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

PRDM 组蛋白甲基转移酶在颅面骨骼软骨发育过程中的功能

• 批准号: 10551839
• 负责人: Lomeli Carpio Shull
• 金额: $ 6.41万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10551839
• 关键词: ATAC-seq; Advisory Committees; Automobile Driving; BCL9 gene; Binding; Bioinformatics; Biological Assay; Biology; Bone structure; Cartilage; Cell Culture Techniques; Cell Polarity; Cell physiology; Cells; Cephalic; Chondrocytes; Chondrogenesis; Chromatin; Chromatin Remodeling Factor; Cleft Lip; Cleft Palate; Cleft lip with or without cleft palate; Colorado; Communication; Congenital Abnormality; Craniofacial Abnormalities; Craniosynostosis; Data; Data Set; Development; Development Plans; Enhancers; Environment; Epigenetic Process; Equilibrium; Etiology; Face; Faculty; Family; Foundations; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Goals; Grant; Immunoprecipitation; In Vitro; Institution; Leadership; Maintenance; Mandible; Medical; Mentors; Mentorship; Mesenchyme; Micrognathism; Modeling; Molecular; Mus; Neural Crest; Neural Crest Cell; Nuclear; Osteogenesis; Pathway interactions; Phase; Physical condensation; Positioning Attribute; Process; Proliferating; Proteins; Public Health; Regulation; Reporter; Repression; Research; Research Personnel; Research Training; Resources; Role; Signal Pathway; Signal Transduction; Skeleton; Structure; System; Testing; Time; Training; Universities; Vertebrates; Writing; Zebrafish; beta catenin; career; career development; cartilage development; cartilaginous; craniofacial; craniofacial complex; craniofacial development; craniofacial disorder; craniofacial structure; craniofacial tissue; design; differential expression; experimental study; gene regulatory network; histone methyltransferase; in vivo; insight; intercalation; interest; loss of function; member; mutant; overexpression; programs; scaffold; skeletal; skills; success; tool; transcription factor; transcriptome; transcriptome sequencing; transcriptomics

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.

来源于颅神经嵴细胞的软骨细胞产生形成颅面的软骨结构 骷髅这些细胞必须经历许多细胞过程，包括浓缩，定向， 插入、增殖、分化和成熟，然后形成用作支架的模板 用于随后的骨形成。基因调控网络（GRNs）和信号通路，控制这些 需要严格监管这些流程。软骨细胞生长过程中GRNs或信号模块的任何改变 成熟和分化可能损害发育中的颅面组织的骨骼完整性， 有助于先天性缺陷的病因学，包括但不限于唇裂伴或不伴腭裂， 下颌发育不全颅缝早闭我有兴趣了解染色质修饰剂Prdm 3 和Prdm 16，表观遗传控制空间和时间的基因表达在颅面发育。的 该提案中概述的目标利用斑马鱼和小鼠的分子，遗传和表观遗传工具进行测试， Prdm 3和Prdm 16在Wn/β-catenin上游起作用以适当平衡软骨细胞假设 在颅面复合体的形成过程中的功能。在目标1中， 将定义斑马鱼颅面发育期间软骨细胞中的Wnt/β-连环蛋白转录活性。 通过Wnt/β-catenin的调节，软骨细胞极性和分化的保守功能将被阐明。 在哺乳动物颅面复合体中鉴定（目的2），最后，保守的Prdm 3-和 在颅面间充质中，Prdm 16调节的脊椎动物中的经典Wnt/β-连环蛋白增强子将 评估（目标3）。这些目标的完成将提供关于这些表观遗传修饰剂如何控制 特异性GRNs和信号传导模块（Wnt/β-catenin），以促进软骨形成， 颅面骨骼重要的是，该项目还将提供这些损失背后的机制见解， 这些因素有助于颅面疾病的发展。研究培训计划沿着职业生涯 本建议书中概述的发展和指导计划旨在为我的职业生涯奠定基础 目标是成为顶尖研究机构的独立研究员。在K99阶段，我将收到 克里斯汀·阿尔廷格（Kristin Artinger）在斑马鱼生物学方面的指导，以及成员们对我的建议的指导。 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 与促进赠款写作，科学交流，领导和指导技能的活动相一致， 与马赛克计划，将有助于我过渡到一个独立的教师职位在R 00阶段。 科罗拉多大学安舒茨医学院的颅面科提供了一个特殊的 环境与无数的资源来进行这项研究和培训计划，将指导我的成功 实现我的职业目标