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 总结：物理线索在集体细胞入侵中的作用肿瘤侵入邻近组织、导致局部或远处转移的能力是癌症的标志。癌细胞经常作为贴壁细胞群进行侵袭，这一过程称为集体侵袭。研究主要集中在单细胞或半集体（多细胞流）单细胞入侵。转移的细胞模型对细胞作为个体进行组成型迁移的肿瘤有直接影响细胞，例如白血病和淋巴瘤，或细胞通过上皮细胞从原发性肿瘤脱离后间质转化（EMT）然而，EMT 长期以来一直在病理学家中与乳腺疾病一样存在争议。转移部位的肿瘤通常表现出上皮特征，同时可以观察到类似 EMT 的基因特征。在特定的小鼠模型和乳腺癌亚型中，大多数乳腺肿瘤没有表现出明显的 EMT 的分子特征。活体显微镜研究表明肿瘤细胞优先迁移。然而，它是沿着由体内各种解剖结构定义的预先存在的通道共同形成的。目前尚不清楚微环境的物理特性（例如限制和顺应性）如何调节集体细胞侵袭的分子机制有趣的初步数据揭示了癌症。细胞作为一个集体单元通过宽（≥50 µm）的轨道迁移，但是，随着限制的增加，癌症。细胞自发传播，首先以 2-5 个细胞的簇形式传播，最终以非常窄的轨迹 (≤10 µm) 传播，如我们发现物理微环境会引发一系列信号级联事件，将经典的集体入侵转变为单细胞入侵。多学科方法将新颖的生物工程工具和数学建模与复杂的在目标 1 中，我们将开发一种集成的分子细胞生物学和成像技术以及体内模型。有限几何中集体细胞运动的实验和计算模型模拟初级肿瘤侵袭，并剖析机械诱导过程中细胞与细胞接触释放的机制过渡到单细胞运动，重点关注 E-钙粘蛋白裂解和可能的 EMT 诱导的作用。目标 2，我们将描述肌动球蛋白收缩性、小 GTP 酶和渗透压的相对贡献在目标 3 中，我们将验证发动机模型在刚性与柔性受限微环境中的运动。我们对癌细胞在更复杂的情况下传播和运动的体外理解使用器官型 3D 培养系统研究体内乳腺肿瘤的微环境特征基因工程小鼠模型阐明集体癌细胞的潜在机制。入侵将使我们深入了解癌细胞如何在体内传播，并且它可以改变目前流行的癌症单细胞范例，纳入机械信号传导的概念，细胞与细胞的粘附和细胞与细胞的合作。