Targeting T cell glutamine metabolism in severe asthma

针对严重哮喘患者的 T 细胞谷氨酰胺代谢

• 批准号: 10211394
• 负责人: Dawn C Newcomb
• 金额: $ 60.31万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-06 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10211394
• 关键词: Affect; Airway Disease; Allergens; Amino Acids; Animal Model; Asthma; Automobile Driving; Bronchial Spasm; CD4 Positive T Lymphocytes; CRISPR screen; Cell physiology; Cells; Cellular Metabolic Process; Characteristics; Chromatin; Clinic; Clinical; Cytometry; Data; Dependence; Development; Disease; Electron Transport; Enzymes; Epigenetic Process; Equilibrium; Estrogen Receptors; Female; Frequencies; Gene Expression; Gene Targeting; Generations; Genes; Genetic; Glucose; Glutamates; Glutaminase; Glutamine; Glycolysis; Guanine Nucleotide Exchange Factors; Hexosamines; High Fat Diet; IL17 gene; IL4 gene; Immune; In Vitro; Incidence; Inflammation; Inflammatory; Interleukin-17; Liquid substance; Lung; Lung Inflammation; Lung Lavage Fluid; Mediating; Metabolic; Metabolic Diseases; Metabolic Marker; Metabolic Pathway; Metabolic dysfunction; Metabolism; Mitochondria; Modeling; Mucous body substance; Mus; Obesity; Oxidation-Reduction; Pathway interactions; Patients; Play; Population Process; Production; Pyroglyphidae; Regulatory T-Lymphocyte; Resistance; Respiration; Risk; Risk Factors; Role; Sampling; Severities; T cell differentiation; T cell regulation; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Th2 Cells; Therapeutic; Translating; Weight; Woman; airway hyperresponsiveness; airway inflammation; airway remodeling; allergic airway inflammation; alpha ketoglutarate; asthma exacerbation; asthmatic patient; chronic inflammatory disease; cytokine; diet-induced obesity; effector T cell; eosinophil; glucose metabolism; health disparity; high dimensionality; immune function; in vivo; inhibitor/antagonist; male; men; metabolomics; mitochondrial metabolism; mouse model; neutrophil; new therapeutic target; novel therapeutics; obesity risk; peripheral blood; programs; sex; therapy resistant; uptake

SUMMARY Asthma is a debilitating disease of airway inflammation with increasing worldwide occurrence. With higher incidence in females, asthma is exacerbated by obesity and metabolic disease. Underlying asthma is activation of innate immune and T cells that drive inflammation. Airway inflammation in severe asthma is caused in part by an imbalance between effector CD4 T cells (including Th2 and Th17) and suppressive regulatory (Treg) T cells. This imbalance is particularly heightened in obese females and leads to increased eosinophils and/or neutrophils in the airway, increased airway hyperresponsiveness, and increased mucus production. It is now clear that T cell metabolism plays a critical role to regulate immune function and the distinct metabolic programs of each T cell subset can influence inflammation and play roles in asthma. In addition to a requirement of T cells for glycolysis, our data show that glutamine metabolism plays a key role to support inflammatory T cells in asthma. Metabolomics data from bronchoalveolar fluid and mass cytometry of T cells from donors with severe asthma showed markers of elevated glutaminolysis. While IL17 producing cells both in vitro and from lungs had the highest expression of metabolic markers, both Th17 and Th2 cells relied on glutamine for differentiation and cytokine production. Glutamine metabolism is regulated through uptake by the transporter ASCT2 and Glutaminase (GLS)-dependent conversion to glutamate and α-ketoglutarate that supports mitochondrial electron transport and regulates epigenetic marks that control chromatin and gene expression. We have now shown through inhibition, gene targeting, and in vivo primary T cell CRISPR screening in lung inflammation that ASCT2 and GLS are essential for Th17 and Th2 cells. These genes are dispensable for Treg and blocking glutamine uptake can instead enhance Treg differentiation. Importantly, we also show in an animal model that high fat diet fed female mice have selectively increased frequencies of Th17 cells. This effect is sex-specific, as high fat diet fed female, but not male, mice developed increased IL17 production and we show that the Estrogen Receptor (ERα) plays a key role in females to elevate CD4 T cell glycolysis and mitochondrial metabolism. We hypothesize that airway allergen exposure and inflammatory cytokines induce T cell glutamine metabolism, driving Th2 and Th17 cells that contribute to asthma incidence and severity in obese females. To test this hypothesis, we will: (1) Determine the role and mechanism of GLS and ASCT2-dependent glutamine metabolism to regulate the balance of Th2, Th17 and Treg cells in a mixed model of allergic airway inflammation and (2) Test if obesity- induced exacerbation of asthma severity in females is dependent on enhanced T cell glutamine metabolism in animal models and from peripheral blood and excised lungs of healthy and severe asthma normal weight or obese donors. These studies will establish metabolic mechanisms and if inhibiting glutamine uptake or metabolism through ASCT2 or GLS can shift the balance of Th17 and Th2 cells to instead favor Treg and protect against asthma to help alleviate this health disparity observed in females with metabolic disease.

摘要 哮喘是一种使人衰弱的呼吸道炎症疾病，在全球范围内发病率越来越高。具有更高的 在女性中，肥胖和代谢性疾病会加重哮喘的发病率。哮喘的基础是激活 先天免疫和驱动炎症的T细胞。重症哮喘患者的呼吸道炎症部分是由 效应性CD4T细胞(包括Th2和Th17)和抑制性调节性T细胞(Treg)之间的失衡。 这种失衡在肥胖女性中尤其严重，并导致嗜酸性粒细胞和/或中性粒细胞增加。 在呼吸道中，呼吸道高反应性增加，粘液产生增加。现在很清楚，T细胞 新陈代谢对调节免疫功能和每个T细胞不同的新陈代谢程序起着关键作用 亚群可影响炎症反应，在哮喘发病中发挥作用。除了糖酵解需要T细胞外， 我们的数据表明，谷氨酰胺代谢在支持哮喘中的炎性T细胞方面起着关键作用。 支气管肺泡液代谢组学数据和重症哮喘供者T细胞质量细胞学分析 显示出谷氨酰胺分解升高的标志。而在体外和从肺中产生IL17的细胞具有 代谢标志物的最高表达，Th17和Th2细胞均依赖谷氨酰胺分化和 细胞因子的产生。谷氨酰胺代谢通过转运体ASCT2和 谷氨酰胺酶依赖的谷氨酸和支持线粒体电子的α-酮戊二酸的转化 运输和调节控制染色质和基因表达的表观遗传标记。我们现在已经展示了 通过抑制、基因打靶和体内原代T细胞CRISPR筛选ASCT2在肺部炎症中的作用 和GLS是Th17和Th2细胞所必需的。这些基因对于Treg和阻断谷氨酰胺来说是不必要的 相反，摄取可以促进Treg的分化。重要的是，我们还在动物模型中表明，高脂肪饮食 喂食雌性小鼠有选择地增加了Th17细胞的频率。这种影响是因性别而异的，因为高脂肪饮食 喂食雌性而不是雄性的小鼠产生更多的IL17，我们发现雌激素受体 ER-α在女性提高CD4T细胞糖酵解和线粒体代谢中起着关键作用。我们假设 呼吸道过敏原暴露和炎性细胞因子诱导T细胞谷氨酰胺代谢，驱动Th2和 Th17细胞与肥胖女性哮喘的发病率和严重程度有关。为了验证这一假设，我们将： (1)确定GLS和ASCT2依赖的谷氨酰胺代谢在调节血管紧张素转换酶中的作用和机制 过敏性呼吸道炎症混合模型中Th2、Th17和Treg细胞的平衡以及(2)肥胖- 女性哮喘严重程度的诱导加重依赖于T细胞谷氨酰胺代谢的增强 正常体重或正常体重的健康和重度哮喘动物模型及外周血和离体肺 肥胖的捐赠者。这些研究将建立代谢机制，如果抑制谷氨酰胺摄取或 通过ASCT2或GLS的代谢可以改变Th17和Th2细胞的平衡，而不是有利于Treg和保护 对抗哮喘，以帮助缓解在患有代谢性疾病的女性中观察到的这种健康差距。