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 在全球范围内，超过三分之一的先天性先天缺陷被归类为颅面疾病。这些 从异常形成面部特征（例如下颌或味觉）到障碍，到障碍都有多种疾病 感官功能。尽管手术重建方面取得了进步，但对 分子病理生理学，其基础是不同的表型。四个多透明细胞谱系是前体 对于所有颅面细胞类型，形成在称为神经板边界的胚胎的前区域。 中心最多的谱系 - placodal和神经波峰 - 特别是颅面发育的关键 替代前的细胞引起了支持和感觉神经细胞类型，而神经rest细胞成为 周围神经系统的神经元和神经元以及所有面部结构的骨骼和软骨。然而 确定负责初始隔离的因素，是平台前和神经rest谱系的最初分离 鉴于这两个人群密切相互交织，很难。因此，这项研究旨在使用遗传谱系 追踪方法，以隔离抛射线前和波峰谱系并研究两个候选者的作用 平位前规范中的分子。我们的实验室以前发现FOXI3转录因子是 在边界细胞中瞬时表达，FOXI3主要的遗传缺失会影响placode衍生 结构，包括内耳的丧失。使用FOXI3CREER条件记者的初步谱系跟踪 实验室中生成的鼠标线表明，正常表达FOXI3的边框细胞大多成为placode 衍生物，但一些突变细胞采用替代边界谱系命运。从这些数据中，我们假设 FOXI3直接指定了前姿态谱系。我们将通过进一步分析命运来检验这一假设 FOXI3突变体细胞，使用我们的FOXI3CREER小鼠进行谱系跟踪。我们还将使用一个单细胞 评估FOXI3的并发转录和表观遗传变化的多态方法在功能上无效 边界细胞。细胞外信号传导也会影响边界谱系：尤其是，已知BMP信号会影响 神经rest细胞。鉴于to暴露了波峰和平台前细胞的相似BMP水平，我们 假设BMP还会影响Placode单元格的规范。我们将通过体内的通用，一般性的一般性检验该假设 对靶向前平台（FOXI3CREER）与Crest（ZIC5CREER）边界细胞的BMP操纵。我们的工作 介绍了第一种隔离和研究哺乳动物前血统谱系的方法，并将阐明 直接神经板边界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，并有可能识别其突变可能导致的基因 颅面发育的特定类型的破坏。