The Role of Glutamine Metabolism in Monocyte-Derived Dendritic Cell Differentiation in the Soft-Tissue Sarcoma Microenvironment

谷氨酰胺代谢在软组织肉瘤微环境中单核细胞衍生的树突状细胞分化中的作用

• 批准号: 10668447
• 负责人: Graham Lobel
• 金额: $ 3.51万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-08 至 2026-07-07
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10668447
• 关键词: Amino Acids; Antigen-Presenting Cells; Cell Differentiation process; Cell physiology; Cells; Cessation of life; Characteristics; Complex; Connective Tissue; Dendritic Cells; Dependence; Disease; Doxycycline; Engineering; Enzymes; FRAP1 gene; Flow Cytometry; Generations; Genetic Transcription; Glucosamine; Glutamates; Glutaminase; Glutamine; Granulocyte-Macrophage Colony-Stimulating Factor Receptors; Hexosamines; Immune; Immune signaling; Immune system; Immunologic Receptors; Immunotherapeutic agent; Immunotherapy; Implant; Incubated; Infiltration; Inflammatory; Intercellular Fluid; Isotope Labeling; Knock-out; Knockout Mice; Label; Leucine; Macrophage; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Metabolic; Metabolic Pathway; Metabolism; Modeling; Mus; Myelogenous; Myeloid Cells; Pathway interactions; Population; Process; Production; Protein Glycosylation; Resistance; Role; Shapes; Signal Pathway; Sirolimus; Soft tissue sarcoma; Survival Rate; T-Cell Activation; T-Lymphocyte; Testing; Tetanus Helper Peptide; Time; Tumor Antigens; Tumor-associated macrophages; Tumor-infiltrating immune cells; Work; amino acid metabolism; antagonist; anti-tumor immune response; antiport; biosynthetic product; cancer immunotherapy; cell type; conditional knockout; cytokine; deprivation; fibrosarcoma; glycosylation; immune checkpoint blockade; immune function; immunosuppressive macrophages; immunotherapy clinical trials; improved; in vitro Model; inhibitor; insight; knock-down; liquid chromatography mass spectrometry; monocyte; novel therapeutic intervention; receptor; synergism; transcriptome sequencing; tumor; tumor growth; tumor microenvironment; tumor-immune system interactions

PROJECT SUMMARY Soft-tissue sarcomas (STS) are a diverse and often fatal set of malignancies arising from connective tissue with a 16% five-year survival rate for metastatic disease, reflecting the need for novel therapeutic strategies. One approach showing promise against multiple cancers is immune checkpoint blockade. However, clinical trials of immunotherapy in STS have produced disappointing results, likely due to immunosuppressive microenvironments characteristic of these diseases. The STS microenvironment is dominated by tumor- associated macrophages, which largely differentiate from tumor-infiltrating monocytes. These can also differentiate into anti-tumor monocyte-derived dendritic cells (Mo-DCs) in inflammatory conditions, but Mo-DC differentiation is inhibited in STS by as-yet unknown factors. We have recently shown that enhanced Mo-DC differentiation leads to synergy with immune checkpoint blockade. It is therefore critical to discover additional processes that regulate Mo-DC differentiation in order to improve the efficacy of immunotherapy against STS. One such mechanism may depend on glutamine metabolism. Glutamine is utilized as a metabolic fuel by several immune cell types, as it is involved in generating biosynthetic products via the rate-limiting enzyme glutaminase, glycosylation of proteins, and activation of mammalian target of rapamycin complex 1 (mTORC1) mediated signaling pathways. Additionally, recent work has shown that blocking glutamine metabolism can modulate the anti-tumor activity of tumor-infiltrating immune cells. Using an in vitro model of Mo-DC differentiation, we found that glutamine deprivation blocked Mo-DC differentiation, surprisingly independent of glutaminase activity. This finding suggests Mo-DC differentiation requires glutamine flux through separate pathway(s), such as the hexosamine biosynthetic pathway (HBP) or glutamine-leucine antiport leading to mTORC1 activation. I hypothesize that glutamine metabolism regulates monocyte-derived dendritic cell differentiation in the soft tissue sarcoma microenvironment through either the hexosamine biosynthetic pathway or leucine-dependent mTORC1 activation. Aim 1 will identify the mechanisms by which glutamine metabolism regulates the differentiation of monocytes into Mo-DCs. Isotopic labeling and mass spectrometry will be used to assess the incorporation of glutamine into downstream metabolites of glutaminase as well as HBP intermediates and export from the cell. Conditional knockout mice will be used to assess the contributions of HBP flux and mTORC1 activation to Mo- DC differentiation. Aim 2 will test the impact of glutamine availability and inhibition of glutamine metabolism on Mo-DC differentiation and function in the STS microenvironment, as well as synergy with immune checkpoint blockade. Together, these approaches will elucidate the role of glutamine metabolism in Mo-DC differentiation and function in the STS microenvironment.

项目总结 软组织肉瘤(STS)是一种由结缔组织瘤引起的多种多样且常常致命的恶性肿瘤。 转移性疾病的五年存活率为16%的组织，反映了对新疗法的需求 战略。一种显示出对抗多种癌症的前景的方法是免疫检查点封锁。然而， STS免疫治疗的临床试验结果令人失望，可能是由于免疫抑制 这些疾病特有的微环境。STS微环境由肿瘤主导- 相关巨噬细胞，与肿瘤浸润性单核细胞有很大区别。这些也可以 在炎症条件下可分化为抗肿瘤单核细胞来源的树突状细胞(Mo-DC)，但Mo-DC 在STS中，分化受到未知因素的抑制。我们最近证明了增强的钼-直流电 分化导致与免疫检查点阻断的协同作用。因此，关键是要发现其他 调节Mo-DC分化以提高免疫治疗对STS的疗效的过程。 其中一种机制可能依赖于谷氨酰胺代谢。谷氨酰胺被用作代谢燃料 通过几种免疫细胞类型，因为它参与通过限速酶产生生物合成产物 谷氨酰胺酶、蛋白质糖基化和哺乳动物雷帕霉素复合体靶标1(MTORC1)的激活 介导的信号通路。此外，最近的研究表明，阻止谷氨酰胺代谢可以 调节肿瘤浸润性免疫细胞的抗肿瘤活性。利用钼-树突状细胞体外模型 分化，我们发现谷氨酰胺剥夺阻断了Mo-DC的分化，令人惊讶地独立于 谷氨酰胺酶活力。这一发现表明，Mo-DC的分化需要谷氨酰胺通过单独的 途径(S)，如氨基己糖生物合成途径(HBP)或谷氨酰胺-亮氨酸反向通道导致 MTORC1激活。我假设谷氨酰胺代谢调节单核细胞来源的树突状细胞 氨基己糖对软组织肉瘤微环境的分化作用 生物合成途径或依赖亮氨酸的mTORC1激活。 目标1将确定谷氨酰胺代谢调节细胞分化的机制 单核细胞转化为钼树突状细胞。将使用同位素标记和质谱学来评估纳入 谷氨酰胺进入谷氨酰胺酶的下游代谢物以及HBP中间产物并从细胞中输出。 条件基因敲除小鼠将被用来评估HBP通量和mTORC1激活对Mo-1的贡献。 DC分化。目的2将测试谷氨酰胺的可用性和谷氨酰胺代谢的抑制对 Mo-DC在STS微环境中的分化和功能及其与免疫检查点的协同作用 封锁。总之，这些方法将阐明谷氨酰胺代谢在Mo-DC分化中的作用 并在STS微环境中发挥作用。