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（汽车）转导。 最近，关于新抗原反应性T细胞或识别新突变抗原的T细胞的研究激增， 也表现出了承诺。虽然开展这些研究需要大量技术资源， 当使用效应T细胞反应性时，抗原丢失变体的出现也导致不太有效的肿瘤控制。 单个肿瘤抗原或靶分子。因此，在使用肿瘤浸润淋巴细胞方面有复苏 （TIL），其具有对多个肿瘤表位反应的内源性T细胞。虽然大多数研究 使用常规方法使用高剂量IL 2扩增TIL，一些最近的研究使用IL 15或IL 21 肿瘤控制得到改善。临床前研究还表明，辅助性CD 4+细胞和非辅助性CD 4+细胞的不同亚群， 辅助性T（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 具有分泌IL-17（Th 17）能力的辅助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，有报道称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细胞亚群反应控制 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，可以提供具有降低的表型的稳定表型的离体编程条件是可行的。 '可塑性'不仅控制原发性肿瘤，而且还导致抗肿瘤记忆的形成 在ACT中的重要性。我们最近已经确定， Th 1细胞的“抗肿瘤效应子功能”和Th 17细胞的“干细胞性”导致上级杂交Th 1/17（和 Tc/17）细胞表现出长期肿瘤控制。因此，在第一阶段的应用中，我们成功地测试了 将代谢增强特征编程为来自黑色素瘤和前列腺癌的TIL的可行性 患者对于II期应用，我们假设使用混合物的离体编程和扩增是可行的。 T1/17（Th 1/17和Tc 1/17）培养条件将导致稳健的抗肿瘤对照，即使具有更少的过继性 转移的临床级代谢增强的TIL（meTIL）。提出了以下具体目标， 建立和发展我们商业化方法：具体目标1：生产临床级杂交 来自黑素瘤和前列腺癌患者的METILs。具体目标2：确定临床级混合meTILs 在体内控制肿瘤生长方面上级常规TIL。我们认为，这项建议将有助于通过 用于产生未来可用于过继性T细胞的杂交meTILs的新的离体编程条件， 细胞免疫疗法临床试验。

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 tumor infiltrating lymphocytes (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) cell exhibiting long-term tumor control. Thus, in the Phase I application we successfully tested the feasibility of programming metabolically enhanced signature to TILs from melanoma and prostate cancer patients. For the Phase II application, we hypothesize that ex vivo programming and expansion using hybrid T1/17 (Th1/17 and Tc1/17) culture conditions will result in a robust anti-tumor control even with fewer adoptively transferred clinical grade metabolically enhanced TILs (meTILs). Following specific aims are proposed to establish and develop our approach for commercialization: Specific Aim 1: To generate clinical-grade hybrid meTILs from melanoma and prostate cancer patients. Specific Aim 2: To establish if clinical grade hybrid meTILs are superior to conventional TILs in controlling tumor growth in vivo. We believe that this proposal will help adopt the novel ex vivo programming conditions for generating hybrid meTILs that could be used future in adoptive T- cell immunotherapy clinical trials.