T cell migration and cardiovascular toxicity in immunotherapy

项目摘要/摘要 T细胞免疫疗法正在成为一种很有前途的癌症治疗选择，并已被证明在一系列治疗中有效 充满了恶意性。然而，一个令人关切的问题是，延长 体外过继转移活化T细胞到非靶组织部位可能易患心血管疾病 损害和全身炎症反应。关于心血管危险的轶事证据已经出现 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加剧。 我们进行了这项研究，以解决有关决定分子机制的关键知识空白 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 与静脉移植后治疗性T细胞在非荷瘤组织中的隔离有关。 通过我们初步研究的几条证据，我们认为自体T细胞经历了 在体外制造过程中显著的分子和细胞重新编程。我们预测， 固有的变化对于强大的T细胞激活和扩增很重要，但不能导致T细胞的迁移 对靶肿瘤的作用，从而增加毒性。我们发现βII-SPECTIN的减少 在体外T细胞激活过程中的表达导致细胞僵硬降低和显著变化 静脉转移时T细胞的自发迁移模式。此外，一种关键的胞内蛋白的筛选 与T细胞迁移相关的改变揭示了一种新的Rab13介导的内体重分布模式 介导了非特异性T细胞的迁移。我们将，(1)确定心血管细胞毒性的原因 和与体外激活的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 阻止特定迁移到目标组织部位的分子机制，以及(3)测试我们是否 可以产生T细胞，具有改善的组织特异性归巢特性，并减少心血管副作用。 这些研究将结合调节激活的T细胞的新机制的差异扰动 迁移，活体小鼠模型，最新的活体多光子成像，高分辨率单细胞 确定血管炎症反应的分析，以了解潜在的严重风险 过继T细胞转移免疫治疗。我们还将探索新的替代方法，以促进 T细胞免疫疗法的抗癌效果和最大限度降低心血管风险。

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.