Investigating mechanisms regulating cytoskeletal dynamics and alignment during epithelial tissue folding

研究上皮组织折叠过程中细胞骨架动力学和排列的调节机制

• 批准号: 10396453
• 负责人: Mary Ann Collins
• 金额: $ 6.76万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10396453
• 关键词: Actomyosin; Adherens Junction; Adult; Affect; Apical; Apoptosis; Automobile Driving; Behavior; Cancer Biology; Cell Shape; Cells; Cellular biology; Chemicals; Collection; Congenital Abnormality; Connective Tissue; Cytoskeletal Modeling; Cytoskeleton; Defect; Deformity; Development; Developmental Biology; Disease; Disease Progression; Disputes; Drosophila genus; Drosophila melanogaster; Embryo; Embryonic Development; Epithelial; Event; F-Actin; Failure; Fogs; Gene Expression; Genetic; Goals; Health; Human; Image; Image Analysis; Individual; Intercellular Junctions; Knowledge; Label; Lead; Light; Link; Malignant Neoplasms; Measures; Mechanics; Mesenchymal; Mesoderm; Methods; Microfilaments; Microscopy; Mission; Molecular; Molecular Motors; Morphogenesis; Motor; Movement; Myosin ATPase; Myosin S-2; Myosin Type II; Nature; Neoplasm Metastasis; Neural Tube Defects; Neural tube; Organ; Outcome; Physiologic pulse; Process; Production; Research; Resolution; Role; Seminal; Shapes; Signal Pathway; Signal Transduction; Structure; Surface; TWIST1 gene; Testing; Tissues; United States National Institutes of Health; Work; alpha catenin; base; comparative; constriction; crosslink; developmental disease; driving force; epithelial to mesenchymal transition; gastrulation; imaging modality; innovation; insight; intercellular connection; mechanical force; mechanical signal; mutant; myosin phosphatase; non-muscle myosin; novel; physical property; predictive modeling; prevent; quantitative imaging; single molecule; training opportunity; transmission process; tumor progression

Project Summary: Large-scale tissue movements are critical during development to transform an amorphous collection of cells into organs with specific structure and function. Abnormal activation of force-generating signals that regulate epithelial morphogenesis can result in developmental defects, such as neural tube deformities, as well as aberrant epithelial-mesenchymal transition and cancer metastasis. Yet we do not fully understand how mechanical forces generated at the molecular level regulate epithelial remodeling. At the cellular level, most forces are generated by the actomyosin network; the molecular motor non-muscle myosin II (myosin) crosslinks actin filaments (F-actin), thereby generating contractile forces which are propagated throughout the tissue via intercellular connections. One outcome of actomyosin contractility is apical contraction, in which the apex of the cell narrows as a result of repeated bursts of myosin pulses that condense the F-actin cortex in a ratchet-like manner. When myosin pulsing and ratcheting is disrupted, cells fail to apically constrict and the tissue fails to fold. However, the mechanisms driving pulsatile contractions and ratcheting behavior remains poorly understood, highlighting a critical gap in our understanding of how upstream signaling events are intricately linked to downstream changes in cytoskeletal organization and behavior. The long-term goal of this project is to determine how mechanical forces generated at the molecular level collectively drive tissue-wide morphogenetic changes. The overall objective of this proposal is to identify mechanisms that regulate myosin dynamics and alignment by determining the mechanistic link between Twist expression and myosin turnover. The rationale for this proposed work is to gain insight not only into the nature of these mechanisms, but also the general principles governing contractility and ratchet-like apical constriction during large-scale tissue movements. Our central hypothesis is that Twist, and its downstream effectors, as well as tissue-wide forces, via intercellular connections, cooperatively regulate myosin dynamics to drive apical ratcheting and tissue remodeling events during embryonic development in Drosophila. This hypothesis will be tested by pursuing two specific aims: we will (1) determine the mechanism through which Twist promotes cell apex stabilization, and (2) determine how myosin dynamics are affected by intercellular connectivity. Our approach is innovative because it is one of the first to directly examine myosin dynamics using an integrative strategy that combines classic Drosophila genetics with advanced microscopy methods, including photo-conversion and super- resolution imaging. The proposed research is significant because it will advance our understanding of the connection between gene expression, signaling pathways, and force production during epithelial morphogenesis, and will provide new perspective to ongoing research efforts investigating developmental diseases and cancer biology.

项目总结： 大规模的组织运动在发育过程中是至关重要的，以改变无定形的 细胞分化成具有特定结构和功能的器官。力产生信号的异常激活 调节上皮形态发生也会导致发育缺陷，如神经管畸形。 异常上皮-间充质转化和癌症转移。然而，我们并不完全了解 在分子水平上产生的机械力调节上皮重塑。在细胞层面上，大多数 肌球蛋白网络产生力量；分子马达非肌肉肌球蛋白II(Myosin) 交联化肌动蛋白细丝(F-肌动蛋白)，从而产生收缩力量，并在整个 通过细胞间连接的组织。肌动球蛋白收缩的一个结果是心尖收缩，在这种情况下 细胞的顶端由于肌球蛋白脉冲的反复爆发而变窄，肌球蛋白脉冲将F-肌动蛋白皮质浓缩在 棘轮式的方式。当肌球蛋白的脉冲和棘轮被破坏时，细胞不能顶端收缩， 组织不能折叠。然而，驱动搏动性收缩和棘轮行为的机制仍然存在 知之甚少，突显出我们对上游信令事件的理解存在严重差距 错综复杂地与细胞骨架组织和行为的下游变化有关。这样做的长期目标是 该项目是为了确定在分子水平上产生的机械力如何共同驱动整个组织 形态发生变化。这项提案的总体目标是确定调节肌球蛋白的机制 通过确定扭转表达和肌球蛋白周转之间的机制联系来实现动力学和排列。 这项拟议工作的基本原理不仅是为了深入了解这些机制的性质，而且是为了 大范围组织收缩和棘齿状根尖收缩的一般规律 动静。我们的中心假设是Twist和它的下游效应器，以及组织范围的力， 通过细胞间连接，协同调节肌球蛋白动力学以驱动尖端棘轮和组织 果蝇胚胎发育中的重塑事件。这一假设将通过追求两个 具体目标：我们将(1)确定Twist促进细胞顶端稳定的机制，以及 (2)确定细胞间连通性对肌球蛋白动力学的影响。我们的方法是创新的 因为它是第一个使用综合策略直接检查肌球蛋白动力学的公司之一， 经典的果蝇遗传学和先进的显微镜方法，包括光转化和超微结构 分辨率成像。这项拟议的研究具有重要意义，因为它将促进我们对 上皮细胞中基因表达、信号通路和力产生之间的联系 形态发生，并将为正在进行的研究开发提供新的视角 疾病和癌症生物学。