Epigenetic mechanisms of tumor formation in Beckwith-wiedemann Syndrome

Beckwith-wiedemann 综合征肿瘤形成的表观遗传机制

基本信息

批准号：
9813688
负责人：
NORA I ENGEL
金额：
$ 7.93万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9813688
关键词：
3-Dimensional Address Affect Beckwith-Wiedemann Syndrome Binding Sites Biological Assay CCCTC-binding factor CDKN1C gene CRISPR/Cas technology Cell Cycle Cell Line Cells Chimeric Proteins Chromatin Chromosomes Chronology DNA Methylation DNA Sequence DNA Sequence Alteration Data Development Dimensions Disease Engineering Enhancers Epigenetic Process Event Exhibits Fetal Macrosomia Fibroblasts Future Gene Expression Genes Genetic Transcription Growth Guide RNA Health Human Human Cell Line Individual Ligation Maintenance Malignant Childhood Neoplasm Malignant Neoplasms Maps Medical Methylation Modification Molecular Conformation Mus Mutation Normal Cell Nucleic Acid Regulatory Sequences Online Mendelian Inheritance In Man Patients Phenotype Positioning Attribute Predisposition Publishing Regulatory Element Resolution Risk Role Site Syndrome Technology Testing Therapeutic Time Series Analysis Untranslated RNA carcinogenesis cell growth chromatin immunoprecipitation chromosome conformation capture comparative demethylation design epigenetic therapy epigenomics experience experimental study genome-wide high risk imprint inhibitor/antagonist insight new technology next generation novel profiles in patients promoter targeted treatment tumor

项目摘要

Beckwith-Wiedeman Syndrome (BWS) is a fetal overgrowth disorder which confers high risk for pediatric cancers. Loss of DNA methylation (LOM) in a key regulatory region of the KCNQ1 domain resulting in silencing of a critical cell cycle inhibitor, CDKN1C, occurs in a large group of patients. However, it is unknown how methylation is disrupted, how it affects other chromatin features and how it leads to tumor development. Thus, BWS provides a useful paradigm for addressing epigenetic dysregulation in cancer and offers an opportunity to test novel epigenetic therapeutic strategies. We will perform conformational, epigenetic and expression studies to determine the status of known promoters, enhancers and insulators and to identify novel regulatory sequences in the human KCNQ1 domain. We will also perform comparative analyses between normal fibroblasts and cells from BWS patients exhibiting LOM with co-occurring deletions. These comparisons will elucidate how LOM affects expression of critical genes in carcinogenesis and explain how the sequences spanned by patient deletions affect conformation and DNA methylation. We will engineer similar deletions in normal fibroblasts and epigenetically engineer normal cells to demethylate the DNA sequences affected in BWS. In addition, we will exploit epigenetic fusion proteins to rescue the epigenetic and transcriptional profiles of the patient cells. Because current epigenetic therapies act in a genome-wide manner, successful modification of specific loci with epigenetic editors portends a highly targeted therapy applicable to any cancer with deregulated gene expression.

贝克维斯·妻子综合症（BWS）是一种胎儿过度生长障碍，赋予小儿高风险癌症。在KCNQ1结构域的关键调节区域中DNA甲基化（LOM）的丧失，导致沉默在一大群患者中，关键细胞周期抑制剂CDKN1C发生。但是，这是未知的甲基化被破坏，如何影响其他染色质特征以及如何导致肿瘤发育。因此， BWS为解决癌症的表观遗传失调提供了有用的范例，并提供了测试新型表观遗传治疗策略的机会。我们将执行构象，表观遗传和表达研究以确定已知启动子，增强子和绝缘体的状态，并确定新颖的人类KCNQ1域中的调节序列。我们还将进行比较分析来自BWS患者的正常成纤维细胞和细胞，表现出具有共同缺失的LOM。这些比较将阐明LOM如何影响癌变中关键基因的表达，并解释如何患者缺失跨越的序列会影响构象和DNA甲基化。我们将设计类似正常成纤维细胞和表观遗传学的正常细胞中缺失以将DNA序列脱甲基在BWS中受到影响。此外，我们将利用表观遗传融合蛋白来营救表观遗传和患者细胞的转录曲线。因为当前的表观遗传疗法以全基因组的方式起作用，所以通过表观遗传编辑者成功修改特定基因座，预示着一种适用于任何具有失调基因表达的癌症。