Mechanism of Cranial Neural Crest Cell Migration

颅神经嵴细胞迁移机制

基本信息

批准号：
8441477
负责人：
DOMINIQUE R ALFANDARI
金额：
$ 37万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-03-01 至 2017-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8441477
关键词：
Adhesives Amino Acid Sequence Animal Model Autoimmune Diseases Behavior Binding Bioinformatics C-terminal Cell Adhesion Cell Adhesion Molecules Cell Cycle Progression Cell Nucleus Cell Proliferation Cell membrane Cell surface Cells Cephalic Cleaved cell Cytoplasmic Tail Cytoskeleton DNA Modification Process Data Defect Development Developmental Cell Biology Disintegrins E-Cadherin EGF gene Embryo Embryology Embryonic Development Ephrins Epithelium Extracellular Domain Face Family Fibroblast Growth Factor Fluorescence Resonance Energy Transfer Funding Gene Expression Genes Genetic Transcription Goals Guanine Guanine Nucleotide Exchange Factors Heart Immigration In Vitro Knowledge Light Link Liver Malignant Neoplasms Mediating Mesenchyme Metalloproteases Microscopy Modeling Molecular Models Monomeric GTP-Binding Proteins N-terminal Neoplasm Metastasis Nerve Degeneration Neural Crest Neural Crest Cell Pathologic Processes Pathology Pathway interactions Physiological Processes Population Process Proteins Public Health Regulation Research Role Signal Pathway Signal Transduction Signaling Molecule Site Skeletal Muscle Stem cells Structure Surface System TNF gene Techniques Testing Time Tissues base bone cadherin-11 cancer cell cell motility cohesion craniofacial design extracellular human disease in vitro Assay in vivo innovation intercellular communication member migration molecular modeling notch protein overexpression promoter receptor research study rho transcription factor

项目摘要

DESCRIPTION (provided by applicant): The migration of cranial neural crest (CNC) cells is controlled by proteins that regulate gene expression, cell proliferation, cell signaling and cell adhesion. Among these proteins the ADAM cell surface metalloproteases can regulate both cell signaling and cell adhesion. They do so by cleaving signaling and adhesion molecules from the cell surface. There is a fundamental gap in understanding how ADAM proteins control cell migration in general and CNC migration in particular. Our long-term goal is to understand how CNC migration is controlled in a developing embryo. The objective of this application is to determine how ADAM metalloproteases control CNC migration. The central hypothesis of this proposal, based on extensive preliminary data, is that ADAM metalloproteases control cell migration both by shedding cell adhesion molecules from the plasma membrane and by the independent activity of their cytoplasmic domains inside of the nucleus. Aim 1: To identify how ADAM cleavage of Cadherin-11 controls CNC migration. We have shown that Cad-11 is cleaved by ADAM13 during CNC migration and that the Cad-11 extracellular fragment containing the adhesive site is shed and can rescue migration in embryos lacking ADAM13 or overexpressing Cad-11. Our hypothesis is that this fragment binds to the surface of CNC via a receptor and promotes migration. We propose to determine how the extracellular fragment of Cad-11 promotes migration. We will determine, 1) what are the consequences of not cleaving Cad-11 on CNC specification and migration, 2) if the Cad-11 extracellular fragment stimulates directed cell migration and/or interfere with contact mediated inhibition, and 3) What are the signaling pathways involved in this process. Aim 2: To identify the mechanism by which the ADAM cytoplasmic domain controls CNC migration. We have shown that the ADAM13 cytoplasmic domain (C13) is cleaved and translocates into the nucleus. We showed that this is critical for gene expression and CNC migration in vivo. We also showed that one gene regulated by C13, Calpain8 is essential for CNC migration. Our hypothesis is that the ADAM13 cytoplasmic domain controls gene expression by modifying the function of a transcription factor to promote CNC migration. We will determine 1) the amino acid sequence(s) in C13 that are responsible for the control of gene expression and cell migration, and 2) the mechanism by which C13 regulates the expression of Calpain8. The approach is innovative, because it shows for the first time in any system that ADAM cytoplasmic domains function in the nucleus to regulate specific gene expression and cell migration. The proposed research is significant, because it is expected to vertically advance the field by identifying evolutionary conserved motifs in ADAM cytoplasmic domain that control the activity of transcription factors (e.g. FoxD3). In addition, the Cad-11 extracellular domain has been shown to increase cancer cell invasion, thus our results will shed the light on the mechanism by which this domain operates.

描述（由申请人提供）：颅神经rest（CNC）细胞的迁移受调节基因表达，细胞增殖，细胞信号传导和细胞粘附的蛋白质的控制。在这些蛋白质中，亚当细胞表面金属蛋白酶可以调节细胞信号传导和细胞粘附。他们通过从细胞表面切割信号传导和粘附分子来做到这一点。了解亚当蛋白如何控制细胞迁移，尤其是CNC迁移存在基本差距。我们的长期目标是了解如何在发育中的胚胎中控制CNC迁移。该应用的目的是确定亚当金属蛋白酶如何控制CNC迁移。基于广泛的初步数据，该提案的中心假设是，亚当金属蛋白酶通过从质膜中脱离细胞粘附分子和核内部细胞质结构域的独立活性来控制细胞迁移。目的1：确定亚当111的裂解如何控制CNC迁移。我们已经表明，CAD-11在CNC迁移过程中被ADAM13裂解，并且含有粘合剂位点的CAD-11细胞外片段被脱落，并且可以挽救缺乏ADAM13或过度表达CAD-111的胚胎中的迁移。我们的假设是，该片段通过受体与CNC的表面结合并促进迁移。我们建议确定CAD-11的细胞外碎片如何促进迁移。我们将确定，1）不裂解CAD-11对CNC规范和迁移的后果是什么，2）如果CAD-11细胞外片段刺激了定向的细胞迁移和/或干扰接触介导的抑制，而3）3）涉及的信号通路是什么。目标2：确定亚当细胞质结构域控制CNC迁移的机制。我们已经表明，ADAM13细胞质结构域（C13）被裂解并易位到细胞核中。我们表明，这对于体内基因表达和CNC迁移至关重要。我们还表明，一个受C13调节的基因，CalPain8对于CNC迁移至关重要。我们的假设是ADAM13细胞质结构域通过修改转录因子的功能以促进CNC迁移来控制基因表达。我们将确定1）C13中负责控制基因表达和细胞迁移的氨基酸序列，以及2）C13调节CALPAIN8的表达的机制。该方法具有创新性，因为它在任何系统中首次显示亚当细胞质结构域在细胞核中起作用以调节特定基因表达和细胞迁移。拟议的研究很重要，因为预计通过鉴定控制转录因子活性的亚当细胞质结构域中的进化保守基序，可以垂直垂直提高该领域（例如FOXD3）。此外，CAD-11的细胞外结构域已被证明会增加癌细胞的侵袭，因此我们的结果将阐明该结构域运行的机制。