Viscotaxis: Novel cell migration mechanisms regulated by microenvironmental viscosity

Viscotaxis：微环境粘度调节的新型细胞迁移机制

基本信息

批准号：
10379292
负责人：
Konstantinos Konstantopoulos
金额：
$ 46.44万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10379292
关键词：
1-Phosphatidylinositol 3-Kinase 3-Dimensional Actins Actomyosin Acute Affect Anterior Biological Assay Biomedical Engineering Biophysics Blood Breast Cancer Cell Breast Cancer cell line Breast cancer metastasis Cancerous Cations Cell Volumes Cell membrane Cells Complement Cytoplasm Cytoskeleton Data Distant Elements Epithelial Cells Equilibrium Event Exhibits Exposure to Extracellular Matrix Extracellular Matrix Degradation Feedback Fiber Focal Adhesions Goals Growth Human body Image Imaging Device In Vitro Injections Integrins Intercellular Fluid Intuition Ion Channel Ions Light Liquid substance Lung Mechanics Mediating Memory Migration Assay Modeling Motion Motor Mucins Mus Neoplasm Metastasis Neoplasm Transplantation Nuclear Translocation Optics Organ Organoids Patients Pattern Physiological Plasma Primary Neoplasm Process Prognostic Marker Proteins Publishing Regulation Resolution Role Signal Transduction Site Speed Stretching Sum Survival Analysis Swelling Tail Testing Tissues Travel Veins Viscosity Water Zebrafish breast cancer survival cancer cell cell motility chloride-cotransporter potassium confocal imaging extracellular in vivo in vivo Model interdisciplinary approach interstitial macromolecule malignant breast neoplasm mammary mechanotransduction member migration mouse model multiphoton microscopy neoplastic cell new therapeutic target novel particle patient derived xenograft model preconditioning receptor triple-negative invasive breast carcinoma tumor

项目摘要

Cell motility is a key step in the metastatic cascade of events, as it enables cancerous cells dissociating from a primary tumor to navigate through interstitial tissues and ultimately colonize distant organs. Cell locomotion is governed by cell-matrix interactions, the actomyosin cytoskeleton, and cell volume regulation via the involvement of ion transporters, such as the Na+/H+ exchanger 1 (NHE1). To date, most cell motility assays are performed in medium with a viscosity close to that of water (0.77 cP). However, the viscosity of the interstitial fluid varies up to 2-3 cP, and can be further augmented by the presence of macromolecules secreted not only by resident epithelial cells in various tissues but also by tumor cells. Cancer cell plasticity is a key feature in metastasis, as tumor cells need to adapt to and navigate through diverse tissue microenvironments presenting different stiffness, degrees of confinement, viscosity and extracellular matrix (ECM) composition. It is currently unknown how tumor cells sense and respond to (patho)physiologically relevant levels of viscosity. The overarching goal of this project is to employ a multidisciplinary approach involving state-of-the-art bioengineering and imaging tools, quantitative analysis and in vivo models to elucidate the effects of extracellular viscosity on breast cancer cell migration, invasion and metastasis. This application will test the hypothesis, supported by intriguing preliminary data, that elevated extracellular viscosity (≥3cP) promotes NHE1-dependent cell swelling, which triggers the activation of the mechanosensitive ion channel TRPV4, thereby initiating downstream signaling. In Aim 1a, we will establish that TRPV4 is the key mechanosensor of elevated viscosity, which initiates RhoA activation, and delineate the presence of a potential feedback loop between NHE1-dependent TRPV4 activation and RhoA. In Aim 1b, we will demonstrate that the coordinated action of local isosmotic swelling at the leading edge and shrinkage at the trailing edge mediated by NHE1 and potassium-chloride cotransporter 4, KCC4, respectively, supports confined migration at elevated viscosities. Cells, as active mechanical objects upon sensing elevated extracellular viscosity, respond by balancing forces in the cell cytoplasm with those in the extracellular microenvironment, thus resulting in increased cytoskeletal tension, higher RhoA-dependent cell contractility and actin reorganization, which ultimately precipitate nuclear translocation of YAP (Aim 1c). We will characterize the roles of viscosity-sensing mechanisms in discrete steps of metastatic dissemination in a live zebrafish model that affords the unique advantages of optical transparency and exceptionally high-resolution along with high-speed imaging of transplanted tumor cells (Aim 2a). We will complement these studies with mouse models to characterize the localization patterns and functional roles of TRPV4, NHE1, KCC4 and YAP in cell migration in natural mammary tissue tracks in vivo (Aim 2b) and in breast cancer growth and metastasis (Aim 2c), using triple-negative breast cancer cell lines and patient-derived xenografts (PDXs). In sum, this project will define how cells sense and respond to extracellular viscosity and identify novel targets to reduce metastasis.

细胞运动是转移级联事件中的关键步骤，因为它使癌细胞能够从原发肿瘤解离，穿过间质组织并最终定植远处器官。细胞运动由细胞-基质相互作用、肌动球蛋白细胞骨架和细胞体积调节控制通过离子转运蛋白（例如 Na+/H+ 交换器 1 (NHE1)）的参与，迄今为止，大多数细胞的运动性都受到影响。测定是在粘度接近水 (0.77 cP) 的介质中进行的。间质液变化高达 2-3 cP，并且可以因分泌的大分子的存在而进一步增加不仅通过各种组织中的常驻上皮细胞，而且通过肿瘤细胞的可塑性是一个关键特征。在转移中，由于肿瘤细胞需要适应并穿越不同的组织微环境目前，不同的硬度、限制程度、粘度和细胞外基质（ECM）成分。未知肿瘤细胞如何感知和响应（病理）生理相关的粘度水平。该项目的总体目标是采用涉及最先进生物工程的多学科方法和成像工具、定量分析和体内模型来阐明细胞外粘度对乳腺癌细胞迁移、侵袭和转移该应用将检验该假设，并得到支持。有趣的初步数据表明，细胞外粘度升高 (≥3cP) 会促进 NHE1 依赖性细胞肿胀，触发机械敏感离子通道 TRPV4 的激活，从而启动下游在目标 1a 中，我们将确定 TRPV4 是启动粘度升高的关键机械传感器。 RhoA 激活，并描绘了 NHE1 依赖性 TRPV4 之间潜在反馈回路的存在在目标 1b 中，我们将证明局部等渗肿胀的协调作用。由 NHE1 和氯化钾协同转运蛋白 4 介导的前缘和后缘收缩， KCC4 分别支持细胞在高粘度下的有限迁移，作为活动的机械物体。感知细胞外粘度升高，通过平衡细胞质中的力与细胞中的力来做出反应细胞外微环境，从而导致细胞骨架张力增加，RhoA依赖性细胞更高收缩性和肌动蛋白重组，最终促进 YAP 的核易位（目标 1c）。表征粘度传感机制在活体内转移传播的离散步骤中的作用斑马鱼模型具有光学透明和高分辨率的独特优势以及移植肿瘤细胞的高速成像（目标 2a）。小鼠模型来表征 TRPV4、NHE1、KCC4 和 YAP 的定位模式和功能作用天然乳腺组织中的细胞迁移跟踪体内（目标 2b）以及乳腺癌生长和转移（目标 2c），使用三阴性乳腺癌细胞系和患者来源的异种移植物（PDX）总之，该项目。将定义细胞如何感知和响应细胞外粘度，并确定减少转移的新靶点。