Investigating the molecular and mechanical regulation of pulsed actomyosin contra

研究脉冲肌动球蛋白拮抗剂的分子和机械调节

基本信息

批准号：
7770569
负责人：
Adam Christopher Martin
金额：
$ 8.58万
依托单位：
PRINCETON 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：
2010-01-15 至 2010-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7770569
关键词：
Ablation Abnormal Cell Actins Actomyosin Apical Architecture Automobile Driving Biochemical Biochemistry Biology Cell Shape Cells Cellular biology Complex Cytoskeletal Modeling Cytoskeleton Data Development Developmental Cell Biology Drosophila genus Education Environment Epithelial Epithelial Cells Epithelium Foundations Future Gene Expression Generations Genes Genetic Goals Homologous Gene Human Image Analysis Individual Lasers Learning Life Malignant Neoplasms Measures Mechanics Mentors Mesoderm Cell Microfilaments Modeling Molecular Morphogenesis Motor Activity Myosin Type II Neoplasm Metastasis Organ Pharmaceutical Preparations Phase Photobleaching Physics Physiologic pulse Play Postdoctoral Fellow Process Property Proteins RNA Interference Regulation Research Resources Retinal Cone Role Running Shapes Signal Transduction Snails Stretching System Technical Expertise Testing Time Tissues Training Translating Universities Variant Work cancer cell cell behavior cell growth cellular imaging constriction driving force experience gastrulation graduate student improved insight interdisciplinary approach mathematical model multidisciplinary neoplastic cell new therapeutic target photoactivation prevent prospective protein function public health relevance research study skills transcription factor tumor progression

项目摘要

DESCRIPTION (provided by applicant): Morphogenesis is the process whereby simple tissues, such as epithelial sheets, are sculpted into complex organs. Morphogenesis is driven by forces generated by individual cells, which result in changes in cell shape and tissue mechanics. During development, these changes are tightly regulated in space and time by both genetic and mechanical signals. During cancer, these signals are often improperly activated, resulting in abnormal cell behavior that leads to tumor cell growth and metastasis. Therefore, understanding how cells and tissues generate forces is essential to understand development and cancer. Because morphogenesis depends on the complex interplay of molecular and mechanical signals, identifying the mechanisms that drive morphogenesis requires a multidisciplinary approach that includes biochemistry, genetics, cell and developmental biology, physics, and mathematical modeling. As a graduate student in David Drubin's lab at UC Berkeley, I was trained in cell biology, biochemistry, and genetics. Specifically, I gained much experience working with the actin cytoskeleton, which generates mechanical forces in cells. As a postdoctoral fellow in Eric Wieschaus' lab at Princeton University, I have learned Drosophila biology and have begun to develop quantitative and computational skills to analyze the dynamics of multicellular systems. Specifically, I have analyzed apical constriction, a common cell shape change that facilitates epithelial bending and tissue invagination. These complementary research experiences provide me with a unique perspective and a range of technical expertise that I will use in my independent lab to study how the actin cytoskeleton generates forces during development. In the Wieschaus lab, I discovered that apical constriction is driven by pulsed actomyosin contractions, which incrementally constrict the cell. Pulsed contractions are regulated by the transcription factors Twist and Snail, whose human homologues play important roles in cancer cell metastasis. In the current research plan, I propose experiments that will elucidate the mechanisms that regulate pulsed contraction. This will be achieved by integrating live-cell imaging, quantitative image analysis, genetics, biochemistry, and mathematical modeling. One goal will be to identify the molecular mechanisms that control pulsed contractions downstream of the transcription factors Twist and Snail. A second goal will be to determine how mechanical forces transmitted through the tissue regulate cell shape change and cytoskeletal organization during morphogenesis. To accomplish the goals of my proposal, I need additional training in quantitative image analysis, mathematical modeling, and physics. This will allow me to more effectively analyze the dynamics of the actin cytoskeleton and the physical interactions between cells in multicellular systems, which will be essential foundations for my future independent lab. The Wieschaus lab is the ideal environment to obtain this training because we are part of the Center for Quantitative Biology at Princeton University. Eric Wieschaus is an excellent mentor who strongly believes in quantifying experimental data and developing quantitative models to explain this data. I also collaborate with a theoretical physicist at Princeton, Matthias Kaschube, who is an expert on quantitative image analysis. Furthermore, Princeton offers a variety of seminars, classes, and resources that are at my disposal to further my education in quantitative biology. The additional training I obtain at Princeton will greatly improve my skills in quantitative analysis and modeling, and will increase the quality and impact of my future research. Overall, this experience will help me achieve my goal of running a multidisciplinary lab that performs cutting edge research on morphogenesis. Public Health Relevance: During development and cancer progression, gene expression induces mechanical changes in cells that result in changes in cell shape and tissue architecture. We will investigate the function of two genes that promote cell shape changes during the development of the fruit fly, and whose human homologues are involved in cancer cell metastasis. We will investigate how these genes generate forces in cells and tissues and whether the mechanical forces in a tissue regulate individual cell behavior.

描述（由申请人提供）：形态发生是简单组织（例如上皮片）被塑造成复杂器官的过程。形态发生是由单个细胞产生的力驱动的，这会导致细胞形状和组织力学的变化。在发育过程中，这些变化在空间和时间上受到遗传和机械信号的严格调控。在癌症过程中，这些信号常常被不当激活，导致细胞行为异常，从而导致肿瘤细胞生长和转移。因此，了解细胞和组织如何产生力对于了解发育和癌症至关重要。由于形态发生取决于分子和机械信号的复杂相互作用，因此识别驱动形态发生的机制需要采用多学科方法，包括生物化学、遗传学、细胞和发育生物学、物理学和数学建模。作为加州大学伯克利分校 David Drubin 实验室的研究生，我接受了细胞生物学、生物化学和遗传学方面的培训。具体来说，我在肌动蛋白细胞骨架方面获得了很多经验，它在细胞中产生机械力。作为普林斯顿大学 Eric Wieschaus 实验室的博士后研究员，我学习了果蝇生物学，并开始发展定量和计算技能来分析多细胞系统的动力学。具体来说，我分析了顶端收缩，这是一种常见的细胞形状变化，有助于上皮弯曲和组织内陷。这些互补的研究经验为我提供了独特的视角和一系列技术专业知识，我将在我的独立实验室中使用它们来研究肌动蛋白细胞骨架在发育过程中如何产生力。在 Wieschaus 实验室，我发现顶端收缩是由脉冲肌动球蛋白收缩驱动的，它会逐渐收缩细胞。脉冲式收缩由转录因子 Twist 和 Snail 调节，它们的人类同源物在癌细胞转移中发挥重要作用。在当前的研究计划中，我提出了一些实验来阐明调节脉冲收缩的机制。这将通过整合活细胞成像、定量图像分析、遗传学、生物化学和数学建模来实现。目标之一是确定控制转录因子 Twist 和 Snail 下游脉冲收缩的分子机制。第二个目标是确定通过组织传递的机械力如何在形态发生过程中调节细胞形状变化和细胞骨架组织。为了实现我的提案的目标，我需要在定量图像分析、数学建模和物理学方面进行额外的培训。这将使我能够更有效地分析肌动蛋白细胞骨架的动力学以及多细胞系统中细胞之间的物理相互作用，这将是我未来独立实验室的重要基础。 Wieschaus 实验室是获得这种培训的理想环境，因为我们是普林斯顿大学定量生物学中心的一部分。 Eric Wieschaus 是一位优秀的导师，他坚信量化实验数据并开发定量模型来解释这些数据。我还与普林斯顿大学的理论物理学家 Matthias Kaschube 合作，他是定量图像分析方面的专家。此外，普林斯顿大学还提供各种研讨会、课程和资源，可供我使用以进一步加深我在定量生物学方面的教育。我在普林斯顿大学获得的额外培训将极大地提高我在定量分析和建模方面的技能，并将提高我未来研究的质量和影响力。总的来说，这段经历将帮助我实现运营一个多学科实验室的目标，该实验室对形态发生进行前沿研究。公共健康相关性：在发育和癌症进展过程中，基因表达会诱导细胞发生机械变化，从而导致细胞形状和组织结构发生变化。我们将研究果蝇发育过程中促进细胞形状变化的两个基因的功能，它们的人类同源物参与癌细胞转移。我们将研究这些基因如何在细胞和组织中产生力，以及组织中的机械力是否调节个体细胞的行为。