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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（由申请人提供）：我们对染色质重塑蛋白、甲基转移酶和乙酰转移酶的缺陷如何导致人类颅面表型的理解存在根本性差距。这代表了一个重要的问题，因为颅面缺陷在人群中频繁发生，在美国每年每1000个活产婴儿中就有1个（CDC，2011），并且许多与基因组的表观遗传调节因子相关。我们的长期目标是更好地了解颅神经嵴（cNCC）发育过程中染色质重塑的功能。本申请的目的是确定两个家族的表观遗传调节因子KAT2a赖氨酸乙酰转移酶和PRDM赖氨酸甲基转移酶在斑马鱼和小鼠cNCC发育中发挥作用的机制，所述表观遗传调节因子KAT2a赖氨酸乙酰转移酶和PRDM赖氨酸甲基转移酶相互调节并用于修饰组蛋白3上的相同H3K9残基。我们将使用两个优秀的发育模型系统，并将联合收割机遗传工具与斑马鱼和小鼠cNCC行为的活细胞成像和转录研究相结合，以解决Kat2a和Prdms突变导致颅面异常的原因。总的假设是这些染色质修饰酶作为相反的转录调节因子并且自主地发挥细胞功能以调节cNCC增殖和迁移。这项研究的基本原理是，了解KAT2a和PRDMs如何调节cNCC发育的机制将有可能转化为更好地理解由于表观遗传调节因子突变引起的颅面缺陷的发病机制，包括唇腭裂和各种综合征，如影响人类的歌舞伎和SBBYSS。根据强有力的初步数据，我们设计了3个具体目标：1）确定KAT2A和PRDMs在颅神经嵴发育中的发育功能，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靶基因的酶促调节和染色质状态。在第一个目标下，我们已经确定Prdm1，Prdm3，Prdm16和Kat2a在小鼠和斑马鱼中都有颅面缺陷。我们有工具和专业知识来确定特定的颅面缺陷，并确定cNCC增殖和迁移的异常。对于第二个目标，我们已经产生并获得了大多数斑马鱼和小鼠品系，并在斑马鱼和小鼠中进行转录谱分析，证明了可行性。对于第三个目标，我们已经展示了组织和生物化学中乙酰化和甲基化状态的分析。我们的方法在概念上是创新的，在测试一个新的假设和技术上的创新，在使用现场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。这项研究意义重大，因为它有望促进对cNCC如何形成颅面骨骼的理解，这有可能为神经嵴相关出生缺陷和颅面综合征的治疗提供信息。