Department of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston, Texas, USA.
Department of Pathology and Genomic Medicine, Center for Molecular and Translational Human Infectious Diseases Research Houston Methodist Research Institute, Weill-Cornell Medicine, Houston, Texas, USA.
J Biol Chem. 2022 Oct;298(10):102418. doi: 10.1016/j.jbc.2022.102418. Epub 2022 Aug 27.
Macrophages (MФ) are an essential immune cell for defense and repair that travel to different tissues and adapt based on local stimuli. A critical factor that may govern their polarization is the crosstalk between metabolism and epigenetics. However, simultaneous measurements of metabolites, epigenetics, and proteins (phenotype) have been a major technical challenge. To address this, we have developed a novel triomics approach using mass spectrometry to comprehensively analyze metabolites, proteins, and histone modifications in a single sample. To demonstrate this technique, we investigated the metabolic-epigenetic-phenotype axis following polarization of human blood-derived monocytes into either 'proinflammatory M1-' or 'anti-inflammatory M2-' MФs. We report here a complex relationship between arginine, tryptophan, glucose, and the citric acid cycle metabolism, protein and histone post-translational modifications, and human macrophage polarization that was previously not described. Surprisingly, M1-MФs had globally reduced histone acetylation levels but high levels of acetylated amino acids. This suggests acetyl-CoA was diverted, in part, toward acetylated amino acids. Consistent with this, stable isotope tracing of glucose revealed reduced usage of acetyl-CoA for histone acetylation in M1-MФs. Furthermore, isotope tracing also revealed MФs uncoupled glycolysis from the tricarboxylic acid cycle, as evidenced by poor isotope enrichment of succinate. M2-MФs had high levels of kynurenine and serotonin, which are reported to have immune-suppressive effects. Kynurenine is upstream of de novo NAD metabolism that is a necessary cofactor for Sirtuin-type histone deacetylases. Taken together, we demonstrate a complex interplay between metabolism and epigenetics that may ultimately influence cell phenotype.
巨噬细胞（MФ）是一种重要的免疫细胞，用于防御和修复，可迁移到不同的组织，并根据局部刺激进行适应性改变。可能支配其极化的一个关键因素是代谢和表观遗传学之间的串扰。然而，代谢物、表观遗传学和蛋白质（表型）的同时测量一直是一个主要的技术挑战。为了解决这个问题，我们开发了一种新的三omics 方法，使用质谱法在单个样本中全面分析代谢物、蛋白质和组蛋白修饰。为了证明这一技术，我们研究了人血衍生单核细胞极化为“促炎 M1-”或“抗炎 M2-”MФ 后代谢-表观遗传-表型轴。我们在这里报告了精氨酸、色氨酸、葡萄糖和柠檬酸循环代谢、蛋白质和组蛋白翻译后修饰以及人类巨噬细胞极化之间以前未描述的复杂关系。令人惊讶的是，M1-MФ 具有全局降低的组蛋白乙酰化水平，但具有高水平的乙酰化氨基酸。这表明乙酰辅酶 A 被部分转移到乙酰化氨基酸中。与此一致，葡萄糖的稳定同位素示踪表明 M1-MФ 中用于组蛋白乙酰化的乙酰辅酶 A 减少。此外，同位素示踪还表明 MФ 使糖酵解与三羧酸循环解偶联，这表现为琥珀酸的同位素丰度增加不佳。M2-MФ 具有高水平的犬尿氨酸和血清素，据报道它们具有免疫抑制作用。犬尿氨酸是从头 NAD 代谢的上游物质，是 Sirtuin 型组蛋白去乙酰化酶的必要辅助因子。总之，我们展示了代谢和表观遗传学之间的复杂相互作用，这可能最终影响细胞表型。
