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: 10313308
• 负责人: Graham Lobel
• 金额: $ 5.1万
• 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/10313308
• 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; Incubated; Inflammatory; Intercellular Fluid; Isotope Labeling; Knock-out; Knockout Mice; Label; Leucine; 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-Lymphocyte; Testing; Tetanus Helper Peptide; Time; Tumor Antigens; Tumor-associated macrophages; Tumor-infiltrating immune cells; Work; amino acid metabolism; anti-tumor immune response; antiport; base; 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/antagonist; insight; knock-down; liquid chromatography mass spectrometry; macrophage; monocyte; novel therapeutic intervention; protein activation; 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.

项目摘要 软组织肉瘤（STS）是一种多样的，往往是致命的恶性肿瘤， 转移性疾病的5年生存率为16%，反映了对新的治疗方法的需求。 战略布局一种有望对抗多种癌症的方法是免疫检查点阻断。然而，在这方面， 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 这些疾病的微环境特征。STS微环境由肿瘤主导- 相关的巨噬细胞，其在很大程度上与肿瘤浸润单核细胞分化。这些也可以 在炎症条件下分化为抗肿瘤单核细胞衍生的树突状细胞（Mo-DC），但Mo-DC STS中的分化受到未知因素的抑制。我们最近发现，增强型Mo-DC 分化导致与免疫检查点阻断的协同作用。因此，关键是要发现更多的 调节Mo-DC分化的过程，以提高针对STS的免疫疗法的功效。 一种这样的机制可能取决于谷氨酰胺代谢。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 脱氢酶、蛋白质糖基化和雷帕霉素复合物1（mTORC 1）的哺乳动物靶标的激活 介导的信号通路。此外，最近的研究表明，阻断谷氨酰胺代谢， 调节肿瘤浸润免疫细胞的抗肿瘤活性。使用Mo-DC的体外模型 我们发现，谷氨酰胺剥夺阻断了Mo-DC的分化，令人惊讶的是， 转氨酶活性。这一发现表明Mo-DC的分化需要谷氨酰胺通过单独的 途径，如己糖胺生物合成途径（HBP）或谷氨酰胺-亮氨酸反向转运，导致 mTORC 1激活。我推测谷氨酰胺代谢调节单核细胞衍生的树突状细胞 在软组织肉瘤微环境中通过己糖胺或 生物合成途径或亮氨酸依赖性mTORC 1激活。 目的1将确定谷氨酰胺代谢调节分化的机制， 单核细胞转化为Mo-DC。同位素标记和质谱法将用于评估 谷氨酰胺转化为谷氨酰胺酶的下游代谢物以及HBP中间体并从细胞输出。 条件性敲除小鼠将用于评估HBP通量和mTORC 1活化对Mo-12表达的贡献。 DC分化。目的2将测试谷氨酰胺可用性和谷氨酰胺代谢抑制对 Mo-DC在STS微环境中的分化和功能，以及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微环境中运行。