Role of the Ciliary Protein C2CD3 in Mandibular Skelotogenesis

睫状蛋白 C2CD3 在下颌骨发育中的作用

• 批准号: 10599872
• 负责人: Evan Cortez Brooks
• 金额: $ 4.17万
• 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: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10599872
• 关键词: Address; Affect; Animal Model; Bilateral; Biological Assay; Biology; Birds; Breeding; Cartilage; Cell Polarity; Cells; Cellular Membrane; Centrioles; Cilia; Classification; Cre driver; Data; Defect; Deposition; Development; Diagnosis; Disease; Distal; Dysmorphology; EXT1 gene; EXT2 gene; Elements; Embryo; Environment; Epithelium; Etiology; Exostoses; FGF8 gene; Face; Fibroblast Growth Factor; Gene Expression; Heparitin Sulfate; Histologic; Human; Image; Immunohistochemistry; In Situ Hybridization; Jaw; Mandible; Mandibular Prominence; Mesenchymal; Mesenchyme; Micrognathism; Microtubules; Modeling; Molecular; Mus; Mutation; Oral; Organelles; Osteochondrodysplasias; Osteogenesis; PEA3; Pain; Pathway interactions; Patients; Phenotype; Proteins; Quantitative Reverse Transcriptase PCR; Role; SU5402; Signal Pathway; Signal Transduction; Skeletal Development; Skeleton; Structure; Syndrome; Techniques; Testing; Therapeutic; Tissues; Training; Work; body system; bone; cartilaginous; chondrodysplasia; ciliopathy; cilium biogenesis; craniofacial; digital; experimental study; implantation; improved; mouse model; mutant; overexpression; repaired; second harmonic; skeletal; skeletogenesis; tumor

Project Summary/Abstract Primary cilia are non-motile, microtubule-based organelles that protrude from the cellular membrane to sense the cell’s external environment and to coordinate the transduction of multiple signaling pathways. Disruptions in the structure or function of primary cilia result in a class of disorders known as ciliopathies. The skeleton is often affected in ciliopathies as ciliopathy patients often present with osteochondrodysplasias (OCDPs) - anomalies of bone and cartilage - such as micrognathia and mandibular dysmorphology. Oral-facial-digital syndromes are a subset of ciliopathies that present with several OCDPs. There is considerable phenotypic overlap between subtypes of oral-facial-digital syndromes and among other skeletal ciliopathies, making them difficult to properly diagnose and differentiate from each other. The substantial degree of phenotypic overlap of ciliopathies that affect the skeleton combined with the lack of a cure for ciliopathic OCDPs make this an issue of significant biomedical concern. This proposed work focuses on understanding the requirement of the ciliary protein C2cd3 during the development of the mandibular skeleton by utilizing the avian C2CD3 mutant talpid2 (ta2) and the conditional C2cd3fl/fl mouse. C2cd3 localizes to the distal centrioles of primary cilia and is required for ciliogenesis. Mutations in C2cd3 result in the human ciliopathy Oral-facial-digital syndrome subtype 14. My preliminary data indicates that talpid2 embryos present with numerous mandibular OCDPs, such as bilateral Meckel’s chondrodysplasias and hypoplastic skeletal elements. There has yet to be an in-depth study as to how these phenotypes arise. In Aim 1, I will determine if the bilateral Meckel’s chondrodysplasias are exostoses - cartilage-capped bony tumors defined by lack of cellular polarity and ciliary extension, reduced Ext1/2 expression, and reduced heparan sulfate synthesis. ta2 embryos also possess increased FGF8 expression in the developing mandible, but it is not known if this leads to mandibular bone hypoplasia. For Aim 2, I will determine if expanded mandibular FGF8 signaling is sufficient to induce ta2 mandibular skeletal hypoplasia using bead implantations in the developing avian mandible. Lastly, C2cd3 is expressed ubiquitously throughout the developing embryo and it is not known if loss of C2cd3 in the craniofacial epithelium or mesenchyme leads to mandibular OCDP. For Aim 3, I will breed C2cd3fl/fl mice with the dHAND-Cre and Crect mice to conditionally delete C2cd3 in the mandibular mesenchyme and epithelium to determine if C2cd3 loss in either tissue type results in mandibular OCDP. This study is important for advancing our understanding of the cellular and molecular etiology of mandibular OCDPs in ciliopathic patients. A greater understanding of the developmental mechanisms of mandibular OCDPs in ciliopathies is necessary for further refined diagnosis and the development of less invasive therapeutic options.

项目概要/摘要 初级纤毛是非活动的、基于微管的细胞器，从细胞膜突出来感知 细胞的外部环境并协调多个信号通路的转导。中断 初级纤毛的结构或功能导致一类称为纤毛病的疾病。骷髅往往是 由于纤毛病患者经常出现骨软骨发育不良 (OCDP) - 异常，因此受到纤毛病的影响 骨骼和软骨 - 例如小颌畸形和下颌畸形。口-面-指综合征是一种 与多种 OCDP 一起出现的纤毛病的子集。之间存在相当大的表型重叠 口腔-面部-数字综合征的亚型以及其他骨骼纤毛病，使其难以正确治疗 相互诊断和鉴别。纤毛病的表型重叠程度很大 影响骨骼，加上纤毛病 OCDP 缺乏治愈方法，使得这个问题成为一个重要的问题 生物医学关注。这项拟议工作的重点是了解纤毛蛋白 C2cd3 的需求 在利用禽类 C2CD3 突变体 talpid2 (ta2) 和 有条件的 C2cd3fl/fl 小鼠。 C2cd3 定位于初级纤毛的远端中心粒，并且是 纤毛发生。 C2cd3 突变导致人类纤毛病口腔-面部-数字综合征亚型 14。 初步数据表明talpid2胚胎存在大量下颌OCDP，例如双侧下颌 梅克尔软骨发育不良和骨骼发育不良。目前还没有深入研究如何 这些表型出现。在目标 1 中，我将确定双侧梅克尔软骨发育不良是否为外生骨疣 - 软骨帽骨肿瘤的定义是缺乏细胞极性和纤毛延伸，Ext1/2 减少 表达，并减少硫酸乙酰肝素合成。 ta2 胚胎中 FGF8 表达也增加 发育中的下颌骨，但尚不清楚这是否会导致下颌骨发育不全。对于目标2，我会 使用以下方法确定扩大的下颌 FGF8 信号传导是否足以诱导 ta2 下颌骨骼发育不全 将珠子植入正在发育的鸟类下颌骨中。最后，C2cd3 在整个细胞中普遍表达。 发育中的胚胎，目前尚不清楚颅面上皮或间充质中 C2cd3 的缺失是否会导致 下颌 OCDP。对于目标 3，我将用 dHAND-Cre 和 Cect 小鼠培育 C2cd3fl/fl 小鼠，以有条件地 删除下颌间质和上皮中的 C2cd3，以确定任一组织类型中是否存在 C2cd3 丢失 结果导致下颌 OCDP。这项研究对于增进我们对细胞和细胞的理解非常重要 纤毛病患者下颌 OCDP 的分子病因学。对发展有更深入的了解 下颌 OCDP 在纤毛病中的作用机制对于进一步精细诊断和开发是必要的 的侵入性较小的治疗选择。