The Role of SPECC1L cytoskeletal protein in craniofacial development and malformation

SPECC1L细胞骨架蛋白在颅面发育和畸形中的作用

基本信息

批准号：
10213181
负责人：
Irfan Saadi
金额：
$ 22.71万
依托单位：
UNIVERSITY OF KANSAS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10213181
关键词：
Adherens Junction Adhesions Affect Alleles Attenuated Bilateral Biological Assay Branchial arch structure Cell Adhesion Cell Communication Cell Density Cell Polarity Cell membrane Cell physiology Cells Cephalic Chemicals Collaborations Complex Congenital Abnormality Craniofacial Abnormalities Cultured Cells Cytoskeletal Proteins Data Defect Destinations Disease Embryo Embryonic Development Epithelial Epithelium Exencephalies Exhibits Eye Face Family Future Genes Genetic Goals Human Image In Vitro Intercellular Junctions Iowa Japan Knowledge Link Live Birth Measures Mediating Mediation Medical Care Costs Membrane Mesenchymal Modeling Molecular Morphogenesis Movement Mus Mutate Mutation Neural Crest Neural Crest Cell Neural Fold Neural Tube Closure Neural tube Neuroectoderm Neuroepithelial Neuroepithelial Cells Opitz syndrome Oral cavity Orbital separation excessive Palate Pathogenesis Pathway interactions Patients Perinatal Persons Phenotype Play Prevention strategy Productivity Proto-Oncogene Proteins c-akt Role Sampling Services Severities Signal Transduction Stream Syndrome Teebi syndrome Testing Therapeutic Tissues Transmission Electron Microscopy Ubiquitination base cell behavior cell motility cleft lip and palate craniofacial craniofacial development de novo mutation density dosage improved in vivo inhibitor/antagonist insight life time cost malformation migration mouse model mutant neural plate new therapeutic target novel oral cavity epithelium orofacial cleft palatal shelves palatogenesis perinatal period postnatal small molecule surgery outcome

项目摘要

PROJECT SUMMARY Orofacial clefts are among the most common birth defects in the U.S., occurring in 1/800 live-births. The lifetime cost for medical treatment, educational services and lost productivity averages more than $100,000 per affected person. While a number of contributory genes have been identified, there is a continued need to understand the underlying pathogenetic mechanisms. We identified the first de novo mutations in a novel cytoskeletal SPECC1L gene in patients with oblique facial clefts. Recent identification of SPECC1L mutations in patients with syndromic and nonsyndromic cleft lip and palate suggests that insights into the cellular and molecular mechanism of SPECC1L function will be directly relevant to both severe and common facial malformations. Our data show that Specc1l is expressed in the neural folds at E8.5, including in pre-migratory cranial neural crest cells (CNCCs). Later at E9.5, it is expressed in post-migratory CNCCs, and in the branchial arches at E10.5. To study SPECC1L function, we have generated severe, hypomorphic and conditional mouse alleles of Specc1l deficiency. The severe mouse allele is embryonic lethal at E9.5 with defective neural tube (NT) closure and incomplete delamination of post-migratory CNCCs. The hypomorphic allele shows incompletely penetrant perinatal exencephaly phenotype. Moderate mutants with one hypomorphic and one severe allele show highly penetrant palate closure delay. In addition, SPECC1L-deficient cells show altered adherens junctions (AJs) and reduced PI3K-AKT signaling, both in vitro in cultured cells and in vivo in Specc1l mutant tissue. Modulation of cell-cell contacts is important not only for CNCC delamination from the neuroectoderm, but also for “collective” migration of CNCCs to their defined destinations. The central hypothesis of this project is that SPECC1L modulates cell adhesion and collective migration by regulating the density of epithelial and mesenchymal cell-cell contacts through PI3K-AKT signaling. In Aim 1, we propose to investigate the effect of Specc1l dosage on craniofacial development using our hypomorphic, severe, and conditional mouse alleles. In Aim 2, we will investigate SPECC1L mediation of AJ stability and PI3K-AKT signaling using small molecule modulators of PI3K-AKT pathway. We will also explore the mechanism underlying SPECC1L mediation of AKT stability. In Aim 3, we will use live-imaging to quantitatively assess changes in collective cell behavior. Analyses will be conducted in ex vivo cultured E8.5 neural plate explants, with CNCCs marked with Wnt1-Cre or Sox10-Cre. Successful completion of these studies will establish novel SPECC1L-based links between AJs, cell polarity and PI3K-AKT signaling during facial morphogenesis and palate closure. Understanding the correlation between SPECC1L dosage or functional deficiency and collective cell migration will provide targets for future therapeutic or preventative strategies against orofacial clefting.

项目概要口面部裂是美国最常见的出生缺陷之一，发生在 1/800 的活产儿中。医疗、教育服务和生产力损失的终生成本平均每人超过 100,000 美元虽然已经确定了许多贡献基因，但仍需要继续研究。我们了解了潜在的致病机制。斜面裂患者的细胞骨架 SPECC1L 基因最近鉴定的 SPECC1L 突变。在患有综合征性和非综合征性唇腭裂的患者中，表明对细胞和 SPECC1L功能的分子机制将与严重和普通面部直接相关我们的数据显示 Specc1l 在 E8.5 的神经褶皱中表达，包括在迁移前的神经褶皱中表达。后来在 E9.5，它在迁移后的 CNCC 和脑神经嵴细胞中表达。为了研究 SPECC1L 功能，我们生成了严重的、低等位的鳃弓。 Specc1l 缺陷的条件性小鼠等位基因，严重的小鼠等位基因在 E9.5 时是胚胎致死的。神经管 (NT) 闭合缺陷和迁移后 CNCC 的不完全分层。等位基因显示不完全渗透的围产期无脑畸形表型，具有一种。此外，SPECC1L 缺陷的低等位基因和一个严重的等位基因表现出高度渗透性的腭闭合延迟。在体外培养的细胞和在体内 Specc1l 突变组织中，细胞-细胞接触的调节不仅对于 CNCC 分层很重要。从神经外胚层，也可以将 CNCC“集体”迁移到其指定的目的地。该项目的假设是 SPECC1L 通过调节细胞粘附和集体迁移在目标 1 中，我们建议通过 PI3K-AKT 信号传导上皮细胞和间质细胞接触的密度。使用我们的低等位、严重和在目标 2 中，我们将研究 SPECC1L 对 AJ 稳定性和 PI3K-AKT 的介导。我们还将探索使用 PI3K-AKT 途径的小分子调节剂进行信号传导的机制。在目标 3 中，我们将使用实时成像来定量评估 AKT 稳定性。集体细胞行为的变化将在离体培养的 E8.5 神经板外植体中进行。成功完成这些研究将建立带有 Wnt1-Cre 或 Sox10-Cre 标记的 CNCC。面部形态发生过程中 AJ、细胞极性和 PI3K-AKT 信号之间基于 SPECC1L 的联系了解 SPECC1L 剂量或功能缺陷与上颚闭合之间的相关性。集体细胞迁移将为未来口面部的治疗或预防策略提供目标裂开。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Novel insights into the fundamentals of palatal shelf elevation dynamics in normal mouse embryos.

对正常小鼠胚胎腭架抬高动力学基本原理的新见解。

DOI：
发表时间：
2022-05
期刊：
影响因子：
0
作者：
Saadi, Irfan;Goering, Jeremy;Moedritzer, Michael;Stetsiv, Marta;Isai, Dona Greta;Hufft;Umar, Zaid;Rickabaugh, Madison K;Keselman, Paul;Chauhan, Munish;Brooks, William;Fischer, Kenneth;Czirok, Andras
通讯作者：
Czirok, Andras

Phenotypic spectrum associated with SPECC1L pathogenic variants: new families and critical review of the nosology of Teebi, Opitz GBBB, and Baraitser-Winter syndromes.

与 SPECC1L 致病变异相关的表型谱：Teebi、Opitz GBBB 和 Baraitser-Winter 综合征的新家族和分类学严格审查。

DOI：
发表时间：
2019-12
期刊：
影响因子：
0
作者：
Bhoj, Elizabeth J;Haye, Damien;Toutain, Annick;Bonneau, Dominique;Nielsen, Irene Kibæk;Lund, Ida Bay;Bogaard, Pauline;Leenskjold, Stine;Karaer, Kadri;Wild, Katherine T;Grand, Katheryn L;Astiazaran, Mirena C;Gonzalez;Carvalho, An
通讯作者：
Carvalho, An