Investigating the mechanism of self-organized cortical patterning in an artificial cortex

研究人工皮质中自组织皮质模式的机制

• 批准号: 10656543
• 负责人: Jennifer Elaine Landino
• 金额: $ 0.77万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10656543
• 关键词: Actomyosin; Aneuploidy; Architecture; Award; Biochemical; Biochemical Feedback; Cell Cycle; Cell Shape; Cell division; Cell membrane; Cell model; Cell physiology; Cell surface; Cell-Free System; Cells; Chromosomes; Complex; Cytokinesis; Cytoskeletal Modeling; Cytoskeleton; Data; Development; Disease; Electron Microscopy; Embryo; Ensure; Environment; Equity; F-Actin; Faculty; Failure; Feedback; Goals; Guanosine Triphosphate Phosphohydrolases; Individual; Knowledge; Laboratory Research; Lipid Bilayers; Lipids; Liquid substance; Malignant Neoplasms; Mediating; Membrane; Membrane Fluidity; Michigan; Microtubules; Modeling; Molecular; Monomeric GTP-Binding Proteins; Pattern; Pattern Formation; Polymers; Positioning Attribute; Preparation; Process; Proteins; Publishing; Regulation; Regulation of Cell Shape; Research; Role; Shapes; Signal Transduction; Signaling Molecule; Signaling Protein; Structural Protein; Structure; Surface; System; Testing; Training; Training Support; Universities; Work; Workplace; Xenopus; cancer cell; career; cell cortex; cell motility; egg; flexibility; fluidity; in vivo; migration; novel; polymerization; programs; reconstitution; rho; self organization; spatiotemporal; success; two-dimensional

The cell cortex underlies essential cellular functions, including cell shape changes that facilitate cell division. Comprised of a meshwork of filamentous actin (F-actin) and the plasma membrane, the cortex is remodeled during cytokinesis, physically dividing the cell in two. Recent work has shown that prior to large-scale remodeling, the cortex is also dynamically patterned with coherent subcellular waves of the small GTPase RhoA and F-actin, a phenomenon termed “cortical excitability”. In developing embryos, these waves appear over the entire surface of the cell and then feed into the cytokinetic furrow as cell division progresses and have been proposed to support the rapid and flexible establishment of the division plane. Investigating the mechanisms that support and regulate cortical patterning is currently limited by a lack of technical approaches that can bridge our understanding of biochemical feedback signaling and cortical pattern formation, including the molecular regulation of signaling molecules, membrane dynamics, and cytoskeletal remodeling. A breakthrough in this gap in knowledge has been the development by the applicant of an “artificial cortex”, made from supported lipid bilayers (SLBs) and Xenopus egg extract, which successfully reconstitutes active Rho and F-actin dynamics in a cell-free system. Like in vivo cortical excitability, patterning in the artificial cortex depends on Rho activity and F-actin polymerization. This novel, synthetic approach to investigating cortical patterning is an ideal system for systematically examining the role of individual factors (such as upstream GTPase regulators, membrane composition and fluidity, cell cycle state) in regulating cortical dynamics. Using the artificial cortex as a model for cortical patterning, this proposal for a MOSAIC K99/R00 Award seeks to understand how cortical pattern formation is regulated and how patterning remodels the cell cortex to perform essential functions like cytokinesis. Dr. Landino will investigate the factors that drive cortical wave formation (Aim 1), cytoskeletal remodeling at the cortex (Aim 2), and the role of cortical patterning in supporting successful cell division (Aim 3). The results of this work will expand our knowledge of the molecular regulation of the cortex underlying the emergence of cortical excitability, and the role of dynamic patterning in cell division. Dr. Landino's long-term career goal is to establish an independent research group investigating the mechanisms that regulate cortical patterning and cell division. The proposed training will provide Dr. Landino with additional scientific expertise, including technical training in electron microscopy and preparation of cycling extract, and further establish the artificial cortex as a useful platform for understanding the biochemical and structural regulation of the cell cortex. This award will further Dr. Landino's professional development including formal training in research laboratory management, leading a diverse, equitable, and inclusive workplace, and a tailored plan to support Dr. Landino's application to faculty positions. The exemplary scientific and professional environment at the University of Michigan is ideally suited to support the training outlined in this proposal and ensure Dr. Landino's success in launching an independent research program.

细胞皮层是重要细胞功能的基础，包括促进细胞分裂的细胞形状变化，皮层由丝状肌动蛋白（F-肌动蛋白）和质膜组成，在胞质分裂过程中进行重塑，将细胞物理分成两部分。研究表明，在大规模重塑之前，皮质也会动态地形成小 GTP 酶 RhoA 和 F-肌动蛋白的连贯亚细胞波，这种现象在发育中的胚胎中被称为“皮质兴奋性”。这些波出现在细胞的整个表面，然后随着细胞分裂的进展进入细胞分裂沟，并被提议支持分裂平面的快速和灵活的建立。目前，研究支持和调节皮质模式的机制受到限制。缺乏能够弥合我们对生化反馈信号传导和皮质模式形成的理解的技术方法，包括信号分子的分子调节、膜动力学和细胞骨架重塑。申请人开发了一种技术，解决了这一知识空白。 “人造的“皮质”，由支持的脂质双层 (SLB) 和爪蟾卵提取物制成，成功地在无细胞系统中重建活性 Rho 和 F-肌动蛋白动力学，与体内皮质兴奋性一样，人工皮质中的模式取决于 Rho 活性和 F-肌动蛋白。这种研究皮质模式的新颖合成方法是系统研究单个因素（例如上游 GTP 调节因子、膜组成和流动性、细胞周期状态）在调节中的作用的理想系统。 Landino 博士使用人造皮质作为皮质图案化模型，旨在了解皮质图案形成是如何调节的以及图案化如何重塑细胞皮质以执行胞质分裂等基本功能。研究驱动皮质波形成的因素（目标 1）、皮质细胞骨架重塑（目标 2）以及皮质模式在支持成功细胞分裂中的作用（目标 3）。这项工作的结果将扩大我们对皮层兴奋性出现背后的皮层分子调节的认识，以及动态模式在细胞分裂中的作用。兰迪诺博士的长期职业目标是建立一个独立的研究小组来研究其机制。拟议的培训将为兰迪诺博士提供额外的科学专业知识，包括电子显微镜和循环提取物制备方面的技术培训，并进一步将人工皮层建立为了解生化和结构调节的有用平台。细胞的该奖项将促进兰迪诺博士的专业发展，包括研究实验室管理方面的正式培训，领导一个多元化、公平和包容的工作场所，以及支持兰迪诺博士申请教职的定制计划。密歇根大学非常适合支持本提案中概述的培训，并确保兰迪诺博士成功启动独立研究项目。