Immunoengineering cellobiose as a fuel source for T cells

摘要 由于肿瘤的贪婪性质，在实体瘤的微环境中葡萄糖水平较低 这可能会导致肿瘤细胞的新陈代谢，阻碍肿瘤浸润性T细胞的功能，否则可能会控制肿瘤生长。不 细胞需要葡萄糖用于能量、细胞因子产生、增殖和细胞毒性。注入额外的 将葡萄糖注入患者体内不是可行的解决方案，因为它只会喂养肿瘤并进一步饿死T细胞。养父 细胞疗法（CAR-T细胞）在实体瘤中往往失败，因为肿瘤产生了这种代谢障碍。 纤维二糖是一种葡萄糖聚合物，以纤维素的形式大量存在于植物物质中， 作为一种碳和能源。然而，哺乳动物细胞不能分解代谢纤维二糖。我们的初步 数据显示，将两种蛋白质工程改造到T细胞中，使它们能够利用纤维二糖。因此，我们已经有 这是一个工作系统，可以设计T细胞，使其拥有对抗肿瘤的唯一葡萄糖来源。我们展示 这种葡萄糖源是肿瘤完全无法接近的。 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 (e.g., 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， 代谢和癌症免疫疗法。在该R21提案中，我们将在目标1中，将纤维二糖表征为 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， 能源我们通过最先进的代谢组学测试细胞使用纤维二糖衍生的葡萄糖的能力， 293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，以准备黑色素瘤 小鼠实验我们在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转基因和靶向黑素瘤的CAR-T细胞。 通过将纤维二糖代谢添加到工程化T细胞中，我们提供了一种新的燃料来源和协同方法 能显著增强癌症免疫治疗的药物。

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.