Understanding Metabolic and Epigenetic Cross‐ talk in Potent Anti‐ tumor T cells

摘要 最近的临床前研究表明，辅助性CD 4 + T辅助（Th）细胞和CD 8 + T辅助（Th）细胞的不同亚群可以通过免疫细胞化学方法检测到。 T细胞毒性（Tc）细胞有望在过继细胞治疗（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，并导致改善的肿瘤控制肿瘤， 与Th 1亚群（分泌IFNγ、IL 2、TNFα）相比。然而，与这些观察相反， 报道了与Tc 17细胞相比，Tc 1细胞表现出改善的肿瘤控制。T细胞中的这些差异 在抑制性肿瘤微环境中， 大部分肿瘤反应性T细胞获得FoxP 3+调节表型，变得功能失调， 经历导致肿瘤逆转的细胞死亡。因此，可以使T细胞成为免疫原性的离体编程条件是可行的。 稳定的表型，其不仅控制原发性肿瘤，而且导致抗肿瘤抗体的形成。 记忆力对ACT非常重要我们最近确定，编程条件 将Th 1细胞的“抗肿瘤效应功能”和Th 17细胞的“干细胞性”结合在一起上级 显示长期肿瘤控制的杂交Th 1/17细胞。重要的是，这些离体编程条件还 产生高效的杂交Tc 1/Tc 17细胞，并使人肿瘤浸润淋巴细胞（TIL）具有 细胞因子分泌能力增加。重要的是，杂交T细胞表现出更高水平的烟酰胺 腺嘌呤二核苷酸（NAD+），一种辅助因子，作为Sirtuins的底物，调节多种代谢 和表观遗传分子。我们假设，观察到的强大的长期肿瘤控制与杂交 Th 1/17细胞由整体再生的T细胞表型介导，这是由于高NAD+影响了Th 1/17细胞的分化。 事件的组合，包括翻译后修饰和表观遗传稳定性，导致 代谢适应表型。鉴于NAD+在包括能量在内的各种生物过程中的关键作用 代谢，衰老，钙稳态和表观基因组，我们提出了以下目标来测试上述 假设：目的1）为了确定是否代谢签名和细胞因子亚群的抗肿瘤表型， 杂交T1/17细胞可被翻译成编程肿瘤浸润淋巴细胞;目的2）确定是否 抗体介导的CD 38与PD 1抗体的组合抑制导致在肿瘤中的稳健抗肿瘤应答。 目的3）确定NAD+水平如何有助于不同的临床前体内和异种移植模型; 在T1/17杂合体和CD 38-KO T中保持稳健的抗肿瘤应答的代谢-表观遗传编程 细胞我们认为，这一建议，以确定中央机制，结果在上级抗肿瘤 通过杂交T1/17细胞的应答对于使这些离体编程条件适应于即刻免疫应答将是重要的。 在过继性T细胞免疫治疗中的翻译用途。

ABSTRACT Recent preclinical studies have shown that different subsets of both helper CD4+ T helper (Th) cells and CD8+ T cytotoxic (Tc) cells hold promise for clinical use in adoptive cell therapy (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 tumor reactive T cells acquire FoxP3+ regulatory phenotype, become dysfunctional or undergo cell death leading to tumor reversion. Thus, ex vivo programming conditions that can render T cells a stable phenotype, which not only controls primary tumors but also results in the formation of anti-tumor memory will be of immense importance for 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 cells exhibiting long-term tumor control. Importantly, these ex vivo programming conditions also generate highly effective hybrid Tc1/Tc17 cells and render human tumor infiltrating lymphocytes (TILs) with increased cytokine secreting ability. Importantly, the hybrid T cells exhibited higher levels of nicotinamide adenine dinucleotide (NAD+), a cofactor that serves as substrate for Sirtuins and regulates multiple metabolic and epigenetic molecules. We hypothesize that the robust long-term tumor control was observed with hybrid Th1/17 cells was mediated by an overall rejuvenated T cell phenotype due to high NAD+ that influenced a combination of events including post-translational modifications, and epigenetic stability that led to metabolically fit phenotype. Given the crucial role of NAD+ in a variety of biological processes including energy metabolism, aging, calcium homeostasis, and epigenome, we propose the following aims to test the above hypothesis: Aim 1) To determine if metabolic signature and anti-tumor phenotype of the cytokine subsets in hybrid T1/17 cells could be translated to program tumor infiltrating lymphocytes; Aim 2) To determine if antibody mediated combinatorial inhibition of CD38 with PD1 antibody leads to robust anti-tumor response in different preclinical in vivo and xenograft models; and Aim 3) To determine how NAD+ level contributes to metabolo-epigenetic programming that preserves robust anti-tumor response in T1/17 hybrid and CD38-KO T cells. We believe that this proposal to determine the central mechanism that results in superior anti-tumor response by hybrid T1/17 cells will be important to adapt these ex vivo programming conditions for immediate translational use in adoptive T-cell immunotherapy.