Programming Metabolically Fit TILs for Immunotherapy

摘要 分子生物学和基因工程的进步导致了修饰T细胞的设计和使用 识别肿瘤以在过继细胞转移（ACT）至患者时实现显著的肿瘤控制。这些T 细胞用肿瘤抗原反应性T细胞受体（TCR）或嵌合抗原受体转导 （汽车）。最近，关于新抗原反应性T细胞或识别新突变的T细胞的研究激增， 抗原也显示出希望。虽然这些研究的实施需要大量的技术支持， 由于缺乏足够的资源，抗原丢失变体的出现也导致使用时肿瘤控制效果较差。 对单一肿瘤抗原或靶分子反应的效应T细胞。因此，在使用TILs方面有复苏， 其具有对多种肿瘤表位反应的内源性T细胞。虽然大多数研究都使用 使用高剂量IL 2扩增TIL的常规方法，一些最近使用IL 15或IL 21的研究显示， 改善肿瘤控制。临床前研究还表明，辅助性CD 4 + T细胞的不同亚群， (Th)细胞和CD 8 + T细胞毒性（Tc）细胞有望在ACT方案中临床使用。重要的是，T辅助细胞 具有分泌IL-17（Th 17）能力的细胞亚群已显示具有干细胞样表型， 这归因于它们的长期持久性，并导致与Th 1相比改善的肿瘤控制肿瘤。 亚群（分泌IFNγ、IL 2、TNFα）。然而，与这些观察相反，有报道说，Tc 1 细胞与Tc 17细胞相比显示出改善的肿瘤控制。这些差异在T细胞亚群响应 在抑制性肿瘤微环境中， 这些Th或Tc亚群获得FoxP 3+调节表型，变得功能障碍或经历细胞死亡 导致肿瘤逆转。因此，可以提供具有以下特征的稳定表型的离体编程条件： 降低“可塑性”，不仅控制原发性肿瘤，而且还导致形成抗肿瘤记忆， 这在ACT中非常重要。我们最近已经确定， Th 1细胞的“抗肿瘤效应功能”和Th 17细胞的“干细胞性”一起导致上级杂交Th 1/17 (and Tc/17）细胞表现出长期肿瘤控制。因此，我们假设离体扩增和 将TIL编程为杂交T1/17（Th 1/17和Tc 1/17）表型将导致稳健的抗肿瘤控制， 具有更少的过继转移细胞。为建立和发展我们的 商业化的方法：具体目标1：确定是否可以将人黑色素瘤肿瘤源性TIL 离体编程为有效的抗肿瘤杂合体T1/17表型。具体目标2：确定混合TILs是否 在体内控制黑素瘤肿瘤生长方面上级常规TIL。我们认为，这项建议 将有助于采用新的离体编程条件来产生强大的抗肿瘤TIL， 未来将用于过继性T细胞免疫治疗临床试验。

ABSTRACT Advances in molecular biology and genetic engineering have led to the design and use of modified T cells recognize tumors to achieve significant tumor control upon adoptive cell transfer (ACT) to patients. These T cells are either transduced with tumor antigen reactive T cell receptors (TCR), or chimeric antigen receptors (CARs). Recently, a surge in studies with neo-antigen reactive T cells or T cells recognizing novel mutated antigens has also shown promise. While the implementation of these studies requires significant technical resources, the appearance of antigen loss variants also leads to less effective tumor control when using effector T cells reactive to single tumor antigen or target molecule. Thus, there is a resurgence in using TILs, which have endogenous T cells reactive to multiple tumor epitopes, for ACT. While most studies have used the conventional approach to expand TILs using high dose IL2, some recent studies using IL15 or IL21 showed improved tumor control. Preclinical studies have also shown that different subsets of both helper CD4+ T helper (Th) cells and CD8+ T cytotoxic (Tc) cells hold promise for clinical use in ACT protocols. Importantly, T helper cell subsets with the ability to secrete IL-17 (Th17) have been shown to possess stem cell like phenotype that attributes to their long-term persistence and leads to improved tumor control tumors as compared to the Th1 subsets (that secrete IFNγ, IL2, TNFα). However, contrary to these observations there are reports that Tc1 cells exhibit improved tumor control as compared to Tc17 cells. These differences in T cell subsets response to control tumors, is compounded by the fact that in the suppressive tumor microenvironment a large fraction of these Th or Tc subsets acquire FoxP3+ regulatory phenotype, become dysfunctional or undergo cell death leading to tumor reversion. Thus, ex vivo programming conditions that can render a stable phenotype with reduced `plasticity' and not only controls primary tumors, but also results in formation of anti-tumor memory will be of immense importance in ACT. We have recently established that programming conditions that bring together `anti-tumor effector function' of Th1 cells and `stemness' of Th17 cells lead to a superior hybrid Th1/17 (and Tc/17) cells exhibiting long-term tumor control. Thus, we hypothesize that ex vivo expansion and programming of TILs to hybrid T1/17 (Th1/17 and Tc1/17) phenotype will lead to robust anti-tumor control even with fewer adoptively transferred cells. Following specific aims are proposed to establish and develop our approach for commercialization: Specific Aim 1: To determine if human melanoma tumor derived TILs could be ex vivo programmed to potent anti-tumor hybrid T1/17 phenotype. Specific Aim 2: To establish if hybrid TILs are superior to conventional TILs in controlling melanoma tumor growth in vivo. We believe that this proposal will help adopt the novel ex vivo programming conditions for generating robust anti-tumor TILs that could be used future in adoptive T-cell immunotherapy clinical trials.