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

基本信息

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
项目类别：
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

项目摘要

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于许多生物学过程（例如伤口愈合，免疫反应和形态发生）至关重要，并且在病理过程中起着重要作用，例如癌症转移和心血管疾病，这是发达国家的两个主要死亡原因。尽管它广泛参与了正常的生理和疾病，但以治疗目的靶向肌动蛋白动力学的努力仍处于早期阶段，并且显然需要更深入地了解所涉及的过程。 ARP2/3复合物是肌动蛋白动力学中的关键参与者，该动力学产生了分支肌动蛋白阵列，这对于许多细胞过程很重要，包括细胞迁移，吞噬作用和细胞粘附。使用源自条件ARP2/3基因敲除小鼠（ARPC2基因）的细胞，我们建议解决肌动蛋白动力学领域的几个重要问题：1）ARP2/3分支的肌动蛋白网络如何拆卸并动态地转移到细胞中？我们已经开发了一种新的光遗传学方法来控制具有光的细胞中的ARP2/3功能，这将使我们能够剖析分支途径。 2）依赖肌动蛋白依赖性过程的ARP2/3（例如定向迁移，吞噬作用和细胞 - 细胞连接机构）是否需要？这将通过使用活细胞成像方法和使用Multiphoton经液室内成像在原代细胞中使用干净的遗传缺失方法来解决这一问题。 3）细胞如何协调ARP2/3和非ARP2/3肌动蛋白途径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ARP2/3独立的途径，该途径部分补偿了其损失，并研究ARP2/3依赖性和非依赖性途径如何在协调的途径中起作用，以在细胞中产生最佳的肌动蛋白动力学。