T cell migration and cardiovascular toxicity in immunotherapy

项目概要/摘要 T细胞免疫疗法正在成为一种有前途的癌症治疗选择，并已被证明在一系列领域有效。 恶性肿瘤然而，令人担忧的是， 过继转移体外活化T细胞到非靶组织部位可能易患心血管疾病， 损伤和全身炎症反应。关于心血管危害的传闻证据已经出现 大量数据表明与T细胞免疫治疗相关的细胞因子释放综合征的恶化。 我们进行这项研究，以解决关键的知识差距有关的分子机制，决定 过继转移的体外产生的T细胞的功能和命运，以及心血管毒性 这与静脉内转移后治疗性T细胞在非携带肿瘤的组织处的隔离有关。 通过我们初步研究的几条证据，我们提出自体T细胞经历了 在离体制造过程中显著的分子和细胞重编程。我们预测 内在变化对于T细胞的活化和扩增是重要的，但不能导致T细胞迁移 向靶肿瘤转移，从而增加毒性。我们发现β II-血影蛋白的减少 在体外T细胞活化过程中的表达导致细胞硬度降低，并且在细胞内表达的显著变化。 静脉内转移后自发T细胞迁移模式。此外，筛选关键的细胞内蛋白质 与改变的T细胞迁移相关，揭示了一种新的Rab 13介导的内体再分布模式 介导非特异性T细胞迁移。我们将：（1）确定心血管细胞毒性的原因 和与体外活化T细胞的非特异性迁移相关的细胞因子释放综合征，（2）确定 阻止向靶组织部位特异性迁移的分子机制，以及（3）测试我们是否 可以产生具有改善的组织特异性归巢特性和降低的心血管副作用的T细胞。 这些研究将结合联合收割机调节活化T细胞的新机制的差异扰动， 迁移，体内小鼠模型，最先进的活体多光子成像，高分辨率单细胞 测定和分析定义血管炎症反应，以了解潜在的严重风险， 过继性T细胞转移免疫疗法。我们还将探索新的替代方法， 抗癌功效和最小化T细胞免疫疗法的心血管风险。

PROJECT SUMMARY/ABSTRACT T cell immunotherapy is emerging as a promising cancer treatment option and has proven effective in a range of malignancy. However, a concern has been that prolong circulation and/or non-specific migration of the adoptively transferred in vitro activated T cells to non-target tissue sites might predispose to cardiovascular damages and systemic inflammatory responses. Anecdotal evidence of a cardiovascular hazard has emerged and abundant data point to exacerbation of cytokine release syndrome associated with T cell immunotherapy. We undertook this study to address critical knowledge gaps regarding the molecular mechanisms that determine the function and fate of the adoptively transferred in vitro-generated T cells, and cardiovascular toxicity associated with sequestration of the therapeutic T cells at non-tumor-bearing tissues after intravenous transfer. Through several lines of evidence from our preliminary study, we propose that autologous T cells undergo significant molecular and cellular reprogramming during ex-vivo manufacturing process. We predict that the intrinsic changes are important for the robust T cell activation and expansion, but fail to derive T cell migration toward the target tumor, and thus serve to increase toxicity. We discovered that a decrease in βII-spectrin expression during in vitro T cell activation results in decreased cell stiffness and a dramatic change in spontaneous T cell migration pattern upon intravenous transfer. Moreover, screening of a key intracellular protein associated with the altered T cell migration revealed a novel Rab13-mediated endosomal redistribution pattern that mediates the non-specific T cell migration. We will, (1) determine the causes of cardiovascular cytotoxicity and cytokine release syndrome associated with non-specific migration of in vitro activated T cells, (2) determine the molecular mechanisms that prevent specific migration toward the target tissue site, and (3) test whether we can generate T cells with an improved tissue-specific homing property and a reduced cardiovascular side-effects. These studies will combine differential perturbations of novel mechanisms that regulate activated T cell migration, in vivo mouse models, state of the art intravital multiphoton imaging, high-resolution singles cell assays, and analysis defining vascular inflammatory responses to understand a potentially serious risk of adoptively T cell transfer immunotherapy. We shall also explore novel alternative approaches that might promote the anti-cancer efficacy and minimize the cardiovascular risk of the T cell immunotherapy.