The Role of Physical Cues in Collective Cell Invasion

物理线索在集体细胞入侵中的作用

基本信息

批准号：
10016201
负责人：
Konstantinos Konstantopoulos
金额：
$ 31.3万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-29 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10016201
关键词：
3-Dimensional Actomyosin Adhesions Anatomy Architecture Automobile Driving Basement membrane Biological Assay Biomedical Engineering Blood Cell Adhesion Cell Culture System Cell Culture Techniques Cell Line Cell Shape Cell model Cell-Cell Adhesion Cells Cellular biology Characteristics Chemicals Collagen Complement Complex Computer Models Connective Tissue Cues Data Development Distant Distant Metastasis E-Cadherin EGF gene Epithelial Epithelium Event Experimental Models Extracellular Matrix Frequencies Genes Genetically Engineered Mouse Geometry Imaging Techniques In Vitro Individual Invaded Light Locomotion Macrophage Colony-Stimulating Factor Malignant Neoplasms Mammary Neoplasms Mechanics Microfluidic Microchips Microfluidics Modeling Molecular Monomeric GTP-Binding Proteins Mus Muscle Neoplasm Metastasis Nerve Oncology Optics Organ Pathologist Primary Neoplasm Process Prognostic Marker Recurrence Role Series Signal Transduction Site Stream System Testing Tissues Transplantation Tumor Cell Invasion Width Work base cancer cell cancer subtypes cell motility cellular imaging comparative epithelial to mesenchymal transition experimental study genetic signature in vivo in vivo Model innovation insight interdisciplinary approach intravital imaging intravital microscopy leukemia/lymphoma loss of function malignant breast neoplasm mathematical model microdevice migration mouse model multiphoton imaging neoplastic cell novel physical property physical science real-time images response three dimensional cell culture tool tumor

项目摘要

Summary of Project 1: The Role of Physical Cues in Collective Cell Invasion The ability of tumors to invade adjacent tissues, leading to local or distant metastasis, is a hallmark of cancer. Cancer cells frequently invade as groups of adherent cells in a process termed collective invasion. Previous studies have primarily focused on single cell or semi-collective (multicellular streaming) cell invasion. Single cell models for metastasis have direct implications for tumors whose cells migrate constitutively as individual cells, such as leukemias and lymphomas, or after cell detachment from a primary tumor via epithelial-to- mesenchymal transition (EMT). However, EMT has long been controversial among pathologists as breast tumors at metastatic sites typically display epithelial features. While EMT-like gene signatures can be observed in specific mouse models and breast cancer subtypes, the majority of breast tumors do not exhibit clear molecular features of EMT. Intravital microscopy studies reveal that tumor cells preferentially migrate collectively along pre-existing channels that are defined by various anatomical structures in vivo. However, it is currently unknown how the physical properties of the microenvironment, such as confinement and compliance, regulate the molecular mechanisms of collective cell invasion. Intriguing preliminary data reveal that cancer cells migrate through wide (≥50 µm) tracks as a collective unit. However, as confinement increases, the cancer cells spontaneously disseminate, first as clusters of 2-5 cells and eventually, in very narrow tracks (≤10 µm), as single cells. We hypothesize that the physical microenvironment induces a signaling cascade of events that transforms the classical collective to single cell invasion. To test this hypothesis, we will employ a multidisciplinary approach combining novel bioengineering tools and mathematical modeling with sophisticated molecular cell biology and imaging techniques and in vivo models. In Aim 1, we will develop an integrated experimental and computational model of collective cell movement in confined geometries modeling primary tumor invasion, and dissect the mechanisms by which cell-cell contact is released during mechanically-induced transitions to single cell movement, focusing on the role of E-cadherin cleavage and possible EMT induction. In Aim 2, we will delineate the relative contributions of actomyosin contractility, small GTPases and osmotic engine model to locomotion in rigid versus compliant confined microenvironments. In Aim 3, we will validate our in vitro understanding of the dissemination and locomotion of cancer cells in more complex microenvironments characteristic of in vivo breast tumors using an organotypic 3D culture system and genetically engineered mouse models. Elucidation of the underlying mechanisms of collective cancer cell invasion will offer insights into our understanding of how cancer cells spread through the body, and it could shift the currently prevailing single cell paradigm in cancer to incorporate concepts of mechanical signaling, cell-cell adhesion, and cell-cell cooperation.

项目1的摘要：物理线索在集体细胞入侵中的作用肿瘤侵入相邻组织（导致局部或远处转移）的能力是癌症的标志。在称为集体侵袭的过程中，癌细胞经常侵入作为依从细胞的组。以前的研究主要集中于单细胞或半体体（多细胞流）侵袭。单身的转移的细胞模型对肿瘤的细胞作为个体迁移的肿瘤具有直接影响细胞，例如白血病和淋巴瘤，或通过上皮到原发性肿瘤的细胞脱离后的细胞间充质转变（EMT）。但是，EMT长期以来一直在病理学家作为乳房引起争议转移部位的肿瘤通常显示上皮特征。虽然可以观察到EMT样基因特征在特定的小鼠模型和乳腺癌亚型中，大多数乳腺肿瘤不暴露明显 EMT的分子特征。弹性显微镜研究表明，肿瘤细胞优先迁移沿着体内各种解剖结构定义的先前的通道统称。但是，是目前未知微环境的物理特性，例如限制和合规性，调节集体细胞侵袭的分子机制。有趣的初步数据表明癌症细胞通过宽（≥50µm）的轨道迁移，作为集体单位。但是，随着监禁的增加，癌症细胞赞助传播，首先是2-5个细胞的簇，最终以非常狭窄的轨道（≤10µm）为单细胞。我们假设物理微环境引起了一系列信号的事件，这些事件是将经典集体转变为单细胞入侵。为了检验这一假设，我们将采用多学科方法将新颖的生物工程工具和数学建模与精致的分子细胞生物学和成像技术以及体内模型。在AIM 1中，我们将开发一个集成的限制几何形状建模主要的集体细胞运动的实验和计算模型肿瘤侵袭，并剖析机械诱导期间释放细胞细胞接触的机制向单细胞运动的过渡，重点是E-钙粘蛋白裂解和可能的EMT诱导的作用。在 AIM 2，我们将描述Actomyosin收缩力，小GTPases和渗透的相对贡献发动机模型，以刚性与符合统一的限制微环境。在AIM 3中，我们将验证我们对更复杂的癌细胞的传播和运动的体外理解使用有机3D培养系统和基因工程的鼠标模型。阐明集体癌细胞的基本机制入侵将为我们了解我们对癌细胞如何通过身体传播的理解，并且可以将当前主要的单细胞范式转移到癌症中，以结合机械信号的概念，细胞细胞粘附和细胞 - 细胞合作。