T Cell Mechanosensing of Microscale Fibers

摘要 机械力在T细胞和抗原提呈细胞之间的通讯中起着关键作用 (APCS)。我们课题组之前的研究表明，T细胞可以感知并对 TCR/CD3和CD28的活化底物的机械刚性，本质上是一种 工程化APC。这一发现导致了改进人类T细胞生产的新系统 免疫疗法。特别是，基于软弹性体纤维的系统每轮产生更多的细胞 比刚性对应物扩增，也挽救了正在接受治疗的个体的细胞扩增 癌症。然而，这些先前的实验也表明，潜在的机械感觉反应是 不仅取决于材料的体积模数，而且还取决于其结构，其规模可与之媲美，甚至可以媲美 比附着的细胞大。这一探索性/开发性项目的总体目的是 确定该网状平台的哪些特征，例如纤维直径和/或跨度长度会影响T细胞 机械传感。这将通过两个相辅相成的具体目标来实现。在第一，一套战略性的 独立地改变感兴趣的多个参数的光纤几何形状将通过进一步 发展电纺和微细加工弹性体。多种T细胞免疫应答的比较 这些纤维将被用来分离和量化每个参数的影响。第二个具体目标 聚焦于附着的T细胞，描述导致力产生的内部组件的特征。一起， 这些研究将解决目前关于T细胞如何机械地相互作用的知识空白 地形复杂的底物，提供了对这些细胞如何识别和攻击目标的洞察 改善新兴的细胞免疫治疗市场的细胞和策略。

Abstract Mechanical forces play critical roles in communication between T cells and Antigen Presenting Cells (APCs). Previous studies from our research group demonstrated that T cells can sense and respond to the mechanical rigidity of an activating substrate presenting ligands to TCR/CD3 and CD28, essentially an engineered APC. This discovery led to new systems for improving production of human T cells for cellular immunotherapy. In particular, a system based on soft elastomer fibers produced more cells per round of expansion than a rigid counterpart, and also rescued expansion of cells from individuals being treated for cancer. However, these previous experiments also suggested that the underlying mechanosensing response is dependent on not only the bulk modulus of the material but also its structure at scales comparable to or even larger than that of the attached cells. The overall purpose of this exploratory / developmental project is to identify what features of this mesh platform, such as fiber diameter and/or span length influence T cell mechanosensing. This will be approached in two complementary Specific Aims. In the first, a strategic set of fiber geometries that independently vary the multiple parameters of interest will be created by further developing electrospinning and microfabrication of elastomers. Comparison of multiple T cell responses to these fibers will be used to separate and quantify the effects of each parameter. The second Specific Aim focuses on the adherent T cell, characterizing the internal assemblies that lead to force generation. Together, these studies will address the current gap in knowledge of how T cells mechanically interact with topographically complex substrates, providing both insights into how these cells recognize and attack target cells and strategies to improve the emerging cellular immunotherapy market.