Mechanisms patterning the midface

中面部图案化机制

• 批准号: 10812765
• 负责人: James Tucker Nichols
• 金额: $ 42.86万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-18 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10812765
• 关键词: 3-Dimensional; Affect; Antibodies; Binding; Binding Sites; Bone structure; Cell Differentiation process; Cells; ChIP-seq; Code; Color; Congenital Abnormality; Data; Development; Disease model; Dysmorphology; Elements; Embryonic Development; Enhancers; Etiology; Eye; Face; Frontonasal dysplasia; Gene Combinations; Gene Expression; Genes; Genetic; Genetic Diseases; Genetic Transcription; Genome; Genomics; Head; Health; Hi-C; Human; Human Genetics; In Situ Hybridization; Jaw; Maps; Mediating; Methods; Mission; Modeling; Molecular; Mutation; National Institute of Dental and Craniofacial Research; Neural Crest Cell; Nose; Orthologous Gene; Pattern; Phenotype; Positioning Attribute; Proteins; Publishing; Reporter; Research; Resolution; Shapes; Skeleton; Specific qualifier value; Structure; System; Testing; Time; Transgenes; Work; Zebrafish; cartilaginous; chromatin immunoprecipitation; craniofacial; craniofacial development; domain mapping; experimental study; gene function; human disease; improved; in vivo; innovation; mutant; paralogous gene; progenitor; promoter; single-cell RNA sequencing; skeletal; success; transcription factor; vertebrate genome; zebrafish development

PROJECT SUMMARY/ABSTRACT: The long-term objective of this proposal is to understand formation of the midface skeleton, both during normal development, and in human genetic disease conditions. The midface consists of the structures around the nose, the eye, and the upper jaw. Human genetic disorders, like frontonasal dysplasia, affect these midface structures. Specifically, the ALX transcription factor encoding genes have been implicated in multiple types of frontonasal dysplasia. Uniquely, we model these diseases in zebrafish by using genetic mutants. We propose that the ALX genes function to specify an identity code which patterns the vertebrate midface. Our model draws from examples like the DLX and HOX codes, which specify identity in the dorsoventral and anteroposterior axes of the craniofacial skeleton, respectively. We propose three specific aims to test our innovative “alx-bullseye code” hypothesis, that nested alx gene expression directly regulates frontonasal skeletal identity. In Aim 1 we define the skeletal structures arising from the alx-bullseye code in wild types using in situ hybridization, live cell tracking, and lineage tracing. Aim 2 will determine whether alx gene combinations function to specify frontonasal identity. To identify these functions, we will compare wild types to alx mutants examining gene expression, skeletal cell differentiation, and misexpression phenotypes. Aim 3 will uncover how alx genes molecularly control cellular identity. We will examine an in vivo Alx direct transcriptional reporter, ChIP-seq to identify Alx protein occupancy across the genome, and Hi-C to reveal enhancer-promoter contacts mediated by Alx. Significance of this proposal is high, the genes that we propose function in the alx bullseye code have direct human orthologs that are associated with midface dysmorphologies. This proposal is innovative, as there has not yet been a patterning code proposed for the vertebrate midface. Success of this proposal will enhance our understanding of midface craniofacial development and human disease. By using the strengths of the zebrafish system to elucidate the molecular, cellular, and genetic mechanisms underlying frontonasal dysplasia, we are fulfilling the mission of the NIDCR to improve human health through research.

项目摘要/摘要：该提案的长期目标是了解 中面部骨骼，无论是在正常发育期间还是在人类遗传疾病条件下。中面部 由鼻子、眼睛和上颌周围的结构组成。人类遗传疾病，例如 额鼻发育不良，影响这些中面部结构。具体来说，ALX转录因子编码基因 与多种类型的额鼻发育不良有关。独特的是，我们对这些疾病进行建模 斑马鱼通过使用基因突变体。我们建议 ALX 基因的功能是指定一个身份代码， 形成脊椎动物中面部的图案。我们的模型借鉴了 DLX 和 HOX 代码等示例，它们指定 分别在颅面骨骼的背腹轴和前后轴上的同一性。我们建议 测试我们创新的“alx-牛眼代码”假设的三个具体目标，即嵌套的 alx 基因表达 直接调节额鼻骨骼特征。在目标 1 中，我们定义了由 使用原位杂交、活细胞追踪和谱系追踪在野生型中进行 alx-bullseye 编码。目标2将 确定alx基因组合是否具有指定额鼻特征的功能。为了识别这些功能， 我们将比较野生型和 alx 突变体，检查基因表达、骨骼细胞分化和 错误表达表型。目标 3 将揭示 alx 基因如何在分子上控制细胞身份。我们将 检查体内 Alx 直接转录报告基因 ChIP-seq，以确定 Alx 蛋白在整个细胞中的占据情况 基因组和 Hi-C 揭示 Alx 介导的增强子-启动子接触。该提案的意义重大 我们建议在 alx bullseye 代码中发挥作用的基因具有相关的直接人类直向同源物 伴有中面部畸形。这个提议是创新的，因为还没有模式代码 建议用于脊椎动物的中面部。该提案的成功将增强我们对 midface 的理解 颅面发育与人类疾病。通过利用斑马鱼系统的优势来阐明 额鼻发育不良的分子、细胞和遗传机制，我们正在履行的使命 NIDCR 通过研究改善人类健康。