Role of Cytoskeletal Protein SPECC1L in Facial Morphogenesis and Facial Clefting

细胞骨架蛋白 SPECC1L 在面部形态发生和面部裂隙中的作用

• 批准号: 8480396
• 负责人: Irfan Saadi
• 金额: $ 22.65万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8480396
• 关键词: Actins; Affect; Alleles; Antibodies; Biological Assay; Branchial arch structure; Cell Line; Cell-Cell Adhesion; Cells; Cellular Morphology; Cephalic; Cleaved cell; Cleft lip with or without cleft palate; Collaborations; Complement; Congenital Abnormality; Cytoskeletal Proteins; Cytoskeleton; Data; Defect; Development; Drosophila genus; ES Cell Line; Employee Strikes; Endothelin; Exhibits; Eye; Face; Focal Adhesions; Galactosidase; Genes; Genomics; German population; Homologous Gene; Human; Image; Instruction; Integrin Signaling Pathway; Integrins; Kinetics; Label; Life; Live Birth; Medical; Microtubules; Modeling; Molecular; Morphogenesis; Mus; Mutate; Mutation; Neural Crest; Neural Crest Cell; Nuclear; Oral cavity; Orthologous Gene; Pathway interactions; Patients; Persons; Phenocopy; Primordium; Productivity; Proteomics; Publications; Regulation; Reporter; Reporter Genes; Role; Screening procedure; Services; Signal Pathway; Signal Transduction; Speed; Staging; Structure; Testing; Tissues; Transgenic Organisms; Tubulin; Tungsten; Vertebrates; Zebrafish; base; cDNA Arrays; cell motility; cellular imaging; crosslink; depolymerization; fly; insight; knock-down; life time cost; link protein; migration; mouse model; mutant; novel; orofacial; polymerization; protein protein interaction; vector

PROJECT SUMMARY (See instructions): Orofacial clefts are one of the most common birth defects in the U.S., occurring in 1/750 live births. The lifetime cost for medical treatment, educational services and lost productivity averages more than $100,000 per affected person. While the majority of orofacial clefts result in cleft lip with or without cleft palate (CL/P), a small percentage results in oblique facial clefts (ObFC) that extend from the oral cavity to the eye. Although less common, insights into the cellular mechanism of ObFC - first definitively classified by Paul Tessier in 1976 - have remained elusive. We have identified two de novo occurrences of SPECC1L mutations in patients with ObFC. Our studies in zebrafish and fly provide significant insight into SPECC1L function, which thus far had remained unstudied with no scientific publications. Knockdown of a previously uncharacterized zebrafish SPECC1L homolog perturbs cranial neural crest (CNC) and results in a dramafic loss of facial structures, thus extending SPECC1L function in facial morphogenesis to other vertebrates. In addition, knockdown of the sole uncharacterized Drosophila ortholog phenocopies - to an extraordinary extent - known fly mutants in the integrin-signaling pathway that exhibit cell adhesion and migration defects. Furthermore, our cellular and molecular analyses show that SPECC1L is a novel cytoskeletal cross-linking protein that interacts with both the microtubule and actin cytoskeletons. Transient expression of SPECC1LGFP stabilizes a subset of microtubules, while SPECC1L knockdown causes defective actin cytoskeleton reorganization and impairs cell adhesion and migration. Together with mouse Speed I expression in the developing facial prominences, these results begin to explain how human ObFC can arise following SPECC1L deficiency. The aim of this proposal is to develop a mouse model to test the pathogenetic mechanism of SPECC1L deficiency in mammalian facial morphogenesis (Aim 1) and to precisely define the cellular (Aim 2) and molecular (Aim 3) role of SPECC1L in CNC cell migration and specification of facial structures.

项目摘要（请参阅说明）： 口面裂缝是美国最常见的先天缺陷之一，发生在1/750 Live出生中。医疗治疗，教育服务和生产力损失的终生成本平均每个受影响的人超过100,000美元。虽然大多数口面裂口会导致唇lip裂，有或不带left裂（Cl/p），但较小的百分比导致倾斜的面部裂口（OBFC）从口腔腔延伸到眼睛。 尽管不常见，但对OBFC的细胞机制的见解 - 首先是由Paul Tessier于1976年定义的 - 仍然难以捉摸。我们已经确定了OBFC患者中SPECC1L突变的两个从头出现。我们在斑马鱼和Fly方面的研究提供了对SpecC1L功能的重要见解，到目前为止，该功能尚未研究，没有科学出版物。敲除先前未表征的斑马鱼Specc1l同源物颅神经Crest（CNC），并导致戏剧性 面部结构的丧失，从而将面部形态发生中的Specc1l功能扩展到其他脊椎动物。此外，在整合素信号途径中，唯一未表征的果蝇直系同源物表现出了唯一的未表征的果蝇直系同源表膜，这些蝇素突变体表现出细胞粘附和迁移缺陷。 此外，我们的细胞和分子分析表明，SPECC1L是一种新型的细胞骨架交联蛋白，与微管和肌动蛋白细胞骨架相互作用。 SpecC1LGFP的瞬时表达稳定了微管的子集，而SPECC1L敲低会导致肌动蛋白细胞骨架的不良重组，并损害细胞粘附和迁移。与小鼠速度I在发育中的面部突出中的表达一起，这些结果开始解释人类OBFC在SPECC1L缺乏之后如何出现。该建议的目的是开发小鼠模型，以测试SPECC1L缺乏症在哺乳动物面部形态发生中的致病机制（AIM 1），并精确定义细胞（AIM 2）和SPECC1L在CNC细胞迁移和面部结构规范中SPECC1L的作用。