Immunoengineering cellobiose as a fuel source for T cells

摘要 由于肿瘤的贪婪性质，实体肿瘤的微环境中的葡萄糖水平较低。 新陈代谢，阻碍肿瘤浸润性T细胞的功能，否则可能控制肿瘤的生长。T 细胞需要葡萄糖来维持能量、细胞因子的产生、增殖和细胞毒性。注入更多的 葡萄糖注入患者并不是一个可行的解决方案，因为它只会喂养肿瘤，并进一步饥饿T细胞。领养 细胞疗法(CAR-T细胞)在实体肿瘤中常常失败，因为肿瘤造成的代谢障碍。 纤维二糖是一种葡萄糖聚合物，在植物物质中以纤维素的形式大量存在，具有潜在的 作为一种碳和能源。然而，哺乳动物细胞不能分解纤维二糖。我们的预赛 数据显示，将两种蛋白质改造成T细胞，可以让它们利用纤维二糖。因此，我们已经有了 一种工作系统，旨在改造T细胞，使其拥有对抗肿瘤的唯一葡萄糖来源。我们展示了 这种葡萄糖来源对肿瘤来说是完全无法获得的。 我们的长期目标是将这种能力转化为用于癌症免疫治疗的T细胞 (例如，CAR-T细胞)。我们的团队包括加州大学洛杉矶分校的T细胞免疫学家，并得到免疫领域的专家同事的协助 新陈代谢和癌症免疫疗法。在这个R21提案中，我们将在目标1中将其描述为纤维二糖 水解成葡萄糖，并将其碳追踪到各种代谢和生物合成途径中，并提供 能量。我们通过最先进的代谢组学测试细胞利用纤维二糖衍生葡萄糖的能力 293个细胞系和原代T细胞。在目标2中，我们测试毒性和药代动力学以准备治疗黑色素瘤。 在老鼠身上做实验。我们在一个概念验证实验中测试了T细胞对抗小鼠黑色素瘤的能力， 使用转导的PMEL TCR转基因基因和针对黑色素瘤的CAR-T细胞。 通过将纤维二糖代谢添加到工程T细胞中，我们提供了一种新的燃料来源和协同方法 这大大加强了癌症免疫疗法。

Abstract Glucose levels are low in the microenvironment of solid tumors due to the voracious nature of tumor metabolism, impeding the function of tumor-infiltrating T cells that might otherwise control tumor growth. T cells require glucose for energetics, cytokine production, proliferation, and cytotoxicity. Infusing additional glucose into patients is not a viable solution as it would feed only the tumor and further starve T cells. Adoptive cell therapies (CAR-T cells) often fail in solid tumors because of this metabolic hurdle created by tumors. Cellobiose, a polymer of glucose found abundantly in plant matter in the form of cellulose, has the potential to serve as a carbon and energy source. However, mammalian cells cannot catabolize cellobiose. Our preliminary data show that engineering two proteins into T cells allows them to make use of cellobiose. Thus, we already have a working system set up to engineer T cells to have an exclusive source of glucose to fight tumors. We showed this glucose source is completely inaccessible to tumors. Our long-term goal is to translate this capability to T cells that are engineered for cancer immunotherapy (e.g., CAR-T cells). Our team includes T-cell immunologists at UCLA assisted by expert colleagues in immune metabolism and cancer immunotherapies. In this R21 proposal, we will in Aim 1, characterize as cellobiose is hydrolyzed to glucose, and trace its carbons into various metabolic and biosynthetic pathways and provide energy. We test the capacity of cells to use cellobiose-derived glucose through state-of-the-art metabolomics in 293 cell lines and primary T cells. In Aim 2, we test toxicity and pharmacokinetics to prepare for melanoma experiments in mice. We test in a proof-of-concept experiment the ability of T cells to fight mouse melanomas, using transduced pmel TCR transgenics and CAR-T cells that target melanoma. By adding cellobiose metabolism to engineered T cells, we offer a new fuel source and a synergistic approach that significantly potentiates cancer immunotherapies.