The Role of the Spectraplakin Short-Stop in Cell Migration

Spectraplakin 短停在细胞迁移中的作用

• 批准号: 8382944
• 负责人: Derek Anthony Applewhite
• 金额: $ 8.73万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-10 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8382944
• 关键词: Actins; Address; Adhesions; Affect; Behavior; Biochemical; Biological; Biological Assay; Cancer Patient; Cell Adhesion; Cell Culture Techniques; Cell Line; Cell Polarity; Cell-Cell Adhesion; Cell-Matrix Junction; Cells; Cellular biology; Cessation of life; Complement; Cytoskeleton; Data; Databases; Developmental Biology; Disease; Disseminated Malignant Neoplasm; Drosophila genus; Event; Family; Family member; Gap Junctions; Goals; Homologous Gene; Invaded; Knowledge; Laboratories; Lead; Literature; Maintenance; Malignant Neoplasms; Microscopic; Microscopy; Microtubules; Modeling; Molecular; Neoplasm Metastasis; Organ; Physiological; Play; Population; Prevention; Process; Protein Family; Regulation; Research; Resolution; Role; System; Systems Analysis; Testing; Tissues; Traction; Work; base; cancer cell; cell motility; combinatorial; crosslink; design; fly; in vivo; in vivo Model; innovation; insight; member; metastatic process; migration; mortality; novel; novel therapeutics; prevent; therapeutic target; tissue culture; tissue/cell culture

DESCRIPTION (provided by applicant): Cell migration requires the dynamics of the actin-microtubule cytoskeleton and the regulation of both cell- matrix and cell-cell adhesion; however, there is a fundamental gap in our knowledge of how these networks are coordinated. This gap prevents a comprehensive understanding of the mechanisms that govern cell movements, including metastasis. The long term goal of this project is to determine how adhesion and the cytoskeleton are integrated during cell migration, focusing on how the Drosophila spectraplakin Short-stop and its mammalian counter-part MACF1/ACF7, model cytolinkers and integrators, coordinate these networks. Spectraplakins are excellent candidates for this coordination as they can physically cross-link actin and microtubules while playing a critical roles in the regulation o adhesion. Cell migration is a result of a synergistic relationship between these networks; therefore investigating molecules that potentially facilitate this cooperative interaction represens the next crucial step in understanding this process. Our central hypothesis is that molecules that coordinate actin-microtubule cross-linking and adhesion function as a nexus in the regulation of cell migration. This hypothesis has been formulated on the basis of data generated in the applicant's laboratory as well as mounting evidence present in the literature. The rationale for the proposed research is that metastasis represents a major cause of mortality in cancer patients. A more extensive understanding of cell migration will lead to the identification of futur therapeutic targets and ultimately a decrease in cancer related deaths. This hypothesis will be tested by two specific aims: 1) Determine how the spectraplakins Shot and MACF1 regulate cell migration; and 2) Determine the biological mechanisms that regulate their activity. To accomplish the first aim I will use a combination novel motile Drosophila cell lines and mammalian tissue culture cells with high resolution microscopy. To complement these studies analysis of Shot's role in cell migration will be carried out in an in vivo model of cell migration border cell migration. In the second aim biochemical and microscopic analysis will be used to determine whether Shot and MACF1 undergo an intramolecular conformational change. For this aim both cell biology based and developmental biology based assays will be used to determine the biological significance of this conformational change. The approach is innovative because it will utilize both cell culture models,Drosophila and mammalian, and an in vivo model of cell migration in a complementary manner. The proposed research is significant as its aim is to elucidate the combinatorial affects of adhesion and cytoskeletal dynamics during cell migration. This knowledge has the potential to advance the field of cell motility, and putatively uncover therapeutic targets for the prevention of metastasis.

描述（由申请人提供）：细胞迁移需要肌动蛋白微管细胞骨架的动力学以及细胞基质和细胞细胞粘附的调节；但是，我们对这些网络如何协调的了解存在一个根本的差距。该差距阻止了对控制细胞运动的机制，包括转移。该项目的长期目标是确定在细胞迁移过程中如何整合粘附和细胞骨架，重点介绍果蝇Spectraplakin短路及其哺乳动物反零件MACF1/ACF7，模型细胞链链球链球链球链将和整合器的模型。 Spectraplakins是这种协调的出色候选者，因为它们可以在调节O粘附中发挥关键作用，因此可以进行物理交联的肌动蛋白和微管。细胞迁移是这些网络之间协同关系的结果。因此，研究可能促进这种合作相互作用的分子代表了理解这一过程的下一个关键步骤。我们的中心假设是，在调节细胞迁移的调节中，协调肌动蛋白 - 微管交联和粘附功能的分子。该假设是根据申请人实验室中产生的数据以及文献中存在的越来越多的证据提出的。拟议研究的理由是转移代表了癌症患者死亡率的主要原因。对细胞迁移的更广泛的了解将导致鉴定未来的治疗靶标，并最终导致与癌症相关的死亡减少。该假设将通过两个具体的目的来检验：1）确定Spectraplakins射击和MACF1如何调节细胞迁移； 2）确定调节其活性的生物学机制。为了实现第一个目标，我将使用具有高分辨率显微镜的新型运动果蝇细胞系和哺乳动物组织培养细胞。为了补充这些研究的研究分析，将在细胞迁移的体内模型边界细胞迁移中进行细胞迁移中的作用。在第二个目标中，将使用生化和微观分析来确定SHOT和MACF1是否经历分子内构象变化。为此，基于细胞生物学的基于细胞生物学和基于发育生物学的测定将用于确定这种构象变化的生物学意义。该方法具有创新性，因为它将以互补方式使用细胞培养模型，果蝇和哺乳动物的模型以及细胞迁移的体内模型。拟议的研究非常重要，因为其目的是阐明细胞迁移过程中粘附和细胞骨架动力学的组合影响。这些知识有可能提高细胞运动领域，并揭示了预防转移的治疗靶标。