NanoPlan - Deciphering the molecular construction rules of integrin-based cell adhesions.

NanoPlan - 破译基于整合素的细胞粘附的分子构建规则。

• 批准号: 316712267
• 负责人: Professor Dr. Carsten Grashoff
• 金额: --
• 依托单位: Institut für Integrative Zellbiologie und Physiologie (IIZP)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/316712267?language=en
• 关键词: NanoPlan; Deciphering; molecular; construction; rules

Cell adhesion to the extracellular matrix (ECM) is fundamental to many developmental and pathophysiological processes. It is mediated by integrin receptors that bind to a wide range of extracellular ligands and intracellularly connect to the actin cytoskeleton in macromolecular structures called focal adhesions (FAs). FAs are highly complex assemblies containing hundreds of different proteins processing chemical as well as mechanical signals during cell adhesion; however, the molecular processes that govern FA organization and mechano-chemical signaling remain incompletely understood. Furthermore, mammals express 24 distinct integrin receptors with specific ligand binding and signaling properties but the underlying mechanisms mediating integrin receptor specificity are still unclear. The major obstacle for a better understanding of integrin function has been the lack of suitable techniques that allow a systematic investigation of FAs with sufficient temporal and spatial resolution. We, the members of this French-German Consortium, have therefore developed techniques that will allow such an analysis. We have established unique cell systems to study distinct integrin receptors and their intracellular regulators at physiological expression levels and we have developed super-resolution microscopy methods to study the dynamic nanoscale organization of individual FAs proteins. In addition, we have developed mass-spectrometry approaches to study the molecular composition of FAs and engineered biosensors to quantify integrin force transduction with piconewton sensitivity in cells. We propose to combine our tools to investigate how integrins and their associated molecules regulate the nanoscale organization of FAs to modulate chemical and mechanical signaling during cell adhesion in an integrin subtype specific fashion.In the first part of the project, single-protein tracking and super-resolution microscopy will be used to determine the dynamics of distinct integrin subunits in genetically modified cells. We will combine super-resolution microscopy with nano-patterning to control the lateral spacing of integrin subtype specific ligands and apply molecular tension sensors to correlate FA nanoscale organization with integrin force transduction. In the second part, we will analyze the role of integrin regulators on FA nanoscale organization, molecular dynamics and force transduction. Finally, we will evaluate how externally applied forces are processed by distinct integrin receptors and their associated molecules. These experiments will utilize custom-built cell stretchers that allow super-resolution microscopy as well as molecular force measurements in living cells. Together, the proposed experiments should provide unprecedented insights into the molecular construction rules of FAs. The expected results should be valuable to the cell adhesion community and be of general interest to the cell biological and biophysical community.

细胞粘附到细胞外基质（ECM）是许多发育和病理生理过程的基础。它是由整合素受体介导的，整合素受体与广泛的细胞外配体结合，并在细胞内与称为粘着斑（FA）的大分子结构中的肌动蛋白细胞骨架连接。FA是高度复杂的组件，包含数百种不同的蛋白质处理化学以及细胞粘附过程中的机械信号;然而，管理FA组织和机械化学信号的分子过程仍然不完全清楚。此外，哺乳动物表达24种具有特异性配体结合和信号传导特性的不同整合素受体，但介导整合素受体特异性的潜在机制仍不清楚。更好地了解整合素功能的主要障碍是缺乏合适的技术，使系统的调查FA具有足够的时间和空间分辨率。因此，我们法德联合会的成员开发了能够进行这种分析的技术。我们已经建立了独特的细胞系统来研究不同的整合素受体及其细胞内调节因子在生理表达水平，我们已经开发出超分辨率显微镜方法来研究动态纳米级组织的个别脂肪酸蛋白。此外，我们还开发了质谱方法来研究脂肪酸和工程生物传感器的分子组成，以量化整合素力转导与细胞中的皮牛顿灵敏度。我们建议联合收割机我们的工具来研究整合素及其相关分子如何调节脂肪酸的纳米级组织，以调节细胞粘附过程中的化学和机械信号，在整合素亚型特异性fashion.In第一部分的项目，单蛋白跟踪和超分辨率显微镜将被用来确定不同的整合素亚基在转基因细胞的动力学。我们将结合联合收割机超分辨率显微镜与纳米图案化来控制整合素亚型特异性配体的横向间距，并应用分子张力传感器将FA纳米级组织与整合素力转导相关联。在第二部分中，我们将分析整合素调节剂在FA纳米级组织、分子动力学和力传递中的作用。最后，我们将评估外部施加的力是如何处理不同的整合素受体及其相关分子。这些实验将利用定制的细胞拉伸器，允许超分辨率显微镜以及活细胞中的分子力测量。总之，拟议的实验应该提供前所未有的深入了解FA的分子结构规则。预期的结果对细胞粘附界应该有价值，并引起细胞生物学和生物物理界的普遍兴趣。