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Coupling Force, Tension and Cell Plasma Membrane Plasticity at the Nanoscale Functional Roles of Caveolin Nanodomains

Caveolin 纳米域的纳米级功能作用中的耦合力、张力和细胞质膜可塑性

基本信息

批准号：
1806381
负责人：
Fabien Pinaud
金额：
$ 42万
依托单位：
University of Southern California
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-15 至 2022-08-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1806381&HistoricalAwards=false
关键词：
Coupling Force Tension Cell Plasma

项目摘要

Mechanical forces exerted at the cell plasma membrane (PM) direct many important cellular and tissue processes, including cell adhesion, migration and invasiveness. In these processes, the membrane curving protein caveolin 1 (cav1) and the different types of nano-scale domains it forms at the PM are emerging as critical mechano-transducing hubs and tension buffering structures of mammalian cells. Yet, the structural plasticity of cav1 nano-scale domains and their functional coupling to the local mechanical and tension states of the PM are still enigmatic. Consequently, some of the fundamental mechanisms by which the cell membrane adapts to varying forces at the nano-scale remain undefined. This project aims at understanding how cav1 nano-scale domains modulate the PM structure in response to mechanical cues and at defining the core physical principles that govern PM plasticity, maintenance of membrane tension and proper cell response to forces. This project will be implemented through a multidisciplinary approach that integrates super-resolution (SR) microscopy imaging, optical force sensing, engineering of bio-materials for cell mechano-biology, quantitative biophysics and modeling. It will (i) define, quantitatively, the nano-scale organization of cav1 nano-scale domains at the cell PM, (ii) establish their plasticity in response to specific PM forces, (iii) determine how they locally regulate PM tension and (iv) provide physical models of their functions as key PM tension modulators. By its original and multidisciplinary nature, the project engages its participants in highly interdisciplinary research. It will provide them with skills that match the current convergence between cell biology, physics and engineering research. The proposed activities also provide a platform integrating modern technology and cross-pollination science for traditionally underrepresented students at the undergraduate and high school levels, through (i) hands-on experimentation with imaging probes, cell culture and microscopy imaging, (ii) outreach projects and (iii) educational activities. Economically disadvantaged undergraduate and high school students will be recruited via established programs at USC and through outreach to high schools in the Greater Los Angeles Area. In particular, an 8-weeks Summer Internship organized by the PI and Co-PI together with the Mathematics, Engineering, Science Achievement (MESA) program at USC will be offered to high-school students over the course of the project. Experience and knowledge gained by actively taking part in the project will attract this younger generation of scholars to the field of Biophysics and will provide them with strong scientific foundationsThe functional influence of cellular adhesion forces on the 3D plasticity of cav1 nano-domains and their spatial coupling to sub-membranous actin fibers and focal adhesions (FAs) will first be established by correlative 3D SR microscopy, robust spatial correlation analyses and modulation of adhesion geometry for micro-patterned cells. This will provide new understanding of the homeostatic PM functions of cav1 nano-domains as cells respond to specific mechanical constraints. The plasticity of cav1 nano-domains will then be quantitatively correlated with extra- and intracellular picoNewton forces developed along the PM at FAs by combining optical force sensor measurements, SR microscopy and cell micropatterning. This will shed new light on the role of cav1 as a mechano-transducer of extra/intracellular forces at the cell surface. Using quantum dot tracking of curvature- and non-curvature-coupled PM receptors together with 3D SR microscopy of cav1 nano-domains in live cells, the functional roles of cav1 nano-domains as local modulators of PM tension will then be established. This will reveal how they participate in adaptive coupling between local PM tension and cellular force generation on substrates. Finally, physical models describing the plasticity of cav1 nano-domains as a function of local membrane tension will be conceived and tested quantitatively to define some of the core physical principles that govern the adaptation of the cell PM to forces. Beyond offering new optical tools and original methodologies to study the function of PM nano-structures in cells, this work will bring novel insights into the physics of living systems by providing a mechanistic understanding of the physical principles that dictate PM plasticity and adaptation to forces at the nano-scale. This project is being jointly supported by the Physics of Living Systems program in the Division of Physics and the Cellular Dynamics and Function Program in the Division of Molecular and Cellular Biosciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

施加在细胞质膜（PM）上的机械力指导许多重要的细胞和组织过程，包括细胞粘附、迁移和侵袭。在这些过程中，膜弯曲蛋白小窝蛋白1（cav 1）和它在PM形成的不同类型的纳米级结构域正在成为哺乳动物细胞的关键机械转导枢纽和张力缓冲结构。然而，cav 1纳米级域的结构塑性和它们与PM的局部机械和张力状态的功能耦合仍然是谜。因此，细胞膜在纳米尺度上适应不同力的一些基本机制仍然不确定。该项目旨在了解cav 1纳米级域如何调节PM结构以响应机械提示，并定义管理PM可塑性，膜张力维持和细胞对力的适当反应的核心物理原理。该项目将通过多学科方法实施，该方法集成了超分辨率（SR）显微成像，光学力传感，细胞机械生物学，定量生物物理学和建模的生物材料工程。它将（i）定量定义细胞PM处cav 1纳米级域的纳米级组织，（ii）建立它们响应特定PM力的可塑性，（iii）确定它们如何局部调节PM张力，以及（iv）提供它们作为关键PM张力调制器的功能的物理模型。由于其原创性和多学科性质，该项目使其参与者参与高度跨学科的研究。它将为他们提供与细胞生物学，物理学和工程研究之间的当前融合相匹配的技能。拟议的活动还通过（一）成像探针、细胞培养和显微成像的实践实验，（二）外联项目和（三）教育活动，为传统上代表性不足的本科生和高中生提供一个融合现代技术和异花授粉科学的平台。经济上处于不利地位的本科生和高中生将通过南加州大学的既定项目和大洛杉矶地区的高中外展来招募。特别是，PI和Co-PI与南加州大学的数学，工程，科学成就（梅萨）计划一起组织的为期8周的暑期实习将在项目期间提供给高中生。通过积极参与该项目所获得的经验和知识将吸引年轻一代的学者进入生物物理学领域，并为他们提供强大的科学基础。细胞粘附力对cav 1纳米结构域的三维可塑性及其与膜下肌动蛋白纤维和粘着斑（FA）的空间耦合的功能影响将首先通过相关的三维SR显微镜建立，稳健的空间相关性分析和微图案化细胞的粘附几何形状的调制。这将为cav 1纳米结构域的稳态PM功能提供新的理解，因为细胞响应特定的机械约束。cav 1纳米域的可塑性，然后将定量相关的额外和细胞内的皮牛顿力开发沿着PM在FA通过结合光学力传感器测量，SR显微镜和细胞微图案。这将揭示cav 1作为细胞表面细胞内外力的机械换能器的作用。使用曲率和非曲率耦合PM受体的量子点跟踪以及活细胞中cav 1纳米域的3D SR显微镜，cav 1纳米域作为PM张力的局部调制器的功能作用将被建立。这将揭示它们如何参与局部PM张力和基底上细胞力产生之间的自适应耦合。最后，物理模型描述的可塑性cav 1纳米域作为局部膜张力的函数将被构思和定量测试，以定义一些核心的物理原则，管理的细胞PM的适应力。除了提供新的光学工具和原创方法来研究细胞中PM纳米结构的功能之外，这项工作还将通过提供对决定PM可塑性和适应力的物理原理的机械理解，为生命系统的物理学带来新的见解。纳米尺度。该项目由物理学系的生命系统物理学项目和分子与细胞生物科学系的细胞动力学和功能项目共同支持。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。