SLC27A2, a critical metabolic regulator of T cellular inflammation in juvenile idiopathic arthritis and beyond?

SLC27A2，幼年特发性关节炎及其他疾病中 T 细胞炎症的关键代谢调节因子？

• 批准号: 498756914
• 负责人: Professor Dr. Klaus Tenbrock
• 金额: --
• 依托单位: Klinik für Kinder- und Jugendmedizin
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/498756914?language=en
• 关键词: SLC27A2; critical; metabolic; regulator; cellular

Juvenile idiopathic arthritis (JIA) is a disease of unknown origin in which cells of the innate immune system as well as of the adaptive immune system including T cells play central roles. T cellular function and fate decision critically depends on energetic properties including glycolysis or oxidative phosphorylation. Fatty acids are one of the most important sources of energy for oxidative phosphorylation. Within our preliminary data we show that1. the uptake of free fatty acids is enhanced in synovial fluid (SF) CD4+ T cells of patients with JIA, 2. the expression of SLC27A2/FATP2 is increased in SF CD4+ T cells, 3. the expression of SLC27A2 and uptake of free fatty acids is enhanced in blood derived CD4+ memory T cells after stimulation with anti-CD3/CD28, 4. the uptake of free fatty acids in anti-CD3/CD28-stimulated CD4+ T cells can be blocked with lipofermata, a specific inhibitor of SLC27A2 and5. the blockade with lipofermata inhibits T cell proliferation, IFNy production and mitochondrial respiration.Our data provide strong evidence that SLC27A2 might act as a critical fatty acid transport protein which is necessary for the proliferation and inflammatory propensities of memory T cells in JIA. This suggests that SLC27A2 is a novel therapeutic target in JIA and other autoimmune diseases. Several questions arise from the above-mentioned data:A)What are the functional consequences of fatty acid uptake in SF T cells and does this alter inflammatory cues within the cells? We will identify the fatty acid composition in SF using mass-spectrometry and perform metabolite analyzes after fatty acid uptake in JIA T cells. We will perform RNAseq of memory T cells and JIA SF T cells after incubation with lipofermata and stimulation with anti-CD3/CD28. B)Is T cell differentiation and function affected by SLC27A2 expression? Human and murine naïve CD4 T cells will be differentiated into Th1/Th2/Th17/Treg cells in the presence/absence of lipofermata as well as siRNA for SLC27A2. We will generate CD4CRESLC27A2fl/fl mice and analyze them with regard to T cell compartments, metabolism, transcriptional and function.C)What is the reason for SLC27A2 upregulation in JIA memory T cells? ATACseq will be performed in SF T cells as well as in stimulated and unstimulated memory T cells to decipher chromatin accessibility and to identify transcription factor binding sites within the promoter/enhancer regions.D)Can blockade of SLC27A2 prevent experimental arthritis? We will perform a Collagen- induced arthritis model in CD4CRESLC27A2fl/fl mice and controls as well as a T cell-mediated transfer arthritis model using lipofermata as therapeutic intervention. Our project will clarify the role of SLC27A2 for the energetic demands of CD4+ memory T cells in JIA. If our preliminary data hold true, we would have identified a novel concept of fatty acid uptake and metabolism in CD4+ memory T cells with the chance to specifically target it in inflammatory diseases.

幼年特发性关节炎 (JIA) 是一种来源不明的疾病，其中先天免疫系统以及包括 T 细胞在内的适应性免疫系统的细胞发挥着核心作用。 T 细胞功能和命运决定关键取决于能量特性，包括糖酵解或氧化磷酸化。脂肪酸是氧化磷酸化最重要的能量来源之一。在我们的初步数据中，我们表明1。 JIA 患者滑液 (SF) CD4+ T 细胞中游离脂肪酸的摄取增强，2. SF CD4+ T 细胞中 SLC27A2/FATP2 的表达增加，3. 用抗 CD3/CD28 刺激后，血源性 CD4+ 记忆 T 细胞中 SLC27A2 的表达和游离脂肪酸的摄取增强，4. 游离脂肪酸的摄取 抗 CD3/CD28 刺激的 CD4+ T 细胞中的酸可以被 lipofermata（一种 SLC27A2 和 5 的特异性抑制剂）阻断。 lipofermata 的阻断可抑制 T 细胞增殖、IFNy 产生和线粒体呼吸。我们的数据提供了强有力的证据，表明 SLC27A2 可能充当关键的脂肪酸转运蛋白，这对于 JIA 中记忆 T 细胞的增殖和炎症倾向是必需的。这表明 SLC27A2 是 JIA 和其他自身免疫性疾病的新治疗靶点。上述数据引发了几个问题：A) SF T 细胞摄取脂肪酸的功能后果是什么？这是否会改变细胞内的炎症信号？我们将使用质谱法鉴定 SF 中的脂肪酸组成，并在 JIA T 细胞摄取脂肪酸后进行代谢物分析。在与 lipofermata 孵育并用抗 CD3/CD28 刺激后，我们将对记忆 T 细胞和 JIA SF T 细胞进行 RNAseq。 B) T 细胞分化和功能是否受 SLC27A2 表达影响？在存在/不存在 lipofermata 以及 SLC27A2 的 siRNA 的情况下，人类和小鼠幼稚 CD4 T 细胞将分化为 Th1/Th2/Th17/Treg 细胞。我们将生成 CD4CRESLC27A2fl/fl 小鼠，并分析它们的 T 细胞区室、代谢、转录和功能。C) JIA 记忆 T 细胞中 SLC27A2 上调的原因是什么？ ATACseq 将在 SF T 细胞以及刺激和未刺激的记忆 T 细胞中进行，以破译染色质可及性并识别启动子/增强子区域内的转录因子结合位点。D) 阻断 SLC27A2 能否预防实验性关节炎？我们将在 CD4CRESLC27A2fl/fl 小鼠和对照小鼠中进行胶原诱导的关节炎模型，以及使用 lipofermata 作为治疗干预的 T 细胞介导的转移性关节炎模型。 我们的项目将阐明 SLC27A2 在满足 JIA 中 CD4+ 记忆 T 细胞能量需求方面的作用。如果我们的初步数据成立，我们将确定 CD4+ 记忆 T 细胞中脂肪酸摄取和代谢的新概念，并有机会专门针对炎症性疾病。