Materials and methods in quantifying cell mechanobiology

量化细胞力学生物学的材料和方法

• 批准号: RGPIN-2020-07169
• 负责人: Ehrlicher, Allen
• 金额: $ 2.4万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=741637
• 关键词: Materials; methods; quantifying; cell; mechanobiology

Spatiotemporal mechanical substrates and force microscopy advances to quantify cell contractility and viscoelasticity. Over the past decades, the forces and mechanical properties of biological systems have been recognized as an essential component across all scales of life. In particular, advents in elastic cell culture substrates have catalyzed a renaissance of cell biology, unlocking a previously unrecognized dimension in biological sciences. Understanding the physical mechanisms driving biological processes is a key challenge of the 21st century, and will resolve biological systems through an entirely new perspective of forces and mechanics, however, materials and methods to quantify these interactions are essential. Central to the active properties of most eukaryotic cells is that they are contractile, continuously exerting dynamic forces that pull on neighboring cells and their substrate. By culturing cells on deformable substrates, researchers have employed Traction Force Microscopy (TFM) to characterize the stiffness-dependent contractility of cells and the influence this contractility has on diverse biological processes from differentiation and proliferation to cancer metastasis. Bottlenecks of substrate stability, porosity, throughput, and technical complexity nevertheless have hampered adoption of these essential metrics in broader life-science applications. My lab has produced mechanically tunable silicone substrates for high-throughput contractile force measurements in diverse physiological and pathological contexts, and NSERC DG support has been critical in these advances. This DG proposal is focused on extending this frontier by developing (1) new materials and methodologies to quantify cell mechanics, with the goal of applying these to (2) quantify cell viscoelasticity. To do so, we will create silicone-based cell substrates that are a) elastically patterned in space; b) elastically switchable in time; c) adhesively patterned to direct single and collective cell structure dimensionality; and d) hydrogel substrates that locally contract to stretch cells and probe their mechanical response. My lab will build on our expertise with TFM to both simplify and enhance quantification of real-time cell contractility and mechanical properties. We will selectively modify key intracellular and intercellular proteins and quantify their contributions to contractile forces and cellular viscoelasticity. This DG program is built on a firm foundation of my lab's expertise in silicone substrate mechanics and cell force measurements, and well-supported by extensive microscopy and mechanical-characterization instrumentation. My DG program will drive a materials-based quantification of cell contractility and viscoelasticity. Due to the broad and fundamental utility of the innovations proposed here, I anticipate far-reaching impact in biological physics, quantitative biology, materials science, and experimental medicine.

时空机械底物和迫使显微镜的前进以量化细胞收缩性和粘弹性。在过去的几十年中，生物系统的力和机械性能已被认为是所有范围的重要组成部分。特别是，弹性细胞培养底物的优势催化了细胞生物学的复兴，解锁了先前未识别的生物科学维度。了解驱动生物学过程的物理机制是21世纪的关键挑战，它将通过全新的力量和力学观点来解决生物系统，但是，量化这些相互作用的材料和方法至关重要。 大多数真核细胞的活性特性的核心是它们是收缩的，不断地施加动态力，这些动态力吸引了相邻细胞及其底物。通过在可变形底物上培养细胞，研究人员采用了牵引力显微镜（TFM）来表征细胞的僵硬依赖性收缩力，并且该收缩力对从分化和增殖到癌症转移的多种生物学过程产生了影响。但是，底物稳定性，孔隙率，吞吐量和技术复杂性的瓶颈仍阻碍了在更广泛的生活科学应用中对这些基本指标的采用。我的实验室已经在不同的生理和病理环境中生产了机械可调的有机硅底物，用于高通量收缩力测量，而NSERC DG支持在这些进展中至关重要。 该DG提案的重点是通过开发（1）新材料和方法来量化细胞力学，以将其应用于（2）量化细胞粘弹性，以扩展该领域。为此，我们将创建基于硅酮的细胞底物，该基材是a）在空间中弹性图案的； b）及时可弹性切换； c）具有粘附的图案化，以指导单一和集体的细胞结构维度； d）局部收缩以拉伸细胞和探测其机械响应的水凝胶底物。我的实验室将基于我们的TFM专业知识，以简化和增强实时细胞收缩性和机械性能的量化。我们将有选择地修改关键细胞内和细胞间蛋白，并量化它们对收缩力和细胞粘弹性的贡献。该DG程序建立在我实验室在硅胶底物力学和细胞力测量方面的专业知识的基础上，并通过广泛的显微镜和机械性特征仪器良好支持。我的DG程序将推动基于材料的细胞收缩性和粘弹性定量。由于这里提出的创新的广泛而基本的实用性，我预计对生物物理学，定量生物学，材料科学和实验医学的影响深远。