Transcriptional control of collective cell migration

集体细胞迁移的转录控制

基本信息

批准号：
9344648
负责人：
Lionel Christiaen
金额：
$ 31.96万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-23 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9344648
关键词：
Actins Adhesions Alpha Cell Animals Attention Automobile Driving Basal lamina Behavior Biological Assay CRISPR/Cas technology Cardiac Cell Polarity Cell membrane Cell physiology Cell-Matrix Junction Cells Chordata Ciona intestinalis Collagen Competence Complement Computer Simulation Development Embryo Embryonic Development Endocytosis Endoderm Endosomes Epidermis Extracellular Matrix Fibroblast Growth Factor Future Gene Expression Genes Genetic Transcription Germ Layers Goals Integrins Invertebrates Libraries Link MAP Kinase Gene Maintenance Maps Mediating Mesoderm Cell Modeling Monitor Neoplasm Metastasis Output Pathologic Pathologic Processes Pattern Phenotype Physiological Physiological Processes Positioning Attribute Process Property Receptor Protein-Tyrosine Kinases Receptor Signaling Recycling Regenerative Medicine Regulation Regulator Genes Research Role Side Signal Transduction Structure System Talin Testing Tissues Transcriptional Regulation Urochordata Vascular Endothelial Growth Factors Vertebrates Vesicle Wound Healing ascidian blood vessel development cancer prevention cell behavior cell motility discoidin domain receptor 2 discoidin receptor imaging modality insight interest loss of function metastasis prevention migration novel polarized cell progenitor quantitative imaging receptor receptor expression receptor function receptor upregulation rho GTP-Binding Proteins trafficking transcriptome sequencing

项目摘要

PROJECT SUMMARY / ABSTRACT Critical physiological and pathological processes, such as wound healing, blood vessel formation and cancer metastasis, rely on directed collective cell migrations, whereby groups of cells collectively polarize and move together in an orderly fashion. The ability of cell collectives to migrate directionally is determined in part by the tissue-specific transcriptional inputs that define the complement of expressed genes 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 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. Moreover, TVCs migrate strictly between the endodermal and ectodermal germ layers, a hallmark of mesodermal cardiac progenitors. It was determine that these surrounding tissues contribute to canalizing TVCs' innate motility towards collective polarity and directed migration. The goal of the proposed research is to understand how transcriptionally-controlled intrinsic TVC properties interface with extrinsic signals to determine collective polarity and directed migration in the embryo. Preliminary studies suggested that the gene encoding the discoidin domain receptor (Ddr) is upregulated by Mesp, FGF and FoxF transcriptional inputs in the TVCs, where it promotes adhesion to the epidermis. Using newly developed quantitative imaging methods, the detailed mechanisms controlling Ddr expression, localization and activity will be analyzed. The hypothesis that a cell-autonomous antagonism between Ddr and vascular endothelial growth factor receptor (Vegfr) signaling positions the migrating TVCs between the epidermis and endoderm will be tested. Preliminary observations suggest that Ddr promotes adhesion to the epidermis by regulating vesicle trafficking. The hypothesis that Ddr acts in Rab4/Rab11-positive endosomes to promote the recycling of integrins to the plasma membrane will be tested. Finally, the functions of regulated candidate effectors of collective migration will be studied extensively using TVC-specific CRISPR/Cas9-mediated loss-of-function assays and high-content phenotypic analyses. A provisional model of the biomolecular network controlling the subcellular processes underlying TVC behavior will be built. Particular attention will be paid to the candidate modulators of Ddr, Vegfr and integrin functions. Completion of this project will illuminate the systems' level mechanisms linking intrinsic transcriptional inputs and extrinsic signals to define cell-specific behaviors.

项目摘要 /摘要关键的生理和病理过程，例如伤口愈合，血管形成和癌症转移，依赖于定向的集体细胞迁移，在该迁移中，细胞组集体极化和移动以有序的方式在一起。细胞集体在方向迁移的能力部分由组织特异性的转录输入，定义了表达基因的完成及其能力迁移。该项目的长期目标是了解组织特定的转录调节器控制定向集体细胞迁移基础的基本细胞过程。为此，简化了脊柱物种的胚胎，海腹肠道肠将用于研究前的迁移心脏中胚层细胞，称为“躯干腹细胞”（TVC）。 TVC提供了最简单的模型在活胚胎中定向集体细胞迁移。在胚胎的每一侧，只有两个细胞一起迁移并显示一个清晰的领导者拖车极性与移民方向一致：领导者TVC显示A 宽阔的前缘和比拖车更明显的蛋白质。以前已经确定Mesp，成纤维细胞生长因子（FGF）信号传导和FOXF转录输入确定TVC的能力迁移。此外，TVC严格迁移到内胚层和外胚层细菌层，这是一个标志中胚层心脏祖细胞。已经确定这些周围的组织有助于引导 TVCS天生的运动能力朝着集体极性和定向移民。拟议研究的目的是了解转录控制的固有TVC属性如何与外部信号接口到确定胚胎中的集体极性和定向迁移。初步研究表明该基因 MESP，FGF和FOXF转录输入在编码盘状蛋白域受体（DDR）中进行编码 TVC，它促进了对表皮的依从性。使用新开发的定量成像方法，将分析控制DDR表达，定位和活性的详细机制。假设 DDR和血管内皮生长因子受体（VEGFR）之间的细胞自主拮抗作用信号定位将测试表皮和内胚层之间的迁移TVC。初步的观察结果表明，DDR通过调节囊泡贩运来促进表皮依从性。这 DDR作用于Rab4/Rab11阳性内体以促进整联蛋白对等离子体的回收的假设膜将进行测试。最后，受监管的集体迁移候选效应的功能将是使用TVC特异性CRISPR/CAS9介导的功能丧失测定和高含量进行广泛研究表型分析。控制亚细胞过程的生物分子网络的临时模型将建立基本的TVC行为。特别关注DDR的候选调制器， VEGFR和整联蛋白功能。该项目的完成将阐明系统的级别机制内在的转录输入和外部信号，以定义细胞特异性行为。