Enhancer activation mechanisms in cranial neural crest osteoblast differentiation

颅神经嵴成骨细胞分化的增强子激活机制

基本信息

批准号：
10366793
负责人：
Karl Bryan Shpargel
金额：
$ 36.37万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10366793
关键词：
Acetylesterase Anterior Binding Binding Proteins Biochemical Biochemical Genetics Biology Bromodomain Cartilage Cell Count Cell Differentiation process Cell Lineage Cell physiology Cells Cephalic Chromatin Chromatin Remodeling Factor Chromatin Structure Cleft Palate Complex Data Defect Development Disease Downstream Enhancer Ear Embryo Enhancers Enzymes Epigenetic Process Etiology Event Exhibits Face Gene Expression Regulation Genetic Genetic Diseases Genetic Transcription Genome Genomic approach Genomics Histone Acetylation Histone H3 Histones Homologous Gene Human Impairment Kabuki Make-Up Syndrome Knock-in Knock-out Lead Location Lysine Mandible Maps Methylation Methyltransferase Modeling Molecular Molecular Analysis Multipotent Stem Cells Mus Mutate Mutation Neural Crest Neural Crest Cell Neurophysiology - biologic function Osteoblasts Osteogenesis Pathogenesis Pathogenicity Pathway interactions Phenocopy Phenotype Play Point Mutation Post-Translational Protein Processing Process Proteins Proteomics Publishing Reader Regulation Regulatory Pathway Reporter Research Role Shapes Sorting - Cell Movement Therapeutic Tissues Transcriptional Activation Y Chromosome base chromatin remodeling craniofacial craniofacial development craniofacial disorder experience experimental study face bone structure histone methylation histone methyltransferase histone modification improved in vivo loss of function morphogens mutant novel osteoblast differentiation programs protein complex recruit skull base stem cell biology stem cells transcription factor

项目摘要

Project Summary Enhancers play a vital role in initiating and maintaining expression of cell specific transcriptional programs. A key feature in enhancer activation and gene regulation is changing the local chromatin state to facilitate binding of transcriptional co-regulators to enhancers. Chromatin-modifying enzymes that are associated with enhancer function are commonly mutated in human craniofacial disorders including histone methylases, demethylases, acetylases, chromatin readers, and chromatin remodeling proteins. Many of these chromatin-modifying factors biochemically associate and may co-regulate enhancer and transcriptional activation events within neural crest, stem cells from which anterior facial bone and cartilage are derived. We have modeled several of these human craniofacial disorders through tissue specific knockout of chromatin-modifying factors in mouse neural crest cells. The experiments outlined in this proposal will characterize de novo enhancer activation by chromatin profiling of primary cranial neural crest cells undergoing osteoblast differentiation. Neural crest cells deficient in chromatin-modifying machinery will be analyzed molecularly for disruption of enhancer chromatin states and to identify cooperative functions between these factors. This proposal will utilize a combination of biochemical, genetic, and low cell number genomic approaches to elucidate the role of chromatin-based enhancer activation in craniofacial disorder pathogenesis.

项目概要增强子在启动和维持细胞特异性转录程序的表达中发挥着至关重要的作用。一个增强子激活和基因调控的关键特征是改变局部染色质状态以促进结合转录共调节子到增强子。与增强子相关的染色质修饰酶人类颅面疾病中的功能通常发生突变，包括组蛋白甲基化酶、去甲基化酶、乙酰酶、染色质读取器和染色质重塑蛋白。许多染色质修饰因子生物化学上关联并可能共同调节神经内的增强子和转录激活事件嵴，干细胞，源自前面部骨和软骨。我们已经对其中的几个进行了建模通过组织特异性敲除小鼠神经染色质修饰因子来治疗人类颅面疾病嵴细胞。本提案中概述的实验将通过以下方式表征从头增强子激活：进行成骨细胞分化的原代颅神经嵴细胞的染色质分析。神经嵴细胞染色质修饰机制缺陷将进行分子分析，以发现增强子染色质的破坏状态并确定这些因素之间的合作功能。该提案将结合使用生物化学、遗传和低细胞数基因组方法来阐明基于染色质的作用颅面疾病发病机制中的增强子激活。