lncRNA). Our central hypothesis is that hypermethylation of CDKN2B, driven by the lncRNA CDKN2B-antisense (AS) 1, contributes to the diminished expression of CDKN2B in IPF fibroblasts, which leads to increased myofibroblast differentiation and apoptosis resistance. This project has three specific aims: 1) Determine how diminished expression of CDKN2B contributes to a profibrotic phenotype in IPF fibroblasts; 2) Determine how DNA methylation modulates the expression of CDKN2B; and 3) Elucidate the mechanisms by which CDKN2B-AS1 contributes to the DNA hypermethylation of CDKN2B in IPF fibroblasts. The biological actions of CDKN2B in pulmonary fibrosis will be explored by examining primary fibroblasts from normal lung and patients with IPF and by mouse modeling experiments. DNA methylation analysis will be performed, and the role of lncRNAs in regulating the DNA methylation of CDKN2B will be examined. This project is innovative because it seeks to understand the function, and additionally the epigenetic regulation, of a gene that may be critical to fibroproliferation, but has never been previously investigated in IPF. Completion of these studies is expected to establish the biological importance of CDKN2B to IPF fibroblast biology, and elucidate mechanisms that regulate its expression and contribute to how differential DNA methylation patterns are established. These findings will provide not only a novel gene target for future therapy, but also advance our understanding of how DNA methylation changes arise and contribute to gene expression differences in IPF.

 描述（由适用提供）：尽管特发性肺纤维化（IPF）的病因尚不清楚，并且可能涉及不同的机制，但过量的纤维增殖仍然是IPF病理学的核心标志。在许多在疾病发病机理中起关键作用的细胞类型中，成纤维细胞最终是纤维化的效应细胞。来自IPF patiens的成纤维细胞已被多个研究人员显示出表现出纤维化的体外表型。这些观察通常是即使在多个细胞通道后培养细胞的情况下也经常进行的，这表明表观遗传修饰可能是这些表型变化的原因。通过对IPF成纤维细胞中DNA甲基化模式的全球调查，发现CDKN2B是IPF成纤维细胞中最不同的甲基化基因之一。尽管以前从未在IPF的背景下研究过CDKN2B，但在许多癌症中我们被证明是高度差异化的，并调节了肿瘤中细胞增殖和细胞周期进程。我们的目标是确定高甲基化和CDKN2B表达降低的机制有助于IPF细胞的激活表型，并了解其表达如何受DNA甲基化调节，以及如何通过长的非编码RNA（LNCRNA）的表达和作用来决定甲基化。我们的中心假设是，由LNCRNA CDKN2B-抗态驱动的CDKN2B的高甲基化（AS）1，导致CDKN2B在IPF成纤维细胞中的表达降低，从而导致肌纤维细胞差异分化和抑制性增加。该项目具有三个特定的目的：1）确定CDKN2B表达的降低是如何促进IPF成纤维细胞中纤维化表型的； 2）确定DNA甲基化如何调节CDKN2B的表达； 3）阐明CDKN2B-AS1在IPF成纤维细胞中促进CDKN2B的DNA高甲基化的机制。 CDKN2B在肺纤维化中的生物学作用将通过检查正常肺和IPF患者的原发性成纤维细胞以及小鼠建模实验来探索。将进行DNA甲基化分析，并将检查LNCRNA在确定CDKN2B的DNA甲基化中的作用。该项目具有创新性，因为它试图理解可能对纤维增生至关重要的基因的功能，又是表观遗传调节，但以前从未在IPF中进行过研究。这些研究的完成有望确定CDKN2B对IPF成纤维细胞生物学的生物学重要性，并阐明调节其表达的机制并有助于建立差异DNA甲基化模式。这些发现不仅将为将来的治疗提供一个新的基因靶标，而且还可以提高我们对DNA甲基化变化的理解，并有助于IPF中的基因表达差异。