Function of chromatin modifiers in cranial neural crest development

染色质修饰剂在颅神经嵴发育中的作用

基本信息

批准号：
8913662
负责人：
Kristin Artinger
金额：
$ 50.36万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8913662
关键词：
Acetylation Acetyltransferase Affect Behavior Biological Models Cartilage Cell Death Cell Proliferation Cell physiology Cells Centers for Disease Control and Prevention (U.S.)Cephalic Chromatin Cleft Palate Congenital Abnormality Coupled Craniofacial Abnormalities Data Data Set Defect Destinations Development Disease EVI1 gene Elements Enzymes Epigenetic Process Equilibrium Family Family member Ganglia Gene Expression Gene Targeting Genes Genetic Genetic Transcription Genome Genotype Goals Histone H3 Histone-Lysine N-Methyltransferase Histones Human Human Genome In Situ Hybridization In Vitro Kabuki Make-Up Syndrome Knowledge Lead Life Link Live Birth Lysine MLL2 gene Methylation Methyltransferase Mission Modeling Molecular Mouse Strains Mus Mutant Strains Mice Mutation Neural Crest Pathogenesis Pathway Analysis Patients Penetrance Phenotype Population Protein Methyltransferases Proteins Public Health RNA Regulation Regulator Genes Repression Research Role Severities Simpson-Golabi-Behmel syndrome Skeleton Stem cells Syndrome Testing Tissues Transcription Coactivator Transcription Repressor/Corepressor Transcriptional Activation Translating Transplantation United States National Institutes of Health Variant Zebrafish base cartilage development cell motility cellular imaging chromatin remodeling cleft lip and palate craniofacial craniofacial development design disability epigenetic regulation epithelial to mesenchymal transition gene repression genome wide association study histone acetyltransferase histone methyltransferase in vivo innovation insight migration mutant novel novel therapeutics protein function public health relevance skeletal tool

项目摘要

DESCRIPTION (provided by applicant): There is a fundamental gap in our understanding of how defects in chromatin remodeling proteins, methyltransferases and acetyltransferases are causative for human craniofacial phenotypes. This represents an important problem, because craniofacial defects occur frequently in the human population, 1 in every 1000 live births annually in the U.S. (CDC, 2011) and many are associated with epigenetic regulators of the genome. Our long-term goal is to better understand the function of chromatin remodelers during cranial neural crest (cNCC) development. The objective of this application is to determine the mechanism by which two families of epigenetic regulators, KAT2a lysine acetyltransferase and PRDM lysine methyltransferases that regulate each other and act to modify the same H3K9 residue on histone 3, function in zebrafish and mouse cNCC development. We will use two excellent developmental model systems and combine genetic tools with live cell imaging of zebrafish and mouse cNCC behaviors and transcriptional studies to tackle the question of why mutations in Kat2a and Prdms lead to craniofacial abnormalities. The overall hypothesis is that these chromatin modifying enzymes act as opposing transcriptional regulators and function cell autonomously to regulate cNCC proliferation and migration. The rationale for this research is that understanding the mechanism of how KAT2a and PRDMs regulate cNCC development will have the potential to translate into a better understanding of the pathogenesis of craniofacial defects due to mutations in epigenetic regulators, including cleft lip and palate and various syndromes such as Kabuki and SBBYSS that affect the human population. From strong preliminary data, we have designed 3 specific aims: 1) Determine the developmental function of KAT2A and PRDMs in cranial neural crest development, 2) Examine the genetic interaction and regulation of gene targets by KAT2A and PRDMs, and 3) Determine the enzymatic regulation and chromatin state of KAT2A and PRDMs target genes. Under the first aim, we have determined that Prdm1, Prdm3, Prdm16 and Kat2a have craniofacial defects in both mouse and zebrafish. We have the tools and expertise to determine the specific craniofacial defects and to define abnormalities in proliferation and migration of cNCCs. For the second aim, we have generated and obtained most of the zebrafish and mouse strains, and performed transcriptional profiling in both zebrafish and mouse, demonstrating feasibility. For aim three, we have shown analysis of acetylation and methylation states in both tissue and biochemically. Our approach is conceptually innovative in testing a novel hypothesis and technically innovative in the use of live cell imaging and the interplay between two species that model human craniofacial development. The proposed research is significant because it is expected to advance an understanding of how cNCCs form the craniofacial skeleton, which has the potential to inform the treatment of neural crest associated birth defects and craniofacial syndromes.

描述（由适用提供）：我们对染色质重塑蛋白，甲基转移酶和乙酰基转移酶的缺陷的理解存在基本差距。这代表了一个重要的问题，因为颅面缺陷经常发生在人口中，每年在美国每1000个活产中有1个（CDC，2011年），并且许多与基因组的表观遗传调节剂有关。我们的长期目标是更好地了解颅神经rest（CNCC）发育过程中染色质改造的功能。该应用的目的是确定两个表观遗传调节剂的家族Kat2a赖氨酸乙酰基转移酶和PRDM赖氨酸甲基转移酶，它们相互调节并作用于在Zebrafish和House CNCC开发中的组蛋白3，在组蛋白3上进行相同的H3K9住所。我们将使用两个出色的发展模型系统，并将遗传工具与斑马鱼和小鼠CNCC行为的现场细胞成像以及转录研究一起解决，以解决为什么KAT2A和PRDMS突变导致颅面异常的问题。总体假设是，这些染色质修饰酶起作用作为相对的转录调节剂和功能细胞自动调节CNCC的增殖和迁移。这项研究的理由是，了解KAT2A和PRDMS如何调节CNCC开发的机制将有可能使人们可以更好地理解由于表观遗传调节剂的突变，包括裂解唇裂和帕拉特和各种综合症，例如影响Kabuki和Sbbbyss，这些表观遗传调节剂引起的颅面缺陷的发病机理。从强大的初步数据中，我们设计了3个特定目的：1）确定Kat2a和PRDMS在颅神经crest发育中的发育功能，2）检查Kat2a和pRDMS对基因靶标的遗传相互作用和调节，以及3）确定Kat2a和Prdms靶基因的蛋白质调节和染色质状态。在第一个目标下，我们确定PRDM1，PRDM3，PRDM16和KAT2A在小鼠和斑马鱼中均具有颅面缺陷。我们拥有确定特定颅面缺陷并定义CNCC增殖和迁移异常的工具和专业知识。为了第二个目标，我们已经生成并获得了大多数斑马鱼和小鼠菌株，并在斑马鱼和小鼠菌株中进行了转录分析，证明了可行性。对于目标三，我们已经显示了组织和生化的乙酰化和甲基化态的分析。我们的方法在测试新颖的假设和在技术上的使用方面具有创新性细胞成像和模拟人类颅面发育的两个物种之间的相互作用。拟议的研究之所以重要，是因为预计它将促进对CNCC如何形成颅面骨骼的理解，该骨骼有可能告知神经rest相关的先天缺陷和颅面综合征的治疗。