Molecular mechanisms regulating cranial sensory development

调节颅感觉发育的分子机制

• 批准号: 10679365
• 负责人: Helen Ruth Maunsell
• 金额: $ 4.85万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-14 至 2026-08-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679365
• 关键词: ATAC-seq; Address; Affect; Anterior; Cartilage; Cell Lineage; Cells; Central Nervous System; Cephalic; Characteristics; Chromatin; Classification; Cleft lip with or without cleft palate; Complex; Congenital Abnormality; Data; Development; Disease; Embryo; Embryonic Development; Epidermis; Epigenetic Process; Face; Functional disorder; Genes; Genetic; Genetic Transcription; Goals; Impairment; Jaw; Knockout Mice; Knowledge; Label; Labyrinth; Methods; Molecular; Mus; Mutation; Nerve; Neural Crest; Neural Crest Cell; Neuroglia; Neurons; Null Lymphocytes; Olfactory Epithelium; Organ; Palate; Pathway interactions; Peripheral Nervous System; Phenotype; Population; Positioning Attribute; Process; Reconstructive Surgical Procedures; Reporter; Role; Sensory; Signal Induction; Signal Transduction; Signaling Molecule; Specific qualifier value; Structural defect; Structure; Syndrome; Testing; Time; Tissues; Transgenic Mice; Work; Zebrafish; bone; cell type; craniofacial; craniofacial development; craniofacial disorder; extracellular; face bone structure; gastrulation; genetic manipulation; in vivo; insight; multiple omics; multipotent cell; mutant; neural plate; neurosensory; novel; novel marker; progenitor; programs; response; segregation; single-cell RNA sequencing; stem cells; transcription factor

Project Summary Worldwide, more than a third of congenital birth defects are classified as craniofacial disorders. These disorders present diversely, from abnormal formation of facial features such as the jaw or palate, to impaired sensory function. Despite advances in surgical reconstruction, there is still a poor understanding of the molecular pathophysiology that underlies distinct phenotypes. Four multipotent cell lineages are the precursors to all craniofacial cell types, forming in an anterior region of the embryo known as the neural plate border. The central-most lineages— pre-placodal and neural crest— are especially key to craniofacial development, with pre-placodal cells giving rise to supporting and sensorineural cell types, while neural crest cells become the neurons and glia of the peripheral nervous system and the bones and cartilage of all facial structures. Yet identifying the factors responsible for the initial segregation of pre-placodal and neural crest lineages has been difficult, given that the two populations intermingle closely. This study thus aims to use a genetic lineage- tracing approach to isolate the pre-placodal and crest lineages and investigate the roles of two candidate molecules in pre-placodal specification. Our lab previously discovered that the Foxi3 transcription factor is transiently expressed in border cells and that genetic deletion of Foxi3 primarily affects placode-derived structures, including loss of the inner ear. Preliminary lineage tracing with a Foxi3CreER conditional reporter mouse line generated in the lab revealed that normal Foxi3-expressing border cells mostly become placode derivatives, but some mutant cells take on alternative border lineage fates. From this data, we hypothesize that Foxi3 directly specifies the pre-placodal lineage. We will test this hypothesis by further analyzing the fate of Foxi3 mutant border cells, using lineage tracing with our Foxi3CreER mice. We will also use a single-cell multiomic approach to assess concurrent transcriptional and epigenetic changes in Foxi3 functionally null border cells. Extracellular signaling also influences border lineages: notably, BMP signals are known to induce neural crest cells. Given the similar BMP levels to which crest and pre-placodal cells are exposed, we hypothesize that BMP also affects placode cell specification. We will test this hypothesis by an in vivo, genetic manipulation of BMP targeted to pre-placodal (Foxi3CreER) versus crest (Zic5CreER) border cells. Our work introduces the first method to isolate and study the mammalian pre-placodal lineage and will shed light on critical factors that direct neural plate border cell fate. We will also gain insight to molecular changes that facilitate fate transitions at the border, with the potential to identify genes whose mutation could result in specific kinds of disruption to craniofacial development.

项目摘要 在世界范围内，超过三分之一的先天性出生缺陷被归类为颅面疾病。这些 从面部特征如颌或腭的异常形成，到受损的 感觉功能尽管在外科重建方面取得了进展，但仍然缺乏对 不同表型的分子病理生理学基础。四种多能细胞谱系是前体细胞 所有颅面细胞类型，形成在胚胎的前部区域，称为神经板边界。的 最中心的谱系-前基板和神经嵴-是颅面发育的关键， 前基板细胞产生支持细胞和感觉神经细胞类型，而神经嵴细胞则成为支持细胞和感觉神经细胞类型。 周围神经系统的神经元和神经胶质以及所有面部结构的骨骼和软骨。然而 确定负责前基板和神经嵴谱系的初始分离的因素， 很难，因为这两个群体密切混合。因此，这项研究旨在使用遗传谱系- 追踪方法分离前基板和嵴谱系并研究两个候选者的作用 前基板规格的分子。我们的实验室先前发现Foxi 3转录因子是 在边缘细胞中瞬时表达，并且Foxi 3的遗传缺失主要影响基板来源的 结构，包括内耳的丧失。使用Foxi 3CreER条件性报告基因进行初步谱系追踪 在实验室中产生的小鼠系显示，正常的表达Foxi 3的边缘细胞大多成为基板， 这些突变体细胞具有不同的衍生物，但一些突变体细胞具有替代的边界谱系命运。根据这些数据，我们假设， Foxi 3直接指定前基板谱系。我们将通过进一步分析 使用我们的Foxi 3CreER小鼠的谱系追踪的Foxi 3突变体边缘细胞。我们还将使用单细胞 多组学方法评估Foxi 3功能缺失的同时转录和表观遗传变化 边界细胞细胞外信号也影响边界谱系：值得注意的是，已知BMP信号诱导 神经嵴细胞考虑到嵴和前基板细胞暴露的BMP水平相似， 假设BMP也影响基板细胞特化。我们将通过体内遗传学方法来验证这一假设。 靶向前基板（Foxi 3CreER）与嵴（Zic 5CreER）边缘细胞的BMP的操作。我们的工作 介绍了第一种分离和研究哺乳动物前基板谱系的方法，并将阐明 决定神经板边缘细胞命运的关键因素。我们还将深入了解分子变化， 促进边界的命运转变，有可能确定其突变可能导致 对颅面发育的特殊破坏。