Functions of PRDM Histone Methyltransferases during Cartilage Development in the Craniofacial Skeleton

来源于颅神经脊细胞的软骨细胞长出软骨结构，形成颅面 骷髅。这些细胞必须经历许多细胞过程，包括凝结、定向、 插入、增殖、分化和成熟，然后形成用作支架的模板 用于随后的骨骼形成。基因调控网络(GRN)和信号通路，控制这些 流程需要受到严格监管。软骨细胞过程中GRN或信号模块的任何变化 成熟和分化会损害发育中的头面部组织的骨骼完整性 导致先天性畸形的病因，包括但不限于唇裂伴或不伴腭裂， 下颌骨发育不全，颅缝早闭。我有兴趣了解染色质修饰剂Prdm3是如何 和Prdm16，表观遗传控制颅面发育过程中的空间和时间基因表达。这个 这项提案中概述的目标是利用分子、遗传和表观遗传工具在斑马鱼和老鼠身上进行测试 Prdm3和Prdm16作用于Wn/β-catenin上游以适当平衡软骨细胞的假说 在头面部复合体形成过程中的功能。在目标1中，分子机制控制 Wnt/β-Catenin在斑马鱼头面部发育过程中软骨细胞中的转录活性将被定义。 通过调节Wnt/β-连环蛋白对软骨细胞极性和分化的保守功能将是 在哺乳动物的头面部复合体中被鉴定(目标2)，最后，保守的Prdm3-和 PRDM16调节的脊椎动物颅面间充质中典型的Wnt/β-catenin增强子将 被评估(目标3)。完成这些目标将提供对这些表观遗传修饰因子如何控制的洞察 特定的GRN和信号模块(Wnt/β-catenin)促进正常的软骨形成 头面部骨骼。重要的是，这个项目还将提供这些损失背后的机械性洞察 各种因素导致了颅面部疾病的发展。随着职业生涯的发展制定研究型培训计划 本提案中概述的发展和指导计划旨在为我的职业生涯奠定基础 目标是成为一家顶级研究机构的独立研究员。在K99阶段，我将收到 Kristin Artinger在斑马鱼生物学方面的指导，以及成员对我的建议的指导 小鼠颅面生物学和生物信息学分析委员会。广泛的职业发展计划 与促进赠款撰写、科学交流以及领导和指导技能的活动保持一致 通过MOSAIC计划，将有助于我在R00阶段过渡到独立的教员职位。 科罗拉多大学安舒茨医学院的颅面部系提供了一种特殊的 拥有无数资源的环境来进行这项研究和培训计划，这将指导我的成功 朝着实现我的职业目标迈进。

Chondrocytes derived from cranial neural crest cells give rise to cartilaginous structures that form the craniofacial skeleton. These cells must undergo numerous cellular processes including condensation, orientation, intercalation, proliferation, differentiation, and maturation before forming a template that will serve as a scaffold for subsequent bone formation. The gene regulatory networks (GRNs) and signaling pathways, controlling these processes need to be tightly regulated. Any alteration to the GRNs or signaling modules during chondrocyte maturation and differentiation can compromise the skeletal integrity of the developing craniofacial tissues and contribute to the etiology of congenital defects including but not limited to cleft lip with or without cleft palate, mandibular hypoplasia, craniosynostosis. I am interested in understanding how the chromatin modifiers, Prdm3 and Prdm16, epigenetically control spatial and temporal gene expression during craniofacial development. The aims outlined in this proposal utilize molecular, genetic and epigenetic tools in both zebrafish and mice to test the hypothesis that Prdm3 and Prdm16 act upstream of Wn/β-catenin to properly balance chondrocyte functionality during the formation of the craniofacial complex. In Aim 1, the molecular mechanisms controlling Wnt/β-catenin transcriptional activity in chondrocytes during zebrafish craniofacial development will be defined. The conserved functions of chondrocyte polarity and differentiation through regulation of Wnt/β-catenin will be identified in the mammalian craniofacial complex (Aim 2), and lastly, the functions of conserved Prdm3- and Prdm16-regulated canonical Wnt/β-catenin enhancers across vertebrates in the craniofacial mesenchyme will be assessed (Aim 3). Completion of these aims will provide insight on how these epigenetic modifiers control specific GRNs and signaling modules (Wnt/β-catenin) to facilitate proper chondrogenesis in formation of the craniofacial skeleton. Importantly, this project will also provide mechanistic insight behind how loss of these factors contributes to the development of craniofacial disorders. The research training plan along with the career development and mentorship plan outlined in this proposal are designed to provide the foundation for my career goal of becoming an independent investigator at a top research institution. During the K99 phase, I will receive mentorship in zebrafish biology from Kristin Artinger, as well as guidance from members on my advisory committee for mouse craniofacial biology and bioinformatics analysis. An extensive career development plan with activities promoting grant writing, scientific communication, and leadership and mentoring skills, in alignment with the MOSAIC program, will facilitate my transition to an independent faculty position during the R00 phase. The Craniofacial Department at the University of Colorado Anschutz Medical Campus offers an exceptional environment with countless resources to conduct this research and training plan that will guide my success toward reaching my career goals.