Tracing the origins of craniofacial growth plates

追踪颅面生长板的起源

• 批准号: 10413816
• 负责人: Lindsey Anne Barske
• 金额: $ 15.9万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10413816
• 关键词: ATAC-seq; Adult; Affect; Birth; Cartilage; Cell Line; Cell Polarity; Cells; Characteristics; Chick; Chondrocytes; Craniofacial Abnormalities; Cre lox recombination system; Data; Development; Dorsal; Embryo; Enhancers; Epiphysial cartilage; Face; Future; Gene Expression; Genes; Growth; Head; Human; Image; Joint structure of suture of skull; Joints; Label; Location; Logic; Maintenance; Mammals; Maps; Methodology; Methods; Modeling; Molecular; Mus; National Institute of Dental and Craniofacial Research; Neural Crest; Pathway interactions; Pattern; Physiologic Ossification; Pilot Projects; Population; Positioning Attribute; Prevalence; Process; Property; RNA; Reagent; Reporter; Research; Research Design; Shapes; Site; Skeletal system; Skeleton; Specific qualifier value; Specificity; Surgical sutures; Testing; Tissues; Transgenes; Transgenic Organisms; Work; Zebrafish; apoAI regulatory protein-1; bone; cartilaginous; clinically relevant; cohort; craniofacial; craniofacial bone; craniofacial tissue; cranium; emx2 protein; experimental study; innovation; intein; limb bone; long bone; novel; osteoprogenitor cell; postnatal; prevent; promoter; reconstitution; single-cell RNA sequencing; skeletal; skeletal dysplasia; stem cells; transcription factor

PROJECT SUMMARY Growth of the skeleton occurs at specialized sites called growth plates and sutures. Where these growth sites are located and how long they persist postnatally determine the final shape and size of the skeleton. The prevalence of skeletal dysplasias affecting bone or cartilage growth in humans (estimated to impact approximately one in 4,000-5,000 births) has motivated a vast body of research into the pathways regulating growth plate and suture development and function. However, the field still lacks a basic understanding of the mechanisms that determine where these critical sites form within the skeleton. Growth plates in particular have been typically assumed to form passively between centers of endochondral ossification. However, a scattering of cell polarity data from zebrafish, mice, and chick indicate that growth plates may be prefigured in the cartilage template well before ossification begins, suggesting an active patterning process. This application presents a novel hypothesis for growth plate origins, to be tested in zebrafish using an innovative fate-mapping strategy newly applied to the skeletal system. Growth plates are present in endochondral bones of the craniofacial skeleton as well as the long bones of the limbs. Prior to the onset of bone or cartilage differentiation in the head, skeletal progenitors for the face express distinct cohorts of genes depending on their position on the dorsal-ventral (DV) axis. These domains are not entirely mutually exclusive: a line of cells co-expressing markers characteristic of the neighboring domains can typically be detected at each boundary. Importantly, the positions of these boundaries appear to correlate with the eventual distribution of growth plates and cartilaginous joints in the adult skull. This observation prompted the hypothesis that cartilaginous growth zones in the skull are destined to form at these molecular boundaries. Testing this hypothesis requires following the fate of embryonic boundary cells to adulthood. To this end, an underutilized but powerful intersectional split-Intein-Cre lineage-tracing method will be used to specifically label cells co-expressing markers of adjacent DV domains. The pilot study outlined here will provide the first test of this original hypothesis, validate the methodology, and create reagents for future extensions of this work, all prerequisites for future R01-level support. This proposal directly aligns with the NIDCR's call to investigate mechanisms of development, maintenance, and remodeling of craniofacial tissues. Though the scope is currently limited to the skull, the principles under study may also apply to many other parts of the endochondral skeleton and possibly also to cranial sutures. Results from this work could stimulate a major advance in understanding the logic governing skeletal patterning, with clear relevance for human craniofacial malformations and other skeletal dysplasias.

项目摘要 骨骼的生长发生在称为生长板和缝合线的专用地点。这些成长的地方 位置位于位置，以及它们产后确定骨骼的最终形状和大小多长时间。 影响人类骨骼或软骨生长的骨骼发育不良的患病率（估计会影响 大约有4,000-5,000分的分娩）激发了对规范的路径的大量研究 生长板和缝合板的发展和功能。但是，该领域仍然缺乏对 确定这些关键位点在骨骼内形成的机制。特别是生长板 通常假定在内侧软骨骨化中心之间被动形成。但是， 斑马鱼，小鼠和雏鸡的细胞极性数据的散射表明生长板可能被预先构成 在骨化开始之前，在软骨模板中，提出了一个主动的模式过程。这 应用提出了一种新的假设生长板起源的假设，可以在斑马鱼中使用创新进行测试 命运策略新应用于骨骼系统。 生长板存在于颅面骨骼的内骨骨骼以及长骨 四肢。在头部骨头或软骨分化开始之前，面部的骨骼祖细胞 根据其在背腹侧（DV）轴上的位置表达不同的基因队列。这些域 不是完全相互排斥的：相邻的一系列共同表达标记的特征 通常可以在每个边界检测到域。重要的是，这些边界的位置出现 与成人头骨中生长板和软骨关节的最终分布相关。这 观察促使一个假设，即颅骨中的软骨生长区注定要形成 这些分子边界。检验该假设需要遵循胚胎边界细胞的命运 成年。为此，一种未充分利用但功能强大的交叉分裂谱系谱系追踪方法 将用于特异性标记相邻DV域共表达标记的细胞。试点研究 此处概述将提供对该原始假设的首次测试，验证方法，并创建 这项工作的未来扩展的试剂，这是未来R01级支持的所有先决条件。 该提案直接与NIDCR调查开发，维护机制， 和颅面组织的重塑。尽管范围目前仅限于头骨，但 研究还可能适用于内软骨骨骼的许多其他部位，也可能适用于颅缝线。 这项工作的结果可能会刺激理解控制骨骼的逻辑的重大进步 具有图案，与人类颅面畸形和其他骨骼发育不良相关。