Elucidating the mechanisms underlying cell cycle regulation of invasive behavior

阐明侵袭行为的细胞周期调控机制

• 批准号: 9919976
• 负责人: David Matus
• 金额: $ 2.58万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-13 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9919976
• 关键词: Adopted; Alleles; Asthma; Basement membrane; Behavior; Biological Assay; Caenorhabditis elegans; Cardiac development; Cell Cycle; Cell Cycle Arrest; Cell Cycle Regulation; Cell division; Cells; Chromatin; Chromatin Remodeling Factor; Clustered Regularly Interspaced Short Palindromic Repeats; Cytoskeleton; Data; Defect; Development; Disease; Disease Progression; Disseminated Malignant Neoplasm; Embryonic Development; Epigenetic Process; Epithelium; Euchromatin; F-Actin; Failure; G1 Arrest; G1 Phase; Gene Expression; Genes; Genetic; Genetic Diseases; Genetic Transcription; Genome engineering; Genomic approach; Goals; Heterochromatin; Histone Deacetylase; Image; Immune System Diseases; Immunologic Surveillance; Invaded; Laboratories; Lead; Leukocytes; Lighting; Link; Location; Malignant Neoplasms; Matrix Metalloproteinases; Mediating; Methods; Mitotic; Molecular; Neoplasm Metastasis; Neural Crest; Nuclear; Nuclear Hormone Receptors; Organ; Outcome; Paper; Pathogenicity; Pathology; Phase; Phenotype; Pre-Eclampsia; Process; Publishing; RNA Interference; RNA interference screen; Research; Resolution; Rheumatoid Arthritis; Structure; Testing; Therapeutic; Tissues; Treatment Efficacy; Visual; Work; Zinc Fingers; base; cdc Genes; cell behavior; cell type; design; developmental disease; developmental genetics; experimental study; functional genomics; gastrulation; genetic approach; high throughput screening; implantation; improved; in vivo; in vivo Model; innovation; insight; migration; natural Blastocyst Implantation; neoplastic cell; prevent; public health relevance; screening; stem; tool; transcription factor; transgene expression; trophoblast

Project Summary/Abstract Cell invasion through the basement membrane is a key mechanism underlying cell dispersal and organ formation during normal development, immune surveillance and is dysregulated during cancer metastasis. Yet due to difficulties of studying these dynamic behaviors in vivo, it is the least understood aspect of the metastatic cascade. My laboratory utilizes a powerful in vivo model to examine cell invasive behavior, by combining functional genomic and genetic tools with single-cell visual analyses. We examine anchor cell (AC) invasion into the vulval epithelium during C. elegans larval development. Data from our laboratory has shown a functional link between G1 phase cell cycle arrest and the acquisition of an invasive phenotype. Through high- throughput screens, we have identified that the activity of a single conserved NR2E1 class nuclear hormone receptor, the transcription factor, nhr-67 (TLX), is required to maintain the invasive AC in G1 cell cycle arrest. Loss of nhr-67 results in mitotic ACs that fail to invade. Strikingly, AC invasion can be rescued by preventing cell division through induction of G1 cell cycle arrest. Downstream of G1 arrest, chromatin modifiers, including the histone deacetylase, hda-1, are required for expression of pro-invasive genes, including matrix metalloproteinases (MMPs) and regulators of the F-actin cytoskeleton, leading to differentiation of the invasive phenotype. For this proposal we plan to elucidate how cell cycle arrest is functionally linked to invasion. In Aim 1, using molecular epistatic interaction experiments and inducible transgene expression, we will identify the upstream network of transcription factors that mediate NHR-67 activity. In Aim 2, we will generate CRISPR/Cas9-mediated conditional alleles of cell cycle control genes to determine how the AC maintains G1 arrest. In Aim 3, we will pair tissue-specific RNAi screening of chromatin modifiers with high resolution confocal and structured illumination (SIM) imaging of sub-nuclear organization, to understand the link between invasion and differentiation. The results of these aims will provide the first mechanistic view of the transcriptional and epigenetic control of cell cycle arrest and invasive behavior. Thus, our proposed work has the potential to provide a mechanistic understanding of how cells acquire and maintain an invasive phenotype, a critical aspect of many developmental and genetic disorders.

项目总结/摘要 细胞通过基底膜的侵入是细胞分散的关键机制 和器官形成过程中的正常发育，免疫监视和失调 在癌症转移过程中。然而，由于在体内研究这些动态行为的困难， 这是转移级联反应中最不为人知的方面我的实验室利用一种强大的体内 通过结合功能基因组学和遗传学工具， 单细胞视觉分析。我们检测了锚细胞（AC）侵入外阴上皮的情况 在C.线虫幼虫发育我们实验室的数据显示 G1期细胞周期停滞和获得侵袭性表型之间的关系。通过高- 通过通量筛选，我们已经确定了艾德，一个单一的保守NR 2 E1类的活性 核激素受体，转录因子，nhr-67（TLX），是维持细胞内激素水平所必需的。 侵袭性AC使G1期细胞阻滞。nhr-67的缺失导致不能侵入的有丝分裂AC。 引人注目的是，可以通过诱导G1细胞阻止细胞分裂来拯救AC入侵 循环停滞在G1期阻滞的下游，染色质修饰剂，包括组蛋白脱乙酰酶， HDA-1是表达促侵袭基因所必需的，包括基质金属蛋白酶 （MMPs）和F-肌动蛋白细胞骨架的调节因子，导致侵袭性细胞的分化。 表型对于这个建议，我们计划阐明细胞周期阻滞是如何在功能上与 入侵目的1利用分子上位性互作实验和诱导型转基因技术， 表达，我们将确定上游网络的转录因子介导的NHR-67 活动在目标2中，我们将产生CRISPR/Cas9介导的细胞周期条件等位基因， 控制基因来决定AC如何维持G1期阻滞。在目标3中，我们将组织特异性 利用高分辨率共聚焦和结构化照明筛选染色质修饰剂 (SIM)亚核组织的成像，以了解入侵和 分化这些目标的结果将提供第一个机械的观点， 转录和表观遗传控制细胞周期停滞和侵入行为。所以我们 拟议的工作有可能提供一个机制的理解细胞如何获得 并保持一种侵入性表型，这是许多发育和遗传的关键方面， 紊乱