Characterization of the roles and regulation of Draxin in cranial neural crest

Draxin 在颅神经嵴中的作用和调节的表征

• 批准号: 10400365
• 负责人: Erica Hutchins
• 金额: $ 8.56万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 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 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10400365
• 关键词: 3' Untranslated Regions; Affect; Award; COVID-19; Cartilage; Cephalic; Cleft Palate; Craniofacial Abnormalities; Defect; Development; DiGeorge Syndrome; Disease; Down-Regulation; Embryo; Epithelial; Equilibrium; Face; Faculty; Fluorescence Resonance Energy Transfer; Funding; Genetic Transcription; Goals; Head; Knowledge; Lead; Mandibulofacial Dysostosis; Mediating; Mesenchymal; Microscopy; Molecular; Mutation; Names; Neural Crest; Neural Crest Cell; Neural tube; Neuraxis; Parents; Phase; Population; Positioning Attribute; Proteins; Publications; Publishing; RNA immunoprecipitation sequencing; RNA-Binding Proteins; Regulation; Research; Role; Secure; Signal Transduction; Skeleton; Techniques; Time; Translational Research; Universities; WNT Signaling Pathway; Work; career; craniofacial; craniofacial structure; epithelial to mesenchymal transition; experimental study; in vivo; malformation; migration; molecular imaging; neuroepithelium; post-doctoral training; programs; public health relevance; single molecule; skills; spatiotemporal; stem cell population; tenure track

PROJECT SUMMARY/ABSTRACT The neural crest (NC) is a stem cell population that originates within the forming central nervous system. NC cells delaminate from the neuroepithelium by undergoing a spatiotemporally regulated epithelial— mesenchymal transition (EMT) to exit from the neural tube. Cranial NC cells, which arise in the head region of the embryo and are the only NC population in vivo with the ability to differentiate into craniofacial skeleton and cartilage, are indispensable for the development of the face; mutations affecting NC development result in numerous diseases and malformations affecting the craniofacial structures. The focus of my postdoctoral work has been to study the mechanisms that control and facilitate cranial NC EMT. During the first phase of my postdoctoral training, I have shown that this developmental EMT program is controlled by temporally restricted expression of the Wnt antagonist, Draxin. A hallmark of Draxin's function during EMT is its transient expression and rapid downregulation; perdurance of Draxin has deleterious effects on cranial NC EMT through dysregulation of downstream targets of canonical Wnt signaling. Through the support of the K99, I discovered that the transience of Draxin expression in cranial NC is mediated post-transcriptionally via its 3'-untranslated region (UTR). Importantly, Draxin is stabilized by the RNA-binding protein Elavl1/HuR at the premigratory stage, then targeted to cytoplasmic processing bodies (P-bodies) for decay to drive proper cranial NC EMT. Collectively, these discoveries begin to unravel a new mechanism whereby cranial NC EMT is regulated through post-transcriptional regulatory mechanisms balancing stability and decay of a molecular rheostat, Draxin. COVID-19 research restrictions and university closures severely delayed my career plans and development. Through the support of the K99, I completed many of the goals proposed in Aims 1 and 3 of my original proposal, which sought to illuminate the interaction between Draxin and Wnt signaling, and the regulation Draxin expression, respectively. However, COVID-19 research restrictions severely delayed the completion of Aim 1 and progress of Aim 2, which sought to apply time-lapse and advanced microscopy techniques (e.g. FRET) to more fully explore Draxin function. Further, completion of Aim 3 and publication of these studies requires additional experiments in single-molecule imaging and RIP-seq. A funding extension would allow me to develop critical new skills in advanced microscopy and RIP-seq to gain a mechanistic understanding of Draxin activity during cranial NC EMT, and allow me to comple the revision experiments necessary to publish the work performed under the parent K99 award to help me secure a tenure-track faculty position, establish a vibrant independent research program in the developmental signaling field, and better equip me with the knowledge necessary to transition into the study of cranial NC development and migration.

项目摘要/摘要 神经rest（NC）是一种干细胞种群，起源于中枢神经内 系统。 NC细胞通过进行空间调节的上皮 - 从神经上皮分层 - 间充质转变（EMT）从神经管中退出。颅NC细胞，在 胚胎是体内唯一的NC种群，能够分化为颅面骨骼和 软骨，对于面部的发展是必不可少的；影响NC发展的突变导致 影响颅面结构的许多疾病和畸形。我的博士后工作的重点 已经研究了控制和促进颅NC EMT的机制。在我的第一阶段 博士后培训，我已经证明了此开发EMT程序受临时限制的控制 Wnt拮抗剂Draxin的表达。 Draxin在EMT期间功能的标志是其瞬态表达式 和快速下调； Draxin的Perdurance通过 规范Wnt信号传导下游目标的失调。在K99的支持下，我发现了 Draxin在颅NC中表达的瞬态是通过其3'-非翻译后的后介导的 区域（UTR）。重要的是，德拉克斯在前移民的RNA结合蛋白Elavl1/hur稳定 阶段，然后靶向细胞质加工体（p-bodies），以驱动适当的颅NC EMT。 总的来说，这些发现开始揭示一种新机制，从而调节了颅内NC EMT 通过转录后调节机制平衡稳定性和分子变势的衰减， 德拉辛。 Covid-19的研究限制和大学关闭严重延迟了我的职业计划， 发展。通过K99的支持，我完成了目标1和3中提出的许多目标 原始提议，阐明了Draxin和Wnt信号之间的相互作用，以及 调节Draxin表达。但是，COVID-19的研究限制严重延迟了 AIM 1的完成和AIM 2的进展，这是应用延时和高级显微镜的感觉 技术（例如FRET）以更充分的探索Draxin功能。此外，AIM 3的完成和发布 这些研究需要在单分子成像和RIP-seq中进行其他实验。资金扩展 我可以在高级显微镜和RIP-seq方面发展重要的新技能，从而获得机理 了解颅NC EMT期间的Draxin活性，并让我完成修订实验 出版根据家长K99奖所做的工作有必要的工作，以帮助我确保终身教师 位置，在发育信号领域建立充满活力的独立研究计划，并更好 为我提供过渡到颅NC发展和移民研究所需的知识。