Impact of AICD on TCR Transduced T Cells for Adoptive Immunotherapy

过继性T细胞疗法（ACT）可以产生令人印象深刻的结果，但成功率很低。一般来说，成功与过继性转移T细胞的持久性有关。我们的研究结果表明，在ACT过程中发生的重复TCR刺激导致CDS效应细胞的激活诱导细胞死亡（AICD）。因此，AICD可能导致过继性转移T细胞的持久性差。最近，通过肿瘤特异性TCR（如高亲和力的TIL13S3i TCR）的转导，对自体T细胞进行遗传修饰已经成为可能。虽然对这些转基因细胞的生物学知之甚少，但我们发现它们也会经历AICD，因此，在表达TlL1383i TCR的载体中插入一个额外的保护基因是增强转基因T细胞持久性的可行翻译方法。因此，有必要了解导致AICD的信号转导途径。我们的初步数据确定JNK， p53和AIF是表达天然TCR的T细胞中AICD的介质。然而，表达天然TCR与转导TCR以及下游介质的T细胞之间的异同仍有待阐明。因此，我们将比较天然表达TlL1383i（源自h3T转基因模型）的T细胞和用TlL31S3i TCR转导的T细胞之间的AICD和抗肿瘤功效。我们观察到，ROS的产生是与AICD相关的一种常见现象，ROS清除剂的药理抑制可在不干扰T细胞活化的情况下保护细胞死亡，这使得氧化还原调节成为治疗干预的中心点。我们的假设是，从基因上增强TIL1383i TCR转导的T细胞的抗氧化能力，将导致对AICD的特异性和持续性保护，从而增加过继T细胞转移后的持久性、记忆池的发展和抗肿瘤功效。探讨反复刺激TCR后TCR转导T细胞发生AICD的机制。具体目标2。确定TCR抗氧化能力的增强对体外T细胞存活、功能和记忆发育的影响。具体目标3。评估过继转移TCR转导T细胞抗氧化能力的增强如何影响其体内持久性和记忆发展以及肿瘤消退。我们的方法可以直接影响正在进行的临床试验，其中转基因T细胞用于黑色素瘤的免疫治疗。

Adoptive T cell therapy (ACT) can lead to impressive results but the frequency of success is low. Generally success correlates with persistence of the adoptively transferred T cells. Our results indicate that repetitive TCR stimulation, which occurs during ACT leads to activation induced cell death (AICD) of CDS effector cells. Thus AICD may contribute to poor persistence of adoptively transferred T cells. Recently, it has become possible to genetically modify autologous T cells by transduction with a tumor-specific TCR such as the high affinity TIL13S3i TCR central to this application. While little is known about the biology of these genetically modified cells, we show that they also undergo AICD, Thus a feasible translational approach to enhance persistence of genetically modified T cells will be to insert an additional protective gene into the vector expressing the TlL1383i TCR. Therefore, it will be necessary to understand the signal transduction pathways leading to AICD. Our preliminary data identify JNK, p53, and AIF as mediators of AICD in T cells expressing a native TCR. However, similarities and differences between T cells expressing a native vs. a transduced TCR as well as downstream mediators and remain to be elucidated. We will therefore compare AICD and anti-tumor efficacy between T cells natively expressing the TlL1383i (derived from the h3T transgenic model) and T cells that have been transduced with the TlL31S3i TCR. We observed that ROS generation is a common phenomenon associated with AICD and that pharmacological inhibition with ROS scavenger results in protection from cell death without interfering with T cell activation, which places redox regulation at a central point for therapeutic intervention. It is our hypothesis is that genetically enhancing the antioxidant capacity of TIL1383i TCR transduced T cells will lead to specific and sustained protection from AICD thereby increasing persistence, memory pool development and anti-tumor efficacy following adoptive T cell transfer Our aims are: Specific Aim 1. To dissect the mechanisrh of AICD in TCR transduced T cells after repetitive TCR stimulation. Specific Aim 2. To determine how increasing the antioxidant capacity of TCR transduced T cells impacts, survival, function, and memory development in vitro. Specific Aim 3. To evaluate how increased antioxidant capacity of adoptively transferred TCR transduced T cells affects their persistence and memory development as well as tumor regression in vivo. Our approach could directly impact ongoing clinical trials, in which genetically modified T cells are used for immunotherapy of melanoma.