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：
10559616
负责人：
Konstantinos Konstantopoulos
金额：
$ 48.21万
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/10559616
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 Separation Cell Volumes Cell membrane Cell surface Cells Cellular biology Cessation of life Clinical Complement Confined Spaces Cytoplasm Cytoskeleton Data Dissociation 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 Mediating Metastatic breast cancer Microfluidic Microchips Microtubules Modeling Molecular Molecular Biology Mus Myosin Type II Neoplasm Metastasis Optics Organ Organoids Osmosis Patients Phase Primary Neoplasm Regulation Research Personnel Role Solid Neoplasm Swelling Testing Tissues Traction 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 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.

临床上无法治疗肿瘤转移是导致大多数癌症患者的原因细胞迁移是癌细胞从原发肿瘤转移到扩散的关键步骤。体内远端器官的细胞运动受细胞基质相互作用、肌动球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，通过渗透引擎模型 (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的方法。方法涉及最先进的生物工程和成像工具、定量分析和体内模型定义乳腺癌中离子转运蛋白和细胞骨架之间的相对作用和潜在串扰该应用将测试这一假设，并得到有趣的初步支持。数据表明，前缘局部等渗膨胀和后缘收缩的协调作用分别由 NHE1 和 SWELL1 介导，支持限制内的迁移。 NHE1 和 SWELL1 与细胞骨架协同作用，介导有效的迁移和转移。由于缺乏针对三阴性乳腺癌（TNBC）的靶向治疗，我们将优先考虑 TNBC 细胞系和患者在目标 1 中，我们将建立 NHE1 和 NHE1 的功能作用。 SWELL1 在不同刚度的限制 µ 通道内的细胞迁移、3D 凝胶和细胞传播中的作用我们还将阐明导致极化的机制。 NHE1 和 SWELL1 分别在细胞前部和后部的分布，并使用新颖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。与此同时，我们将开发一个创新的数学模型来识别关键变量，使 OEM- 在目标 2 中，我们将描述 OEM 和各种细胞骨架之间的相互作用。成分，包括 b1 整合素、肌球蛋白 II、肌动蛋白和微管。重要的是，我们将定义细胞内。负责 NHE1 和 SWELL1 沿着细胞纵向表面穿梭的运输机制。还引入了一个全面的数学模型来破译 OEM 和细胞骨架的串扰在目标 3 中，我们将展示 NHE1 和 SWELL1 的作用。天然乳腺组织中细胞迁移的沉默可在体内追踪并检查它们在乳腺癌中的作用使用原位移植到第四乳腺脂肪垫的 TNBC 细胞系和 PDX 进行生长和转移我们将通过斑马鱼实验来补充小鼠研究，这使我们能够对其整体进行成像。以卓越的光学清晰度观察脉管系统，以描绘离子转运蛋白在不同步骤中的作用该应用程序汇集了具有生物工程专业知识的研究人员团队，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。