The Role of Osmotic Engine in Confined Migration

渗透引擎在密闭迁移中的作用

基本信息

批准号：
8875330
负责人：
Konstantinos Konstantopoulos
金额：
$ 39.46万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-06-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8875330
关键词：
Actins Actomyosin Adhesions Affect Anatomy Automobile Driving Biological Process Biomedical Engineering Cancer Biology Cell membrane Cell model Cell physiology Cells Cellular biology Complex Confined Spaces Cues Cytoskeletal Proteins Cytoskeleton Data Developmental Biology Dynein ATPase Extracellular Matrix F-Actin Health Image Immigration In Vitro Individual Integrins Ion Channel Ion Pumps Ion Transport Ions Kinesin Knowledge Length Liquid substance Measures Mechanics Mediating Membrane Microfluidic Microchips Microtubules Modeling Molecular Molecular Biology Molecular Models Molecular Motors Myosin ATPase Myosin Type II Pinocytosis Play Process Proteins Regulation Relative (related person)Role Signal Transduction Structure Surface Technology Testing Text Theoretical model Transport Vesicles Vesicle Water Width Work abstracting aquaporin 3 base cell motility in vivo interdisciplinary approach mathematical model migration molecular modeling neoplastic cell polymerization stem theories tool trafficking two-dimensional uptake voltage water channel

项目摘要

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Understanding the mechanisms of cell migration is a fundamental question in cell, developmental and cancer biology. Cell motility is influenced by a complex interplay among intracellular mechanics and signaling, and the physical cues of the microenvironment. Our knowledge on the mechanisms of cell migration stems primarily from in vitro studies using two- dimensional (2D) surfaces. Cell locomotion in 2D is driven by cycles of actin protrusion, integrin-mediated adhesion and myosin-dependent contraction. However, cells in vivo migrate within 3D extracellular matrices and through 3D pre-existing longitudinal channels created by various anatomic structures. Accumulating evidence suggests that the physical confinement affects the regulation and mechanisms of cell locomotion. For instance, myosin contractility and 1 integrin- dependent adhesion are dispensable in cell migration through confined spaces, which persists even when F-actin is disrupted. Thus, an alternative mechanism is at play for cells migrating in narrow microchannels. We recently proposed a new mechanism of cell motility in confined spaces that is described by the osmotic engine model (OEM). According to OEM, a cell migrating in a narrow channel establishes a spatial gradient of aquaporins (AQPs), ion channels and pumps in the cell membrane, so that there is a net inflow of water at the leading edge and a net outflow of water at the cell trailing edge. We hypothesize that cells can use different mechanisms (actomyosin-based and water permeation-based) depending on the physical cues of the microenvironment. We herein propose to develop an integrated experimental and theoretical approach to delineate the mechanisms of cell entry and migration in confined spaces. Experimental work will directly interact with theory and modeling throughout our proposed studies. In Aim 1, we propose to directly measure water uptake by cells migrating through confined spaces, and decipher the molecular mechanisms of water permeation and the role of mechanosensitive (MS) ion channels in confined migration. In conjunction with experimental work, we will develop a comprehensive molecular model of OEM integrating the roles of AQPs, membrane voltages and ion channels and pumps in cell migration in narrow channels. Because confined migration largely depends on microtubule (MT) dynamics, we will determine the role of MT molecular motors and their synergistic effects with actin polymerization in establishing AQP and ion channel polarization (Aim 2). In Aim 3, we will examine the process of cell entry into physically- constricted spaces using our microfluidic device, and explore the role of cytoskeletal proteins, adhesions and membrane components during this process. Taken together, we will decipher the physical and molecular basis of a fundamentally new mechanism governing cell entry and migration in confined spaces in which AQPs, ion pumps and MS channels play key roles using a multidisciplinary approach, involving state-of-the-art bioengineering, imaging, molecular biology tools along with mathematical modeling

在此处输入文本，这是您应用程序的新摘要信息。本节必须不超过30行文字。了解细胞迁移的机制是细胞，发育和癌症生物学中的一个基本问题。细胞运动性受细胞内力学和信号传导之间的复杂相互作用的影响，以及的物理提示微环境。我们对细胞迁移步骤的机制的了解主要是从体外研究中使用两种研究的知识尺寸（2D）表面。 2D中的细胞运动是由肌动蛋白，整联蛋白介导的粘合剂和肌球蛋白依赖性收缩。但是，体内细胞在3D细胞外材料内迁移，并通过3D预先存在由各种解剖结构产生的纵向通道。积累的证据表明身体限制会影响细胞运动的调节和机制。例如，肌球蛋白的收缩力和1整合素 - 依赖的粘附在细胞通过狭窄空间迁移中是可分配的，即使F-肌动蛋白为破坏了。这是针对狭窄的微通道迁移的细胞的另一种机制。我们最近提出了一个渗透发动机模型（OEM）描述的狭窄空间中细胞运动的新机制。根据 OEM，在狭窄通道中迁移的细胞建立了水通道（AQP），离子通道和泵的空间梯度在细胞膜中，以便在前缘有水的净流入，在电池尾部有水出口边缘。我们假设细胞可以使用不同的机制（基于肌动球蛋白和基于水渗透的机制）取决于微环境的物理提示。我们在这里提出的提议要开发综合的实验和描绘狭窄空间中细胞进入和迁移的机制的理论方法。实验工作将在我们提出的整个研究中，直接与理论和建模相互作用。在AIM 1中，我们建议直接测量细胞的水吸收通过狭窄的空间迁移，并破译了水渗透的分子机制机械敏感（MS）离子通道在受约束迁移中的作用。结合实验工作，我们将开发一个整合AQP，膜电压和离子通道的作用的OEM的综合分子模型和细胞迁移中的泵在狭窄的通道中。因为密闭迁移很大程度上取决于微管（MT）动力学，我们将确定MT分子电机及其在肌动蛋白聚合中的协同作用的作用建立AQP和离子通道极化（AIM 2）。在AIM 3中，我们将研究细胞进入物理的过程 - 使用我们的微流体设备狭窄的空间，并探索细胞骨架蛋白，粘合剂和膜的作用在此过程中的组件。综上在限制空间中迁移细胞进入和迁移的新机制，其中AQP，离子泵和MS频道播放使用多学科方法的关键角色，涉及最先进的生物工程，成像，分子生物学工具与数学建模一起