Transcriptional control of collective cell migration

集体细胞迁移的转录控制

• 批准号: 8725189
• 负责人: Lionel Christiaen
• 金额: $ 29.41万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-23 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8725189
• 关键词: Address; Animals; Attention; Automobile Driving; Binding; Biological Assay; Biological Models; Bone Morphogenetic Proteins; Cardiac; Cell Polarity; Cell physiology; Cells; Chimeric Proteins; Chordata; Ciona intestinalis; Competence; Complement; Development; Dominant-Negative Mutation; Embryo; Embryonic Development; Event; Fibroblast Growth Factor; Fluorescence Resonance Energy Transfer; Gene Targeting; Genes; Genetic Transcription; Genome; Goals; Guanosine; Human; Image; In Situ Hybridization; Life; Marine Invertebrates; Membrane; Mesoderm; Mesoderm Cell; Microarray Analysis; Microfilaments; Modeling; Molecular; Neoplasm Metastasis; Output; Pathologic Processes; Physiological; Physiological Processes; Prevention; Process; Regenerative Medicine; Regulation; Regulator Genes; Regulatory Element; Research; Side; Signal Transduction; Source; Structure; System; Testing; Tissues; Transcriptional Regulation; Urochordata; Vertebrates; Work; Wound Healing; angiogenesis; ascidian; base; cell behavior; cell motility; expectation; imaging modality; insight; membrane polarity; migration; rho; rho GTP-Binding Proteins; transcription factor

PROJECT SUMMARY / ABSTRACT Critical physiological and pathological processes, such as wound healing, de novo vessel angiogenesis and cancer metastasis, rely on directed collective cell migrations, whereby groups of cells become polarized and moves together in an orderly fashion. The ability of cell groups to migrate collectively is determined in part by the tissue-specific transcriptional inputs that define the complement of genes that they express and thus their competence to migrate. The long-term goal of this project is to understand how tissue-specific transcription regulators control the basic cellular processes underlying directed collective cell migration. To this aim, the simplified embryos of a chordate species, the ascidian Ciona intestinalis, will be used to study the migration of pre-cardiac mesoderm cells, called "trunk ventral cells" (TVCs). The TVCs provide the simplest possible model of directed collective cell migration in live embryos. On each side of the embryo, only two cells migrate together and display a clear Leader-Trailer (L-T) polarity aligned with the direction of migration: the leader TVC displays a broad leading edge and more conspicuous protrusions than the trailer. It was previously established that Mesp, Fibroblast Growth Factor (FGF) signaling and FoxF transcriptional inputs determine the ability of TVCs to migrate. The specific goal of the proposed research is to understand how transcriptional inputs contribute to the specification of the leader TVC and influence its protrusive activity. On the basis of preliminary observations, the hypotheses that (1) a ventral source of bone morphogenetic proteins (BMP) contributes to the L-T polarized expression of downstream target genes, including the Lef/Tcf transcription factor; and (2) these transcriptional inputs determine the L-T polarized activities of the Rho GTPases RhoDF and Cdc42, which are required for the formation of membrane protrusions will be tested. To address these possibilities, the expression, cis-regulation and function of Lef/Tcf in the TVCs will first be analyzed in order to understand L-T polarized transcriptional inputs. Second, quantitative imaging methods will be used to document the L-T polarization of RhoDF, Cdc42 and protrusive activities in wild-type conditions and after manipulations of transcription regulators and other candidate Rho GTPases in an attempt to characterize their effects on L-T polarized membrane protrusions. Finally, candidate regulated effectors of collective cell polarization and protrusive activity will be identified using TVC-specific whole genome microarray analysis followed by systematic expression and functional analyses by in situ hybridization and targeted over-expression, respectively. The expectation is to determine a mechanism of transcriptional control for collective cell migration by characterizing the transcription regulators, the regulated effectors and their effects on the Rho GTPase signaling and actin filament dynamics that underlie collective cell polarization and the formation of membrane protrusions.

项目总结/摘要 关键的生理和病理过程，如伤口愈合、新生血管生成和 癌症转移依赖于定向的集体细胞迁移，由此细胞群变得极化， 有序地移动细胞群集体迁移的能力部分取决于 组织特异性转录输入，其定义了它们表达的基因的互补，因此它们的 移民的能力。这个项目的长期目标是了解组织特异性转录是如何 调节因子控制着作为定向集体细胞迁移基础的基本细胞过程。为此， 一个脊索动物物种的简化胚胎，海鞘玻璃海鞘，将被用来研究迁移， 心前中胚层细胞，称为“躯干腹侧细胞”（TVC）。TVC提供了最简单的模型 在活的胚胎中定向的集体细胞迁移。在胚胎的每一侧，只有两个细胞一起迁移 并显示与迁移方向对齐的清晰的引导-尾随（L-T）极性：引导TVC显示 宽的前缘和比拖车更明显的突起。此前已确定， Mesp、成纤维细胞生长因子（FGF）信号传导和FoxF转录输入决定TVC的能力， 迁徙拟议研究的具体目标是了解转录输入如何有助于 领导者TVC的规格，并影响其扩张活动。根据初步 观察，假设（1）骨形态发生蛋白（BMP）的腹侧来源有助于 下游靶基因的L-T极化表达，包括Lef/Tcf转录因子;和（2） 这些转录输入决定RhoGTP酶RhoDF和Cdc 42的L-T极化活性， 将测试形成膜突起所需的量。为了解决这些可能性， 为了了解L-T，首先分析TVC中Lef/Tcf的表达、顺式调节和功能， 极化的转录输入。其次，定量成像方法将用于记录L-T RhoDF、Cdc 42的极化和在野生型条件下和在操作后的抑制活性。 转录调节因子和其他候选Rho GTP酶，试图表征它们对L-T 极化膜突起。最后，候选调节效应集体细胞极化和 将使用TVC特异性全基因组微阵列分析，随后使用 通过原位杂交和靶向过表达进行系统表达和功能分析， 分别期望确定集体细胞迁移的转录控制机制 通过表征转录调节因子、受调节的效应因子及其对Rho GT3的影响， 信号传导和肌动蛋白丝动力学是集体细胞极化和膜形成的基础 突起。