Quantitative analysis of epithelial cell scatter

上皮细胞分散的定量分析

• 批准号: 8027766
• 负责人: ANAND R ASTHAGIRI
• 金额: $ 26.78万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-11 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8027766
• 关键词: Quantitative; analysis; epithelial; cell; scatter

DESCRIPTION (provided by applicant): The major aim of the proposed work is to develop a quantitative understanding of epithelial cell scatter, a process that is closely linked to late metastatic stages of cancer development. During metastasis, epithelial tissue structure is significantly disrupted as epithelial cells escape from their primary site and invade surrounding tissue. An effective ex vivo model of metastasis involves cell scatter. Epithelial cells grow in clusters ex vivo, reminiscent of their monolayer, well-packed morphology in vivo. Metastasis-associated genetic perturbations promote cells to "peel away" from clusters and scatter into the surrounding region. Thus, the cell scatter assay has been used to identify oncogenes (OGs) and tumor suppressor genes (TSGs) that may play a role in metastasis. However, current approaches are limited to qualitative characterizations from which it is difficult to assess how potent a particular OG/TSG might be and which combinations exhibit the most synergism. Answers to such questions can guide us to the most potent choice of drug targets and could help us design the most effective combination treatments. Furthermore, to understand more deeply how OG/TSGs quantitatively affect population-level phenotype, it is essential to examine the cell-level processes that contribute to multicellular scatter. These cell-level properties include the migration of individual cells and multicellular groups. In addition, migrating cells will collide. Whether these collisions re-seed small clusters or whether colliding cells "bounce apart" (akin to an elastic collision) will affect the extent and dynamics of cell scatter. Current techniques to quantify these cell-level properties are too cumbersome to permit quantitative, systems-scale analysis of the effects of numerous OG/TSGs. In the proposed work, we seek to address these challenges in order to elucidate the quantitative effects of the adhesive microenvironment and OG/TSGs on cell scatter. We will quantify both the cell- and population-level aspects of cell scatter using a multi-faceted strategy that integrates automated high-throughput imaging and micropatterning. The Specific Aims are: 1. To develop quantitative automated methods for measuring cell-level motility properties. 2. To elucidate the quantitative effect of the adhesive microenvironment on multicellular scatter and the underlying cell-level properties. 3. To elucidate the quantitative contributions and synergisms of metastatic genes to multicellular scatter and to the underlying cell-level properties. Results from the proposed work will provide a deeper quantitative understanding of how OG/TSGs and the adhesive microenvironment affect cell-level motile properties and multicellular scatter, a process closely related to metastasis. PUBLIC HEALTH RELEVANCE: The majority of human cancers occurs in epithelial tissues, and the disease enters a lethal metastatic phase when cancer cells escape from well-ordered epithelial tissues and infiltrate into surrounding areas. The proposed work will provide a deeper quantitative understanding of how genetic changes induce cells to "scatter" from their neighbors, a core aspect of metastasis. These quantitative insights will point us to the most potent scatter-inducing genes, highlighting potentially more effective drug targets to curb metastatic processes.

描述（由申请人提供）：拟议工作的主要目的是发展上皮细胞分散的定量理解，这是一个与癌症发展的晚期转移阶段密切相关的过程。在转移过程中，上皮组织结构被显著破坏，因为上皮细胞从其原发部位逃逸并侵入周围组织。转移的有效离体模型涉及细胞分散。上皮细胞在体外成簇生长，使人联想到它们在体内的单层、包装良好的形态。转移相关的遗传扰动促进细胞从簇中“剥离”并分散到周围区域。因此，细胞散射测定已被用于鉴定可能在转移中起作用的癌基因（OG）和肿瘤抑制基因（TSG）。然而，目前的方法仅限于定性表征，难以评估特定OG/TSG的效力以及哪些组合表现出最大的协同作用。这些问题的答案可以指导我们选择最有效的药物靶点，并帮助我们设计最有效的联合治疗。此外，为了更深入地了解OG/TSGs如何定量地影响群体水平的表型，必须检查有助于多细胞分散的细胞水平过程。这些细胞水平的特性包括单个细胞和多细胞群体的迁移。此外，迁移的细胞会发生碰撞。这些碰撞是否重新播种小簇或碰撞细胞是否“弹开”（类似于弹性碰撞）将影响细胞分散的程度和动力学。目前的技术来量化这些细胞水平的属性是太麻烦，允许定量，系统规模的分析，许多OG/TSG的影响。 在拟议的工作中，我们试图解决这些挑战，以阐明细胞分散的粘附微环境和OG/TSGs的定量影响。我们将使用一种多方面的策略来量化细胞散射的细胞和群体水平方面，该策略集成了自动化高通量成像和微图案化。具体目标是：1。建立细胞水平运动特性的自动化定量测定方法。2.阐明粘附微环境对多细胞散射的定量影响和潜在的细胞水平特性。3.阐明转移基因对多细胞分散和细胞水平特性的定量贡献和协同作用。从拟议的工作的结果将提供一个更深入的定量了解OG/TSGs和粘附微环境如何影响细胞水平的运动特性和多细胞散射，一个过程密切相关的转移。 公共卫生关系：大多数人类癌症发生在上皮组织中，当癌细胞从有序的上皮组织中逃逸并浸润到周围区域时，疾病进入致命的转移阶段。拟议的工作将提供一个更深入的定量了解如何遗传变化诱导细胞从他们的邻居“散射”，转移的核心方面。这些定量的见解将为我们指出最有效的散射诱导基因，突出潜在的更有效的药物靶点，以遏制转移过程。