The Role of CARMIL2 in Cell Migration and Invasion

CARMIL2 在细胞迁移和侵袭中的作用

• 批准号: 8396161
• 负责人: MICHAEL Hunter LANIER
• 金额: $ 2.84万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8396161
• 关键词: Actins; Biochemical; Biological; Biological Assay; Cancer Patient; Cause of Death; Cells; Cellular Morphology; Data; Employee Strikes; Epithelial; Exhibits; Family member; Filament; Future; Goals; Hand; Human; Immigration; Label; Lead; Malignant Epithelial Cell; Malignant Neoplasms; Mesenchymal; Microfilaments; Migration Assay; Modeling; Molecular; Mutate; Nature; Neoplasm Metastasis; Patients; Phenotype; Play; Process; Protein Binding Domain; Protein Family; Proteins; RNA Cap-Binding Proteins; Recruitment Activity; Regulation; Role; Site; Structure; Testing; Time; Tissues; Tumor Cell Invasion; Vimentin; Wound Healing; Yeasts; base; cancer cell; cancer therapy; cell motility; fibrosarcoma; improved; migration; movie; mutant; neoplastic cell; polarized cell; polymerization; research study; scaffold; tumor; yeast two hybrid system

DESCRIPTION (provided by applicant): The main cause of death in patients with cancer is the invasion of body tissues by metastasizing tumor cells. Migration and invasion by tumor cells during metastasis require the coordination of cytoskeletal networks to form polarized structures, such as lamellipodia and invadopodia. Vimentin expression is a hallmark of the epithelial-mesenchymal transition in carcinoma cells and correlates strongly with invasive potential. Although recent studies demonstrate that vimentin regulates lamellipodia formation and invadopodia elongation, the mechanisms through which vimentin exerts influence over actin assembly are not well understood; however, CARMIL2 is a strong candidate to play a key role. CARMIL2 localizes to vimentin filaments in cells and regulates capping protein (CP), a key regulator of actin networks, in actin polymerization assays. CARMIL2 knockdown cells exhibit defective migration, diminished lamellipodial ruffling and macropinocytosis, and possess a striking multi-lamellipodia phenotype. How vimentin localization relates to CARMIL2 function and the importance of CP regulation by CARMIL2 for cell migration and invasion are not established. Additionally, CARMIL2 possesses a CARMIL homology domain (CHD) of unknown function. The high level of sequence similarity of the CHD among CARMIL family members suggests that it serves a conserved role. The central hypothesis of this proposal is that CARMIL2 regulates the polarity and dynamics of the actin network during cell migration and invasion by regulating CP and coordinating actin filament dynamics with the vimentin network. The following specific aims will test this hypothesis using molecular, cell biological, and biochemical approaches: 1) To test the importance of CP regulation by CARMIL2 for cell migration and invasion; 2) To test the importance of CARMIL2 localization to vimentin filaments for cell migration and invasion; 3) To identify proteins that interact with the conserved CHD of CARMIL2. The long-term objective of this project is to elucidate how cells assemble polarized cytoskeletal networks to drive cell migration and invasion during tumor metastasis. Completion of this proposal will provide an improved understanding of these mechanisms and will aid in the search for new cancer therapies. PUBLIC HEALTH RELEVANCE: The main cause of death in patients with cancer is the invasion of body tissues by metastasizing tumor cells. This proposal explores the molecular basis by which cells polarize and assemble cytoskeletal networks to drive migration and invasion during tumor metastasis. Completion of this proposal will provide an improved understanding of metastasis and aid in the search for new cancer therapies.

描述（由申请方提供）：癌症患者死亡的主要原因是转移性肿瘤细胞侵入身体组织。肿瘤细胞在转移过程中的迁移和侵袭需要细胞骨架网络的协调以形成极化结构，如板状伪足和侵袭伪足。波形蛋白表达是癌细胞上皮-间质转化的标志，并与侵袭潜力密切相关。虽然最近的研究表明，波形蛋白调节板状伪足的形成和侵入伪足的伸长，波形蛋白通过其对肌动蛋白组装施加影响的机制还没有很好地理解;然而，CARMIL 2是一个强有力的候选人发挥关键作用。在肌动蛋白聚合测定中，CARMIL 2定位于细胞中的波形蛋白丝并调节加帽蛋白（CP），其是肌动蛋白网络的关键调节因子。CARMIL 2敲除细胞表现出迁移缺陷、板状伪足皱褶减少和巨胞饮作用，并具有显著的多板状伪足表型。波形蛋白定位如何与CARMIL 2功能相关以及CARMIL 2对CP调节对细胞迁移和侵袭的重要性尚未确定。此外，CARMIL 2还具有一个功能未知的CARMIL同源结构域（CHD）。在CARMIL家族成员中CHD的高水平序列相似性表明其具有保守的作用。该提议的中心假设是CARMIL 2通过调节CP和协调肌动蛋白丝动态与波形蛋白网络来调节细胞迁移和侵袭期间肌动蛋白网络的极性和动态。以下具体目标将使用分子、细胞生物学和生物化学方法来检验该假设：1）检验CARMIL 2对CP调节对于细胞迁移和侵袭的重要性; 2）检验CARMIL 2定位于波形蛋白丝对于细胞迁移和侵袭的重要性; 3）鉴定与CARMIL 2的保守CHD相互作用的蛋白质。本项目的长期目标是阐明细胞如何组装极化的细胞骨架网络，以驱动肿瘤转移过程中的细胞迁移和侵袭。该提案的完成将使人们更好地了解这些机制，并有助于寻找新的癌症疗法。 公共卫生相关性：癌症患者死亡的主要原因是转移性肿瘤细胞侵入身体组织。这一提议探讨了细胞在肿瘤转移过程中形成和组装细胞骨架网络以驱动迁移和侵袭的分子基础。该提案的完成将提供对转移的更好理解，并有助于寻找新的癌症治疗方法。