Mechanism of Cranial Neural Crest Cell Migration

颅神经嵴细胞迁移机制

• 批准号: 8236674
• 负责人: DOMINIQUE R ALFANDARI
• 金额: $ 37.09万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 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/8236674
• 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

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. PUBLIC HEALTH RELEVANCE: The proposed project is relevant to public health because it increases our knowledge of craniofacial development in a vertebrate model organism amenable to experimental studies. The understanding of ADAM control of cell migration is also relevant as these proteins are often up-regulated and associated with invasion of cancer cells. The current study also shows that ADAM cytoplasmic domains control the expression of genes that are relevant to multiple human diseases.

描述（由申请方提供）：颅神经嵴（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控制。在这些蛋白质中，ADAM细胞表面金属蛋白酶可以调节细胞信号传导和细胞粘附。它们通过从细胞表面切割信号和粘附分子来实现这一点。在理解ADAM蛋白如何控制细胞迁移，特别是CNC迁移方面存在根本性的差距。我们的长期目标是了解CNC迁移如何在发育中的胚胎中控制。本申请的目的是确定ADAM金属蛋白酶如何控制CNC迁移。基于广泛的初步数据，该提议的中心假设是ADAM金属蛋白酶通过从质膜脱落细胞粘附分子和通过其细胞质结构域在核内的独立活性来控制细胞迁移。目的1：确定钙粘蛋白-11的ADAM切割如何控制CNC迁移。我们已经表明，Cad-11被切割的ADAM 13 CNC迁移过程中，Cad-11细胞外片段含有的粘合剂网站脱落，可以挽救缺乏ADAM 13或过度表达Cad-11的胚胎迁移。我们的假设是，该片段通过受体与CNC表面结合并促进迁移。我们建议确定Cad-11的细胞外片段如何促进迁移。我们将确定，1）不切割Cad-11对CNC特化和迁移的影响，2）Cad-11胞外片段是否刺激定向细胞迁移和/或干扰接触介导的抑制，以及3）参与该过程的信号传导途径是什么。目的2：确定ADAM胞质结构域控制CNC迁移的机制。我们已经证明，ADAM 13胞质结构域（C13）被切割并易位到细胞核中。我们发现，这对体内基因表达和CNC迁移至关重要。我们还发现，一个由C13调控的基因，Calpain 8是CNC迁移所必需的。我们的假设是，ADAM 13的胞质结构域控制基因的表达，通过修改的转录因子的功能，以促进CNC迁移。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Calpain 8表达的机制。该方法是创新的，因为它首次在任何系统中表明，ADAM胞质结构域在细胞核中发挥作用，以调节特定的基因表达和细胞迁移。这项研究意义重大，因为它有望通过鉴定ADAM胞质结构域中控制转录因子（如FoxD 3）活性的进化保守基序来垂直推进该领域。此外，Cad-11胞外结构域已被证明可以增加癌细胞的侵袭，因此我们的研究结果将揭示该结构域的运作机制。 公共卫生相关性：拟议的项目是有关公共卫生，因为它增加了我们的知识颅面发育的脊椎动物模式生物适合实验研究。对细胞迁移的ADAM控制的理解也是相关的，因为这些蛋白质通常被上调并与癌细胞的侵袭相关。目前的研究还表明，ADAM胞质结构域控制着与多种人类疾病相关的基因的表达。