Epigenetics of Severe Systemic Inflammation

严重全身炎症的表观遗传学

• 批准号: 7847303
• 负责人: Charles Emory McCall
• 金额: $ 37万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-05 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7847303
• 关键词: Acute; Anti-Inflammatory Agents; Anti-inflammatory; Binding; Biochemical; Blood; Cell model; Characteristics; Chromatin; Clinical; Coupled; Cytosine; DNA; DNA Binding; DNA Modification Methylases; DNA Sequence; Data; Disease; Epigenetic Process; Euchromatin; Gene Expression; Gene Silencing; Genes; Genetic; Genetic Transcription; Heterochromatin; Histone Code; Histone H3; Histones; Human; Hypermethylation; Immunoblotting; Immunosuppression; Inflammation; Inflammatory; Interleukin-1; Leukocytes; Lysine; Mass Spectrum Analysis; Messenger RNA; Methylation; Methyltransferase; Nucleosomes; Outcome; Patients; Phosphotransferases; Plasmids; Process; Protein Dephosphorylation; Protein-Serine-Threonine Kinases; Proteins; Public Health; RPS6KA5 gene; Reporting; Repression; Sepsis; Serine; Small Interfering RNA; TNF gene; Testing; Tissues; Transfection; Transferase; Translational Research; Trauma; abstracting; adapter protein; antimicrobial peptide; base; chromatin immunoprecipitation; cytokine; gene repression; genetic regulatory protein; heterochromatin-specific nonhistone chromosomal protein HP-1; histone methyltransferase; human disease; improved; novel therapeutic intervention; promoter; public health relevance; response

Abstract Epigenetic gene reprogramming is an emerging concept relevant to human diseases. This proposal's objective is to investigate epigenetics of severe systemic inflammation (SSI). SSI blood and tissue leukocytes show gene-specific reprogramming with repressed transcription of acute proinflammatory genes like TNF and IL-1 and activated transcription of other gene sets. SSI gene reprogramming reversal correlates with clinical improvement. We reported TNF transcription repression and a nucleosome shift to lysine 9 H3 di-methylation (H3K9me2), H3 serine 10 dephosphorylation(H3S10), and repressor heterochromatin protein 1 (HP-1) promoter binding in an SSI cell model. We further implicate the histone H3 lysine methyltransferase G9a coupled to HP-1 and CpG methylation by cytosine methyltransferase DNMT 3a/b during TNF silencing. Together, these results support that epigenetic mechanisms participate in repressing acute inflammatory genes and sustaining often lethal immunosuppression observed in SSI patients. We hypothesize that SSI generates a shift from a euchromatin (responsive) heterochromatin (silenced) at the proximal promoter of acute proinflammatory genes. Aim 1 will test in whether the euchromatin to heterochromatin shift requires specific H3 serine kinases, lysine methyltransferases and demethylases, and adapters or linkers, and the potential for reversibility. Aim 2 will test whether acute proinflammatory gene promoter CpG methylation is coupled through specific regulatory proteins to the nucleosome shifts. Aim 3 will test whether the epigenetic shift from euchromatin to heterochromatin occurs in circulating human SSI leukocytes. Experimental approaches for the basic (Aims 1 and 2) and translational aims (Aim 3) will include genetic (transfection of siRNA or expression plasmids) and biochemical (chromatin immunoprecipitation, DNA sequencing, DNAase footprinting, immunoblots, and mRNA quantitation) analyses. *

摘要 表观遗传基因重编程是与人类疾病相关的一个新兴概念。这个提议 目的探讨严重全身性炎症（SSI）的表观遗传学。SSI血液和组织白细胞 显示基因特异性重编程，抑制急性促炎基因如TNF的转录， IL-1和其他基因组的激活转录。SSI基因重编程逆转与临床 改进.我们报道了肿瘤坏死因子转录抑制和核小体转移到赖氨酸9 H3二甲基化 （H3 K9 me 2）、H3丝氨酸10去磷酸化（H3 S10）和阻遏异染色质蛋白1（HP-1） SSI细胞模型中的启动子结合。我们进一步暗示组蛋白H3赖氨酸甲基转移酶G9 a 在TNF沉默期间，通过胞嘧啶甲基转移酶DNMT 3a/B偶联HP-1和CpG甲基化。 总之，这些结果支持表观遗传机制参与抑制急性炎症反应。 在SSI患者中观察到的基因和持续的通常致命的免疫抑制。我们假设SSI 产生从常染色质（响应性）异染色质（沉默）在近端启动子的转变， 急性促炎基因Aim 1将测试常染色质向异染色质的转移是否需要 特异性H3丝氨酸激酶、赖氨酸甲基转移酶和脱甲基酶，以及衔接子或接头，以及 可逆性的潜力。目的2将检测急性促炎基因启动子CpG甲基化是否与急性炎症相关。 通过特定的调节蛋白与核小体移位偶联。目标3将测试表观遗传是否 从常染色质到异染色质的转变发生在循环的人SSI白细胞中。实验 用于基本（目标1和2）和转化目标（目标3）的方法将包括遗传（转染 siRNA或表达质粒）和生物化学（染色质免疫沉淀、DNA测序、DNA酶 足迹法、免疫印迹和mRNA定量）分析。 *