The role of the Arp2/3 complex in cellular actin dynamics

Arp2/3 复合物在细胞肌动蛋白动力学中的作用

• 批准号: 8928640
• 负责人: JAMES E BEAR
• 金额: $ 37.05万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8928640
• 关键词: ATP Hydrolysis; Actins; Address; Biochemical; Biological; Biological Process; Cardiovascular Diseases; Cause of Death; Cell Adhesion; Cell physiology; Cells; Chemotaxis; Clinical; Complex; Cues; Cytoskeleton; Data; Disease; Epithelial; Fibrosis; Filament; G Actin; Genes; Genetic; Glia Maturation Factor; Health; Human; Image Analysis; Immune response; Intercellular Junctions; Intervention; Intrinsic factor; Knockout Mice; Knowledge; Life; Light; Mesenchymal; Methods; Microfluidics; Microscopy; Molecular; Morphogenesis; Neoplasm Metastasis; Normal Cell; Pathologic Processes; Pathway interactions; Phagocytosis; Physiological; Physiological Processes; Physiology; Play; Process; Property; Proteins; Publishing; Recycling; Regulation; Role; Staging; Structure; Tamoxifen; Testing; Therapeutic; Work; Wound Healing; base; cell motility; cell type; cellular imaging; cofilin; directional cell; human EMS1 protein; in vivo; intravital imaging; migration; novel; optogenetics; profilin; recombinase; reconstitution; transcriptome sequencing; tumor; vasodilator-stimulated phosphoprotein

DESCRIPTION (provided by applicant): The proper regulation of actin dynamics is essential for many biological processes such as wound healing, immune response and morphogenesis, and plays a significant role in pathological processes such as cancer metastasis and cardiovascular disease, the two leading causes of death in the developed world. Despite its widespread involvement in normal physiology and disease, efforts to target actin dynamics for therapeutic purposes are at an early stage and clearly require a deeper understanding of the processes involved. The Arp2/3 complex is a critical player in actin dynamics that generates branched actin arrays which are thought to be important for many cellular processes including cell migration, phagocytosis and cell adhesion. Using cells derived from a conditional Arp2/3 knockout mouse (Arpc2 gene), we propose to address several important questions for the field of actin dynamics: 1) How are Arp2/3-branched actin networks disassembled and dynamically turned over in cells? We have developed a new optogenetic method to control Arp2/3 function in cells with light that will allow us to dissect the de-branching pathway. 2) Is Arp2/3 required for actin-dependent processes such as directed migration, phagocytosis and cell-cell junction establishment? This will be addressed using a clean, genetic deletion approach in primary cells both ex vivo using live-cell imaging approaches and in vivo using multiphoton intravital imaging. 3) How do cells coordinate Arp2/3 and non-Arp2/3 actin pathways to produce optimal actin dynamics? Using our Arp2/3- deficient cells, we will interrogate the Arp2/3-independent pathways that partially compensate for its loss and study how Arp2/3-dependent and -independent pathway act in a coordinated pathway to produce optimal actin dynamics in cells.

描述（由申请人提供）：肌动蛋白动力学的适当调节对许多生物过程至关重要，如伤口愈合、免疫反应和形态发生，并在癌症转移和心血管疾病等病理过程中发挥重要作用，这是发达国家的两大死亡原因。尽管肌动蛋白广泛参与正常生理和疾病，但以治疗为目的的针对肌动蛋白动力学的努力尚处于早期阶段，显然需要对所涉及的过程有更深入的了解。Arp2/3复合体在肌动蛋白动力学中起关键作用，产生分支肌动蛋白阵列，这被认为对许多细胞过程很重要，包括细胞迁移、吞噬和细胞粘附。利用条件Arp2/3敲除小鼠（Arpc2基因）的细胞，我们提出解决肌动蛋白动力学领域的几个重要问题：1)在细胞中，Arp2/3分支肌动蛋白网络是如何分解和动态翻转的？我们已经开发了一种新的光遗传学方法来控制细胞中的Arp2/3功能，这将使我们能够解剖去分支途径。2)在诸如定向迁移、吞噬和细胞-细胞连接建立等动作蛋白依赖过程中是否需要Arp2/3 ？这将通过在原代细胞中使用活细胞成像方法和在体内使用多光子活体成像的清洁基因缺失方法来解决。3)细胞如何协调Arp2/3和非Arp2/3肌动蛋白通路以产生最佳的肌动蛋白动力学？使用我们的Arp2/3-缺陷细胞，我们将询问部分补偿其损失的Arp2/3独立途径，并研究Arp2/3依赖和独立途径如何在协调途径中发挥作用，以在细胞中产生最佳的肌动蛋白动力学。