Genetic and epigenetic regulation of cranial neural crest differentiation

颅神经嵴分化的遗传和表观遗传调控

• 批准号: 10442617
• 负责人: Kristin Artinger
• 金额: $ 42.6万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-02-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10442617
• 关键词: ATAC-seq; Abnormal Cell; Affect; Binding; Binding Sites; Biological Models; Biology; Cartilage; Cell Differentiation process; Cell Lineage; Cells; Cephalic; Chondrocytes; Chromatin; Chromatin Remodeling Factor; Cleft Lip; Cleft Palate; Competence; Congenital Abnormality; Craniofacial Abnormalities; DNA; DNA Binding; Data; Defect; Development; Developmental Biology; Disease; Due Process; EVI1 gene; Embryo; Enhancers; Epigenetic Process; Etiology; Exhibits; Family; Foundations; GATA1 gene; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Transcription; Genome; Genomics; Genotype; Goals; Growth; Histones; Human; Individual; Knock-out; Knowledge; Limes; Link; Mission; Modification; Mus; Mutation; Nature; Neural Crest; Neural Crest Cell; Palate; Pathway interactions; Penetrance; Phenotype; Play; Process; Protein Family; Proteins; Public Health; Regulation; Research; Role; Signal Transduction; Site; Specific qualifier value; Structural Congenital Anomalies; Syndrome; Testing; Therapeutic; Time; Tissues; United States National Institutes of Health; WNT Signaling Pathway; Zebrafish; bone; cell type; chromatin modification; craniofacial; craniofacial complex; craniofacial structure; design; disability; epigenetic regulation; gain of function; gene network; gene regulatory network; histone methyltransferase; human disease; insight; mutant; neuromechanism; novel; paralogous gene; stem cells; transcription factor; transcriptome; transcriptome sequencing

How cells become specified and differentiate at the correct time and place is a fundamental question in developmental biology. Cranial neural crest cells (cNCCs) are an excellent model system to understand this process due to the multipotent nature of the progenitor cells, generally unrestricted developmental potential with known lineage and derivatives, and defined gene regulatory networks. In addition to the gene networks, epigenetic regulators can affect the expression of numerous target genes and may help to explain the differences in penetrance and phenotype between individuals with the same genotype. This is important since defects in neural crest development underlie many human congenital birth defects, such as cleft lip with or without palate and many craniofacial syndromes. Thus, understanding the genetic and epigenetic regulators in cNCC development is key to understanding how cell fate is determined. We hypothesize that PRDM paralogs regulate global gene expression by regulating downstream targets oppositely, including Wnt pathway components, to control the timing of cartilage/bone differentiation within the cNCC lineage. The rationale for the proposed studies is that an in depth understanding of normal cNCC development will provide insights into normal biology and the etiology of neural crest-associated birth defects, many of which are thought to arise from cNCC abnormalities. We will test this hypothesis in the following specific aims: 1) Test the hypothesis that PRDM proteins act upstream of Wnt signaling to control the timing of cNCC differentiation into chondrocytes. We will test the hypothesis PROM paralog activity is required in cNCCs cell autonomously upstream of Wnt signaling to promote differentiation of chondrocytes. 2) Test the hypothesis that Prdm3 and Prdm16 genetically interact to regulate cNCC gene expression and chromatin accessibility. In Aim 2, hypothesis that Prdm3 and Prdm16 genetically interact to control gene expression via regulating transcription and chromatin modification specifically at cNCC and Wnt gene targets. 3) Test the hypothesis that Prdm3 regulates global gene expression by controlling the timing of genomic accessibility of Prdm16. In Aim 3, we will test the hypothesis that loss of Prdm3 leads to global alterations in chromatin state at cNCC progenitor genes via changes in binding of Prdm16 throughout the genome, which controls the liming of cNCC differentiation into chondrocytes. Together, these studies will reveal basic information of how cNCCs differentiate into specific cell types during development. The results of this proposal have the potential to reveal important new insights into cNCC development and how these processes go wrong in disease, with the hope of providing a foundation for the design of therapeutic strategies for neural crest associated birth defects.

细胞如何在正确的时间和地点变得特化和分化是细胞学中的一个基本问题。 发育生物学。颅神经嵴细胞（cNCC）是理解这一点的优秀模型系统 由于祖细胞的多能性质，通常发育潜力不受限制 具有已知的谱系和衍生物，以及明确的基因调控网络。除了基因网络之外， 表观遗传调节因子可以影响许多靶基因的表达，并可能有助于解释 具有相同基因型的个体之间的外显率和表型差异。这很重要，因为 神经嵴发育缺陷是许多人类先天性缺陷的根源，例如唇裂或 无腭和许多颅面综合征。因此，了解遗传和表观遗传调节因子 cNCC 的发展是理解细胞命运如何决定的关键。我们假设 PRDM 旁系同源物通过相反地调节下游靶标来调节全局基因表达，包括 Wnt 途径成分，以控制 cNCC 谱系内软骨/骨分化的时间。 拟议研究的理由是，深入了解正常的 cNCC 发展将有助于 提供对正常生物学和神经嵴相关出生缺陷的病因学的见解，其中许多是 被认为是由 cNCC 异常引起的。我们将在以下具体目标中检验这个假设：1）检验 PRDM 蛋白作用于 Wnt 信号上游以控制 cNCC 时间的假设 分化成软骨细胞。我们将测试 cNCC 中需要 PROM 旁系同源活动的假设 细胞自主地发挥 Wnt 信号上游的作用，促进软骨细胞的分化。 2) 测试 假设 Prdm3 和 Prdm16 基因相互作用来调节 cNCC 基因表达和 染色质可及性。在目标 2 中，假设 Prdm3 和 Prdm16 基因相互作用以控制基因 通过专门针对 cNCC 和 Wnt 基因靶标的转录和染色质修饰来调节表达。 3) 检验Prdm3通过控制时间来调节全局基因表达的假设 Prdm16 的基因组可及性。在目标 3 中，我们将检验 Prdm3 丢失导致全局 通过整个过程中 Prdm16 结合的变化来改变 cNCC 祖基因的染色质状态 基因组，控制 cNCC 分化为软骨细胞的过程。这些研究共同将 揭示 cNCC 在发育过程中如何分化为特定细胞类型的基本信息。结果 该提案有可能揭示有关 CNCC 发展的重要新见解以及这些 疾病过程出错，希望为治疗策略的设计提供基础 用于神经嵴相关的出生缺陷。