Role of lipid metabolism in CD8+ T cell ferroptosis

脂质代谢在 CD8 T 细胞铁死亡中的作用

• 批准号: 10792062
• 负责人: Qing Yi
• 金额: $ 48.45万
• 依托单位: METHODIST HOSPITAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10792062
• 关键词: Adoptive Cell Transfers; Beds; Blood; Bone Marrow; CD36 gene; CD8-Positive T-Lymphocytes; CD8B1 gene; Cancer Patient; Cell physiology; Cells; Cellular Metabolic Process; Cessation of life; Clinical; Coenzyme A; Cytotoxic T-Lymphocytes; Data; Enzymes; Fatty Acids; Functional disorder; Gene Expression; Genes; Immunotherapy; Impairment; In complete remission; Individual; Infiltration; Interferon Type II; Lipid Peroxidation; Malignant Neoplasms; Mediating; Melanoma Cell; Memory; Metabolic; Methods; Modeling; Multiple Myeloma; Mus; Oxidoreductase; Patients; Polyunsaturated Fatty Acids; Production; Prognosis; Publications; Reporting; Role; T-Lymphocyte; TNF gene; Testing; Treatment Efficacy; Tumor Burden; cancer immunotherapy; cancer infiltrating T cells; comparison control; cytokine; cytotoxic; cytotoxic CD8 T cells; effector T cell; fatty acid oxidation; immune checkpoint blockade; improved; individual patient; knock-down; lipid metabolism; melanoma; new therapeutic target; novel; novel therapeutic intervention; oxidation; programs; response; single-cell RNA sequencing; success; transcriptome sequencing; tumor; tumor microenvironment; uptake

PROJECT SUMMARY Recently, we made a novel and exciting discovery that tumors or the tumor microenvironment (TME) cause T- cell dysfunction and death by inducing ferroptosis in T cells. We analyzed the sc-RNA-seq data of tumor- infiltrating T cells from melanoma patients and discovered that tumor-infiltrating CD8+ T cells had significantly increased expressions of genes associated with lipid peroxidation and ferroptosis compared to blood CD8+ T cells from healthy individuals. More importantly, our unpublished studies further revealed that among different CD8+ T cell subpopulations, effector memory (TEM) and terminally differentiated effector (TTE) CD8+ T cells are more sensitive to tumor-induced ferroptosis. We examined tumor-infiltrating CD8+ T cells from the bone marrow (BM; tumor bed) of patients with multiple myeloma. By separating the T cells into naïve, TTE or TEM cells based on their expression of CCR7 and CD45RA, we found that TEM and TTE CD8+ T cells expressed higher levels of lipid peroxidation- and ferroptosis-associated genes and were more sensitive to tumor-induced ferroptosis compared to naïve CD8+ T cells although they expressed similar levels of CD36. Similarly, in mouse melanoma and MM models, increased ferroptosis mainly occurred in tumor-infiltrating CD8+ TEM and TTE cells but not in naïve CD8+ T cells from mice with large tumor burdens compared to those with small tumor burdens. Our ex vivo studies confirmed that CD8+ TEM and TTE cells were more sensitive to tumor- or FA- induced ferroptosis than naïve T cells and their production of cytotoxic cytokines such as IFNγ and TNFα was inhibited. To elucidate the underlying mechanisms, RNA-seq was used and showed that CD8+ TEM and TTE cells expressed a significantly lower level of 2,4-dienoyl-CoA reductase 1 (DECR1), a rate-limiting enzyme for polyunsaturated fatty acid (PUFA) β-oxidation, compared to naïve CD8+ T cells. Knockdown (KD) of DECR1 in naïve CD8+ T cells resulted in an increased PUFA expression and peroxisomal dysfunction and sensitized them to tumor- or FA-induced ferroptosis than control T cells. Based on these novel findings, we hypothesize that CD8+ TEM and TTE cells, due to uptake of more FAs and reduced expression of DECR1 and PUFA oxidation, are sensitive to tumor/TME-induced ferroptosis, and inhibiting TEM and TTE cell ferroptosis may effectively enhance the therapeutic efficacy of immunotherapy in cancer patients. Aim 1 will determine the mechanism underlying tumor- or FA-induced ferroptosis in CD8+ TEM and TTE cells in TME. Aim 2 will elucidate the role and mechanisms of tumor and TME accumulation of FAs and induction of lipid peroxidation in CD8+ TEM and TTE T cells and Aim 3 will inhibit CD8+ TEM and TTE cell ferroptosis to enhance the efficacy of cancer immunotherapies. Accomplishing these aims will provide us with in-depth understanding of the mechanisms underlying how tumors and the TME induce T cell lipid peroxidation and how ferroptosis mediates T cell metabolic malfunction and death. Understanding these mechanisms is extremely important and will greatly assist us in developing novel therapeutic approaches to target T cell lipid metabolism and TME to significantly improve the efficacy of cancer immunotherapy.

项目概要 最近，我们有了一个令人兴奋的新发现，即肿瘤或肿瘤微环境 (TME) 会导致 T- 通过诱导 T 细胞铁死亡而导致细胞功能障碍和死亡。我们分析了肿瘤的 sc-RNA-seq 数据 研究人员对来自黑色素瘤患者的浸润性 T 细胞进行了研究，发现肿瘤浸润性 CD8+ T 细胞显着 与血液 CD8+ T 相比，与脂质过氧化和铁死亡相关的基因表达增加 来自健康个体的细胞。更重要的是，我们未发表的研究进一步表明，在不同的 CD8+ T 细胞亚群、效应记忆 (TEM) 和终末分化效应 (TTE) CD8+ T 细胞 对肿瘤引起的铁死亡更敏感。我们检查了来自骨骼的肿瘤浸润性 CD8+ T 细胞 多发性骨髓瘤患者的骨髓（BM；瘤床）。通过将 T 细胞分为 naïve、TTE 或 TEM 根据细胞中 CCR7 和 CD45RA 的表达，我们发现 TEM 和 TTE CD8+ T 细胞表达 脂质过氧化和铁死亡相关基因水平较高，并且对肿瘤诱导的疾病更敏感 与幼稚 CD8+ T 细胞相比，铁死亡，尽管它们表达的 CD36 水平相似。同样，在 小鼠黑色素瘤和 MM 模型中，铁死亡增加主要发生在肿瘤浸润性 CD8+ TEM 和 TTE 细胞，但不存在于来自肿瘤负荷较大的小鼠（与肿瘤较小的小鼠相比）的初始 CD8+ T 细胞中 负担。我们的离体研究证实 CD8+ TEM 和 TTE 细胞对肿瘤或 FA- 更敏感 与幼稚 T 细胞相比，它们诱导铁死亡，并且它们产生细胞毒性细胞因子（例如 IFNγ 和 TNFα） 抑制。为了阐明潜在机制，使用 RNA-seq 并显示 CD8+ TEM 和 TTE 细胞表达的 2,4-二烯酰辅酶 A 还原酶 1 (DECR1) 水平显着降低，这是一种限速酶。 与初始 CD8+ T 细胞相比，多不饱和脂肪酸 (PUFA) β-氧化。 DECR1 的敲低 (KD) 初始 CD8+ T 细胞导致 PUFA 表达增加和过氧化物酶体功能障碍并致敏 与对照 T 细胞相比，它们更容易导致肿瘤或 FA 诱导的铁死亡。基于这些新发现，我们假设 CD8+ TEM 和 TTE 细胞，由于摄取更多 FA 以及 DECR1 和 PUFA 表达减少 氧化，对肿瘤/TME 诱导的铁死亡敏感，抑制 TEM 和 TTE 细胞铁死亡可能 有效增强癌症患者免疫治疗的治疗效果。目标 1 将确定 TME 中 CD8+ TEM 和 TTE 细胞中肿瘤或 FA 诱导的铁死亡的机制。目标 2 将阐明 肿瘤和 TME 中 FA 积累和诱导 CD8+ 脂质过氧化的作用和机制 TEM和TTE T细胞和Aim 3将抑制CD8+ TEM和TTE细胞铁死亡以增强癌症功效 免疫疗法。实现这些目标将使我们对机制有深入的了解 肿瘤和 TME 如何诱导 T 细胞脂质过氧化以及铁死亡如何介导 T 细胞的基础 代谢功能障碍和死亡。了解这些机制极其重要，并且将极大地促进 协助我们开发针对 T 细胞脂质代谢和 TME 的新型治疗方法，以显着改善 提高癌症免疫治疗的疗效。