IMPACT OF AICD ON TCR TRANSDUCED T CELLS FOR ADOPTIVE IMMUNOTHERAPY

过继T细胞治疗(ACT)可以取得令人印象深刻的结果，但成功率很低。一般来说，成功与过继转移的T细胞的持久性有关。我们的结果表明，在ACT期间发生的重复TCR刺激会导致CDS效应细胞的激活诱导细胞死亡(AICD)。因此，AICD可能是过继转移的T细胞持久性差的原因之一。最近，通过转导肿瘤特异性TCR来修饰自体T细胞已经成为可能，例如对这一应用至关重要的高亲和力TIL13S3i TCR。虽然对这些转基因细胞的生物学知之甚少，但我们发现它们也经历了AICD，因此一个可行的翻译方法将是在表达TlL1383i TCR的载体中插入额外的保护性基因来提高转基因T细胞的持久性。因此，有必要了解导致AICD的信号转导通路。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的介体。然而，表达天然TCR和转导TCR以及下游介体的T细胞之间的异同仍有待阐明。因此，我们将比较天然表达Tl1383i的T细胞(来自H3T转基因模型)和转导了Tl31S3i TCR的T细胞之间的AICD和抗肿瘤效果。我们观察到，ROS的产生是AICD的常见现象，用ROS清除剂进行药物抑制可以在不干扰T细胞激活的情况下保护细胞免于死亡，这将氧化还原调节置于治疗干预的中心点。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的特异和持续的保护，从而增加过继T细胞转移后的持久性、记忆池发育和抗肿瘤效果。我们的目的是：具体目的：1.分析重复TCR刺激后转导TCR的T细胞发生AICD的机制。具体目的2.确定增加TCR转导的T细胞的抗氧化能力对体外存活、功能和记忆发育的影响。具体目的3.评价过继转移的TCR转导的T细胞的抗氧化能力增强对其持久性、记忆力发展以及体内肿瘤消退的影响。我们的方法可能直接影响正在进行的临床试验，在这些试验中，转基因T细胞被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 ofthe 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 mechanism 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.