QUANTIFYING EXTRACELLULAR MATRIX MECHANICAL PROPERTIES DUE TO MT1-MMP ACTIVITY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们将使用提出的LAMMP空间调制微束(SMM)系统来测量细胞外基质(ECM)对MT1-MMP表达细胞的响应中的机械变化。在我们的第一个目标中，我们将测量细胞外基质(ECM)蛋白与MT1-MMP表达细胞表面的膜受体之间的粘附力。包覆有特定ECM蛋白的微珠被固定在光学力夹具中，以测量粘附力。一个基于FRET的细胞表面MT1-基质金属蛋白酶活性传感器将监测MT1-基质金属蛋白酶活性水平，这将在时间上与黏附强度相关。在我们的第二个目标中，表达MT1-MMP的细胞将被培养在含有不同类型的ECM蛋白的ECM蛋白水凝胶中。通过拉动和测量嵌入在水凝胶中的多个微珠的位移，我们将跟踪不同表达水平的细胞对化学刺激的反应，凝胶近端和远端的机械性能的变化。这一合作将为膜结合的MT1-MMP消化ECM的动力学提供新的见解。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We will use the proposed LAMMP Spatially Modulated Microbeams (SMM) system to measure mechanical changes in the extra cellular matrix (ECM) in response to MT1-MMP expressing cells. In our first aim, adhesion forces between extracellular matrix (ECM) proteins and membrane receptors on the surface of MT1-MMP expressing cells will be measured. Microbeads coated with specific ECM proteins we be held in an optical force clamp to measure adhesion forces. A FRET based sensor of MT1-MMP activity on the cell surface will monitor MT1-MMP activity levels, which will be correlated in time to adhesion strength. In our second aim, MT1-MMP expressing cells will be cultured within ECM protein hydrogels containing various mixtures of ECM protein types. By pulling and measuring displacements of multiple microbeads embedded within the hydrogel, we will track changes in mechanical properties of the gel proximal and distal to cells with varying expression levels in response to chemical stimulation. This collaboration will offer new insights into the kinetics of ECM digestion by membrane-bound MT1-MMP.