The interplay of ion transporters and cytoskeleton in breast cancer migration and metastasis

离子转运蛋白和细胞骨架在乳腺癌迁移和转移中的相互作用

基本信息

批准号：
10338164
负责人：
Konstantinos Konstantopoulos
金额：
$ 48.73万
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：
2021-02-02 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10338164
关键词：
3-Dimensional Actins Actomyosin Affect Automobile Driving Biological Models Biomedical Engineering Breast Cancer Cell Breast Cancer cell line Breast cancer metastasis Cancer Biology Cancer Patient Cancerous Cell Cycle Cell Volumes Cell membrane Cell surface Cells Cellular biology Cessation of life Clinical Complement Confined Spaces Cytoplasm Cytoskeleton Data Devices Distant Elements Event Extracellular Matrix Fatty acid glycerol esters Fiber Gel Goals Growth Image Imaging Device In Vitro Individual Integrins Intracellular Transport Ion Channel Ions Light Liquid substance Mediating Metastatic breast cancer Microtubules Modeling Molecular Molecular Biology Mus Myosin Type II Neoplasm Metastasis Optics Organ Organoids Patients Phase Primary Neoplasm Regulation Research Personnel Role Solid Neoplasm Swelling Testing Tissues Transplantation Travel Tumor Cell Migration Water Work Zebrafish cancer cell cell motility combat experimental study fluid flow in vivo in vivo Model innovation interdisciplinary approach interstitial intravital microscopy live cell imaging malignant breast neoplasm mammary mathematical model migration mouse model multiphoton microscopy neoplastic cell new therapeutic target novel optogenetics patient derived xenograft model targeted treatment therapeutic target tool triple-negative invasive breast carcinoma tumor tumor xenograft tumorigenic water channel

项目摘要

The inability to clinically treat tumor metastasis is responsible for the majority of cancer patient deaths. Cell migration is a pivotal step in the metastatic dissemination of cancer cells from a primary tumor to distant organs in the body. Cell motility 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), as explained by the Osmotic Engine Model (OEM). The roles of cytoskeleton and ion transporters in cell locomotion have been typically studied in isolation. 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 define the relative roles and potential crosstalk between ion transporters and the cytoskeleton in breast cancer cell migration and metastasis in vivo. This application will test the hypothesis, supported by intriguing preliminary data, that the coordinated action of local isosmotic swelling at the leading edge and shrinkage at the trailing edge mediated by NHE1 and SWELL1, respectively, supports migration in confinement. We further hypothesize that NHE1 and SWELL1 act in concert with cell cytoskeleton to mediate efficient migration and metastasis. Given the lack of targeted therapies for triple negative breast cancer (TNBC), we will prioritize TNBC cell lines and patient- derived xenograft (PDX) tumor cells as models. In Aim 1, we will establish the functional roles of NHE1 and SWELL1 in cell migration inside confining µ-channels of different stiffnesses, in 3D gels and in cell dissemination from 3D breast cancer cell organoids. We will also elucidate the mechanism responsible for the polarized distribution of NHE1 and SWELL1 at the cell front and rear, respectively, and use novel optogenetic tools to alter their spatial polarization and test how these alterations affect the direction and efficiency of cell migration. In parallel, we will develop an innovative mathematical model to identify the key variables that enable OEM- mediated cell motility. In Aim 2, we will delineate the interplay between OEM and the various cytoskeletal constituents, including b1 integrins, myosin II, actin and microtubules. Importantly, we will define the intracellular transport mechanisms responsible for NHE1 and SWELL1 shuttling along the longitudinal cell surface. We will also introduce a comprehensive mathematical model to decipher the crosstalk of OEM and cytoskeletal components in regulating migration efficiency. In Aim 3, we will demonstrate the effects of NHE1 and SWELL1 silencing on cell migration in natural mammary tissue tracks in vivo and examine their roles in breast cancer growth and metastasis, using TNBC cell lines and PDXs orthotopically transplanted to the 4th mammary fat pad of mice. We will complement mouse studies with experiments in zebrafish, which enables us to image its entire vasculature at exceptional optical clarity, in order to delineate the roles of ion transporters in different steps of the metastatic cascade. This application brings together a team of investigators with expertise in bioengineering, imaging, cell & molecular biology, quantitative analysis, PDXs, in vivo studies and breast cancer biology.

无法临床治疗肿瘤转移是癌症患者的大多数死亡人数。细胞迁移是癌细胞从原发性肿瘤转移到转移性的关键步骤体内遥远的器官。细胞运动性受细胞玛尔体相互作用，肌动蛋白细胞骨架和通过离子转运蛋白（例如Na+/H+交换器1（NHE1））的参与来调节细胞体积由渗透引擎模型（OEM）解释。细胞骨架和离子转运蛋白在细胞运动中的作用通常是孤立研究的。该项目的总体目标是采用多学科涉及最先进的生物工程和成像工具，定量分析以及体内模型的方法定义离子转运蛋白和乳腺癌细胞骨架之间的相对作用和潜在的串扰细胞迁移和体内转移。该应用程序将检验该假设，并通过引人入胜的初步支持数据，局部同质肿胀在前沿的协调作用，在后缘收缩由NHE1和Swell1介导，分别支持完成迁移。我们进一步假设 NHE1和SWELL1与细胞骨骼协同作用，以介导有效的迁移和转移。鉴于缺乏针对三重阴性乳腺癌（TNBC）的靶向疗法，我们将优先考虑TNBC细胞系和患者 - 衍生的异种移植（PDX）肿瘤细胞作为模型。在AIM 1中，我们将确定NHE1和在限制不同刚度，3D凝胶和细胞传播中的细胞迁移中的swell1迁移。来自3D乳腺癌细胞器官。我们还将阐明负责极化的机制 NHE1和Swell1在细胞前后分别分布，并使用新型的光遗传学工具来改变它们的空间极化并测试这些改变如何影响细胞迁移的方向和效率。在并行，我们将开发一个创新的数学模型，以确定使OEM-的关键变量介导的细胞运动。在AIM 2中，我们将描述OEM与各种细胞骨架之间的相互作用包括B1整联蛋白，肌球蛋白II，肌动蛋白和微管包括的组成量。重要的是，我们将定义细胞内负责NHE1和Swell1沿纵向细胞表面穿梭的运输机制。我们将还引入了一个全面的数学模型，以解读OEM和细胞骨架的串扰控制迁移效率的组成部分。在AIM 3中，我们将演示NHE1和Swell1的影响在体内沉默的天然乳腺组织轨迹中的细胞迁移并检查其在乳腺癌中的作用生长和转移，使用TNBC细胞系和原位移植到第四乳腺脂肪垫的PDXS 小鼠。我们将通过斑马鱼中的实验补充小鼠研究，这使我们能够图像整个图像在特殊光学清晰度下的脉管系统，以在不同的步骤中描述离子转运蛋白的作用转移性级联。该应用程序汇集了一组研究人员，具有生物工程专业知识，成像，细胞和分子生物学，定量分析，PDX，体内研究和乳腺癌生物学。