Transcriptional regulation of morphogenetic behavior and invasive cell fate specification in C. elegans

秀丽隐杆线虫形态发生行为和侵袭细胞命运规范的转录调控

• 批准号: 9980182
• 负责人: Taylor Kinney
• 金额: $ 3.9万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-20 至 2022-07-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9980182
• 关键词: Address; BHLH Protein; Basement membrane; Behavior; Binding Sites; Biological Models; Caenorhabditis elegans; Cell Cycle; Cell Cycle Arrest; Cell Cycle Regulation; Cell Differentiation process; Cell Proliferation; Cells; Chemicals; Congenital Abnormality; Connective Tissue; Cytoskeleton; Data; Development; Disease; EVI1 gene; Epithelial; Epithelium; Event; Extracellular Matrix; Extracellular Matrix Proteins; F-Actin; Fertility; Fingers; Genes; Genetic Screening; Genetic Transcription; Genomic approach; Goals; Gonadal structure; Green Fluorescent Proteins; HELB gene; Hermaphroditism; Human; Immune System Diseases; Immune response; Immunity; Infection; Injury; Invaded; Knowledge; Leukocytes; Literature; Malignant - descriptor; Malignant Neoplasms; Matrix Metalloproteinases; Modeling; Morphogenesis; Neoplasm Metastasis; Neural Crest; Nuclear Hormone Receptors; Nuclear Localization Signal; Orthologous Gene; Outcome; Pathology; Pathway interactions; Pattern; Phenotype; Pre-Eclampsia; Pregnancy; Process; Proteins; Publishing; Puncture procedure; RARB gene; RNA Interference; Regulation; Regulator Genes; Regulatory Element; Research; Resolution; Role; Signal Transduction; Specific qualifier value; System; Testing; Therapeutic; Tissues; Transcriptional Regulation; Tubal Pregnancy; Up-Regulation; Uterus; Visual; Work; Zinc Fingers; acronyms; bZIP Domain; base; cell behavior; cell fate specification; egg; genetic approach; human DNA; improved; in vivo; notch protein; novel; programs; promoter; reverse genetics; sensor; sex; therapy development; tool; transcription factor; translational impact; trophoblast

PROJECT SUMMARY / ABSTRACT The ability of cells to traverse basement membranes (BMs) is a key part of fertility, development, immunity, and disease. BM invasion is facilitated through expression of extracellular matrix proteins, upregulation of matrix metalloproteinases, polarization of the F-actin cytoskeleton, and cell cycle arrest. Precise coordination of these pro-invasive programs is largely achieved through transcriptional regulation; however, our understanding of the gene regulatory networks (GRNs) involved is limited due to the lack of model systems in which cell invasion can be visualized live. Here, I propose to fill this gap in knowledge by utilizing morphogenesis of the Caenorhabditis elegans uterine-vulval connection as a tractable and visually amenable model to examine cell invasion in vivo. During development of the hermaphroditic somatic gonad, a specialized uterine cell called the anchor cell (AC) invades through the underlying BM to connect the uterus to the vulval epithelium. The AC itself is specified in a cell fate decision event earlier in development, in which two initially equipotent cells diverge via stochastic Notch asymmetry, giving rise to the presumptive AC and a proliferative ventral uterine (VU) cell. Prior research by our lab and others has identified six transcription factors (TFs) that regulate AC invasion. These include the basic leucine zipper TF fos-1 (Fos), the basic helix-loop-helix TF hlh-2 (E/Daughterless), two nuclear hormone receptors, nhr-67 (NR2E1/Tailless/TLX) and sex-1 (RARB/NR1B2), as well as two zinc-finger TFs, egl-43 (EVI1/MEL1) and mep-1. These TFs appear to be functioning in at least three distinct GRN sub-circuits to regulate AC invasion, one of which involves NHR-67, which functions upstream of CKI-1 (p21/p27) to induce G1 cell cycle arrest. Remarkably, five of the six pro-invasive TFs function reiteratively during the AC/VU cell fate decision. These include the three TFs comprising the NHR-67/cell cycle-dependent pro-invasive pathway (EGL- 43S, MEP-1, and NHR-67), as well as HLH-2 and SEX-1, which have predicted binding sites within the nhr-67 promoter. Thus, based on the literature and my preliminary studies, my central hypothesis is that the AC invasive program is dependent on the function of multiple GRN sub-circuits, one of which modulates cell cycle arrest and is reiteratively used in AC fate specification. In Aim 1 of this project, I will dissect the cis- and trans-regulation of AC invasion, focusing on the cell cycle-dependent GRN sub-circuit involving the pro-invasive TF nhr-67/TLX. In Aim 2, I will examine the roles of pro-invasive TFs that reiteratively function in AC specification and investigate if cell cycle control is the common denominator underlying these two processes. Cutting-edge functional tools, including an endogenous protein depletion system and a novel cell cycle state sensor, paired with the ability to perform high-resolution subcellular visual analyses, will allow for thorough and rigorous testing of this hypothesis.

项目摘要 /摘要 细胞遍布基底膜（BMS）的能力是生育，发育，免疫力和 疾病。通过表达细胞外基质蛋白，基质的上调来促进BM侵袭 金属蛋白酶，F-肌动蛋白细胞骨架的极化和细胞周期停滞。这些的精确协调 亲侵入性计划在很大程度上是通过转录调节实现的；但是，我们对 由于缺乏模型系统，涉及的基因调节网络（GRN）受到限制 可视化现场。在这里，我建议通过利用Caenorhabditis的形态发生来填补这一知识的空白。 秀丽隐杆线虫的子宫浮雕连接是一种可进行的且视觉上可正约的模型，可检查体内细胞侵袭。 在雌雄同体体细胞性腺的发育过程中，一个专门的子宫细胞称为锚碱细胞（AC） 通过基础BM入侵，将子宫连接到外阴上皮。交流本身在 细胞命运决策事件早期开发中，其中两个最初的等值细胞通过随机缺口发散 不对称，引起推定的AC和增殖性腹侧子宫（VU）细胞。我们的事先研究 实验室和其他人确定了调节AC侵袭的六个转录因子（TFS）。这些包括基本 亮氨酸拉链TF FOS-1（FOS），基本的螺旋环 - 螺旋TF HLH-2（E/Daumponless），两个核激素 受体，NHR-67（NR2E1/TAILLESS/TLX）和SEX-1（RARB/NR1B2），以及两个锌指TFS，EGL-43 （EVI1/MEL1）和MEP-1。这些TF似乎在至少三个不同的GRN子电路中起作用 调节AC入侵，其中之一涉及NHR-67，它在CKI-1（P21/P27）上游起作用以诱导G1 细胞周期停滞。值得注意的是，在AC/VU细胞命运期间，六个亲侵入性TFS中有五个重复性 决定。其中包括包含NHR-67/细胞周期依赖性侵入性途径的三个TF（EGL- 43S，MEP-1和NHR-67）以及HLH-2和SEX-1，它们预测了NHR-67内的绑定位点 发起人。因此，基于文献和我的初步研究，我的中心假设是AC侵入性 程序取决于多个GRN亚电路的功能，其中之一调节细胞周期停滞和 重申用于AC命运规范。在该项目的目标1中，我将剖析顺式和反式调节 AC侵袭，重点是涉及侵入性TF NHR-67/TLX的细胞周期依赖性GRN亚电路。在 AIM 2，我将研究重申在AC规范中重复起作用并研究的亲侵入性TF的作用 如果细胞周期控制是这两个过程的基础分母。尖端功能工具， 包括内源性蛋白质耗竭系统和新型细胞周期态传感器，并与能够 进行高分辨率亚细胞视觉分析，将允许对该假设进行彻底和严格的测试。