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 More importantly, our unpublished studies further revealed that among different CD8+ T cell subpopulations, effector memory (TEM) and terminally differentiated effector (TTE) CD8+ T cells are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.

项目摘要 最近，我们有了一个新的令人兴奋的发现，肿瘤或肿瘤微环境（TME）导致T- 细胞功能障碍和死亡。我们分析了肿瘤的sc-RNA-seq数据， 黑色素瘤患者的浸润性T细胞，并发现肿瘤浸润性CD 8 + T细胞具有显著的 与血液CD 8 + T相比，与脂质过氧化和铁凋亡相关的基因表达增加 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 CD 8 + T细胞亚群、效应记忆（TEM）和终末分化效应（TTE）CD 8 + T细胞是 对肿瘤引起的铁凋亡更敏感。我们检测了来自骨骼的肿瘤浸润性CD 8 + T细胞， 多发性骨髓瘤患者的骨髓（BM;肿瘤床）。通过将T细胞分为幼稚、TTE或TEM 基于它们的CCR 7和CD 45 RA表达，我们发现TEM和TTE CD 8 + T细胞表达 更高水平的脂质过氧化和铁蛋白沉积相关基因，对肿瘤诱导的 与幼稚的CD 8 + T细胞相比，尽管它们表达类似水平的CD 36，但它们的铁凋亡率仍然很高。同样在 在小鼠黑色素瘤和MM模型中，增加的铁凋亡主要发生在肿瘤浸润性CD 8 + TEM中， TTE细胞，但与小肿瘤小鼠相比，大肿瘤小鼠的幼稚CD 8 + T细胞中没有 负担我们的离体研究证实，CD 8 + TEM和TTE细胞对肿瘤或FA-1细胞更敏感。 诱导的铁凋亡比幼稚T细胞和他们的细胞毒性细胞因子，如IFNγ和TNFα的生产， 压抑为了阐明潜在的机制，使用RNA-seq，并显示CD 8 + TEM和TTE 细胞表达的2，4-二烯酰辅酶A还原酶1（DECR 1）水平显著较低，DECR 1是一种限速酶， 多不饱和脂肪酸（PUFA）β-氧化，与幼稚CD 8 + T细胞相比。DECR 1的敲低（KD）， 幼稚的CD 8 + T细胞导致PUFA表达增加和过氧化物酶体功能障碍， 它们对肿瘤或FA诱导的铁凋亡的影响比对照T细胞要大。基于这些新发现，我们假设 CD 8 + TEM和TTE细胞，由于摄取更多的FA和减少DECR 1和PUFA的表达， 氧化，对肿瘤/TME诱导的铁凋亡敏感，抑制TEM和TTE细胞铁凋亡可能 有效提高免疫疗法对癌症患者的治疗效果。目标1将决定 TME中CD 8 + TEM和TTE细胞中肿瘤或FA诱导的铁凋亡的潜在机制。目标2将阐明 Fas在肿瘤和TME中蓄积及诱导CD 8+细胞脂质过氧化作用及其机制 TEM和TTE T细胞和Aim 3将抑制CD 8 + TEM和TTE细胞的铁凋亡，以增强抗癌疗效 免疫疗法实现这些目标将使我们深入了解这些机制， 肿瘤和TME如何诱导T细胞脂质过氧化以及铁凋亡如何介导T细胞凋亡 代谢紊乱和死亡了解这些机制非常重要， 帮助我们开发新的治疗方法，以靶向T细胞脂质代谢和TME， 提高癌症免疫治疗的疗效。