A Molecular Toolkit for Controlling and Probing Cell Junction-Actin Interactions

用于控制和探测细胞连接-肌动蛋白相互作用的分子工具包

• 批准号: 10276194
• 负责人: Brian Belardi
• 金额: $ 38.06万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10276194
• 关键词: Actins; Adhesions; Award; Binding; Biological; Biology; Cadherins; Carcinoma; Cell Shape; Cells; Characteristics; Communities; Cytoskeleton; Development; Disease; Disease Progression; Elements; Endocytosis; Engineering; Epithelial; Exhibits; Extracellular Matrix; Filament; Focal Adhesions; Glues; Human; Individual; Integrins; Intercellular Junctions; Link; Maintenance; Malignant Neoplasms; Mechanics; Membrane; Membrane Biology; Membrane Proteins; Methods; Microfilaments; Mitosis; Molecular; Morphogenesis; Natural Products; Organism; Permeability; Play; Polymers; Process; Property; Proteins; Research; Research Personnel; Role; Side; Signal Transduction; Specificity; Structure; System; Tight Junctions; Tissues; Translational Research; Work; base; cell motility; inhibitor/antagonist; innovation; interest; mechanical properties; monolayer; programs; reconstitution; repaired; side effect; small molecule; therapeutic target; tool; vector

PROJECT SUMMARY: A MOLECULAR TOOLKIT FOR CONTROLLING AND PROBING CELL JUNCTION- ACTIN INTERACTIONS Higher metazoans exhibit robust, yet dynamic connections between neighboring cells, leading to the exquisite morphogenesis, vectorial transport, and resilient mechanical properties that define tissue. Spatially separated junctions line individual epithelial membranes and are tasked with linking cells to one another and to the underlying extracellular matrix. These junctions are composed of well-characterized membrane proteins, each with unique functions: claudins create paracellular barriers; cadherins bind cells together; and integrins attach cells to matrix. Despite unique classes of membrane proteins, different junctions all possess a common element, the cytoskeleton, which resides on the cytosolic side of the contact. One cytoskeletal polymer in particular – actin – appears indispensable for junction activity. While decades of elegant work have transformed our understanding of the structure and binding characteristics of junctional membrane proteins, the question of how actin is involved in cell junction formation, junction maintenance and repair, and junctional signaling remains unresolved. Actin filaments are ubiquitous throughout the cell as they contribute to cell shape, endocytosis, mitosis, motility, and other critical phenomena. However, this wide distribution presents a fundamental problem when studying actin – namely how to pinpoint the exact role actin filaments play in the process-of-interest. While actin- targeted natural products and small molecules are widely used to disrupt filaments globally, they lack the specificity needed to uncover the role of actin filaments locally at cell junctions. My research group is developing a suite of molecular tools to both control and dissect actin interactions at cell junctions. In this way, we provide researchers with new methods to turn-on and -off actin association and to probe actin’s role in adhesion and cell-cell mechanics. These tools come in various molecular forms: i) protein-based switches, ii) small-molecule molecular glues and inhibitors, and iii) synthetic cells, which can be applied to wide-ranging systems, such as reconstituted membranes, cells, monolayers, tissues, and organisms, to illuminate and manipulate actin- dependent processes. In my lab, we will harness these molecular tools to focus on three specific research directions in epithelial biology, although we anticipate that the toolkit will benefit the greater biological community, including biochemists, cell biologists and developmental biologists. First, we will focus on applying our tools to dissect actin’s role during tight junction maturation and, ultimately, to modulate barrier function. Second, we will investigate, in mechanistic detail, how actin potentiates integrin activation during focal adhesion formation. Finally, we will assemble cells using actin switches to generate “synthetic tissues” with programmable and toggleable properties, such as dynamic tissue permeability and adhesion. Broadly, this research program relies on our diverse expertise in molecular engineering, basic membrane biology, and translational science to create a virtuous cycle of innovation and discovery over the course of the MIRA award.

项目摘要：用于控制和探测细胞连接的分子工具包 - 肌动蛋白相互作用 较高的后生动物暴露了稳健但动态的连接，导致了独家 定义组织的形态发生，媒介传输和弹性的机械性能。空间分开 连接线单个上皮膜，其任务是将细胞相互联系，并与 潜在的细胞外基质。这些连接由良好的膜蛋白组成，每个膜蛋白 具有独特的功能：Claudins会产生细胞细胞壁屏障；钙粘蛋白结合细胞在一起；和整合素 细胞到矩阵。尽管膜蛋白具有独特的类别，但不同的连接都具有共同的元素， 细胞骨架位于接触的胞质侧。特别是一种细胞骨架聚合物 - 肌动蛋白 - 对于连接活动而言似乎是必不可少的。而数十年的优雅作品改变了我们的理解 连接膜蛋白的结构和结合特征，肌动蛋白如何涉及的问题 在细胞连接形成中，连接维护和修复以及连接信号传导仍未解决。 肌动蛋白丝在整个细胞中无处不在，因为它们有助于细胞形状，内吞作用，有丝分裂， 运动和其他关键现象。但是，这种广泛的分布在 研究肌动蛋白 - 即如何查明肌动蛋白丝在利益过程中起着确切的作用。而肌动蛋白 - 靶向天然产物和小分子被广泛用于全球破坏细丝，它们缺乏 揭示肌动蛋白丝在细胞连接处的作用所需的特异性。我的研究小组正在发展 一组分子工具，用于控制和剖析细胞连接处的肌动蛋白相互作用。这样，我们提供 具有新方法的研究人员，可以进行转动和 - 抗肌动蛋白协会，并探讨肌动蛋白在粘合剂中的作用 细胞电池力学。这些工具有各种分子形式：i）基于蛋白质的开关，ii）小分子 分子胶和抑制剂，以及iii）合成细胞，可以应用于大型系统，例如 重建的机制，细胞，单层，组织和生物，以照亮和操纵肌动蛋白 依赖过程。 在我的实验室中，我们将利用这些分子工具专注于上皮的三个特定研究方向 生物学，尽管我们预计该工具包将受益于更大的生物学界，包括 生化主义者，细胞生物学家和发育生物学家。首先，我们将专注于应用我们的工具进行剖析 肌动蛋白在紧密结合成熟过程中的作用，并最终调节屏障功能。第二，我们会的 从机械的细节中，肌动蛋白如何增强整联蛋白在局灶性形成过程中如何增强整联蛋白的活化。 最后，我们将使用肌动蛋白开关组装细胞，以通过可编程和可编程生成“合成组织” 可切换的特性，例如动态组织渗透性和粘合剂。从广义上讲，该研究计划依赖 关于分子工程，基本膜生物学和转化科学领域的潜水专业知识 在MIRA奖的过程中，创新和发现的虚拟循环。