Biomimetic Matrix for Ex Vivo and In Vivo Activation of T Cells

项目摘要 本研究的目的是设计一种人工T细胞刺激基质（aTM）， 以及细胞特异性生化和生物物理线索来控制表型并改善T细胞的功能特征 仿生背景，捕获淋巴结（LN）的关键生化和生物物理特征。LN提供了 关键的微环境，其协调抗原特异性信号的呈现格式和动力学， 精确和受控的方式，导致T细胞活化，扩增和成熟。目前可用的T细胞 然而，携带抗原呈递复合物和共刺激因子的刺激基质缺乏 LN中T细胞刺激微环境固有的足够的支持线索。我们将设计和 表征整合了三种关键T细胞刺激信号的aTM：抗原特异性（信号1）、共刺激性（信号2）、免疫调节性（信号3）和免疫调节性（信号4）。 细胞因子（信号2）和细胞因子（信号3），连同细胞外基质（ECM）分子和可调生物物理活性分子， 受抗原呈递细胞（APC）和LN的性质启发的性质。我们将检验这个假设 在T细胞刺激过程中，在这种仿生环境中共同呈递APC信号提示将导致功能性 具有受控表型特征和效力的T细胞。揭示了所有信号线索如何协同作用， 活化和活化T细胞。在具体目标1中，我们将确定aTM的关键矩阵属性， 使用由透明质酸（HA）水凝胶和HA-HA复合物（NHC）制备的aTM的CD 8 + T细胞活化 矩阵在具体目标2中，我们将定义最佳信号提示及其呈现配置， aTM上的CD 4 + T细胞活化和极化;并证明aTM刺激的CD 4 + T细胞活化和极化的持续性和功能。 过继转移的抗原特异性CD 4 + T细胞。在特定目标3中，我们将使用aTM微粒来共刺激 CD 8+和CD 4 + T细胞的联合治疗，并评估CD 4+和CD 8 + T细胞在 癌症免疫治疗小鼠模型。如果成功的话，这项研究将产生第一组工程化的aTM， - 提供对抗原特异性CD 8+和CD 4 + T细胞的强刺激，这有望改善治疗结果; 进一步丰富了我们对信号线索在刺激，极化， 和T细胞的激活。

PROJECT SUMMARY The objective of this study is to engineer an artificial T cell-stimulating matrix (aTM) that presents antigen-specific and cell-specific biochemical and biophysical cues to control phenotype and improve functional profiles of T cells in a biomimetic context that captures key biochemical and biophysical features of the lymph node (LN). LN provides the critical microenvironment that orchestrates the presentation format and dynamics of the antigen-specific signals in a precise and controlled manner that leads to T cell activation, expansion, and maturation. Currently available T cell stimulating matrices while carrying the antigen-presentation complexes and co-stimulating cues, however, lacks adequate supporting cues inherent to the T cell stimulating microenvironment in the LN. We will design and characterize an aTM that integrates the three key T-cell stimulating signals: antigen-specific (Signal 1), co-stimulatory (Signal 2), and cytokines (Signal 3), together with extracellular matrix (ECM) molecules and tunable biophysical properties inspired by the properties of both antigen-presenting cells (APCs) and the LN. We will test the hypothesis that co-presenting the APC signal cues in such a biomimetic context during T cell stimulation will result in functional T cells with controlled phenotypic profiles and potency. and mechanism revealing how all signaling cues synergize to active and polarize T cells. In Specific Aim 1, we will determine key matrix properties of the aTM that mediate robust CD8+ T cell activation using aTMs prepared from hyaluronic acid (HA) hydrogel and HA-nanofiber composite (NHC) matrix. In Specific Aim 2, we will define optimal signaling cues and their presentation configuration that influence CD4+ T cell activation and polarization on aTM; and demonstrate persistence and functions of aTM-stimulated and adoptively transferred antigen-specific CD4+ T cells. In Specific Aim 3, we will use aTM microparticles to co-stimulate CD8+ and CD4+ T cells and evaluate therapeutic benefits of a combination treatment with CD4+ and CD8+ T cells in a cancer immunotherapy mouse model. If successful, this study will result in the first set of engineered aTM that delivers robust stimulation of antigen-specific CD8+ and CD4+ T cells that promise improved therapeutic outcomes; and further enrich our understandings of design principles and mechanism of signaling cues in stimulation, polarization, and activation of T cells.