TRANSCRIPTIONAL REGULATION OF ADIPOCYTE INFLAMMATION BY EARLY B-CELL FACTOR-1 (Ebf1)

早期 B 细胞因子 1 (Ebf1) 对脂肪细胞炎症的转录调控

• 批准号: 9812728
• 负责人: Michael J Griffin
• 金额: $ 40.87万
• 依托单位: SAM HOUSTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-10 至 2023-09-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9812728
• 关键词: 3T3-L1 Cells; Abate; Adipocytes; Adipose tissue; Affect; Animal Model; Arizona; Arthritis; Atherosclerosis; B-Lymphocytes; Binding; Biological; Biological Assay; Biological Models; Biomedical Research; Boston; Cell Fractionation; Cells; Chronic; Chronic Disease; DNA Binding; DNA Binding Domain; Data; Data Analyses; Development; Diabetes Mellitus; Disease; Enterobacteria phage P1 Cre recombinase; Environment; Event; Fatty acid glycerol esters; Funding; Future; Gene Expression; Genes; Genetic Transcription; Genomics; Heart; Heart Diseases; High Fat Diet; Hyperplasia; Hypertension; Hypertrophy; Impairment; In Vitro; Individual; Inflammation; Inflammatory; Innate Immune System; Institution; Insulin Resistance; Israel; Knock-out; Knockout Mice; LoxP-flanked allele; Luciferases; Malignant Neoplasms; Medical Students; Medical center; Metabolic; Metabolic Diseases; Metabolic dysfunction; Modeling; Molecular; Molecular Biology; Molecular Biology Techniques; Mus; Obesity; Osteopathic Medicine; Overnutrition; Pathway interactions; Physiological; Proteins; RANTES; RNA; Receptor Signaling; Regulation; Reporter; Research; Reverse Transcription; Role; Science; Signal Pathway; Signal Transduction; Students; TYRP1 gene; Techniques; Time; Toll-Like Receptor Pathway; Toll-like receptors; Transactivation; Transcriptional Regulation; Universities; Veterinary Medicine; Work; adipocyte biology; adipokines; career; chemokine; chromatin immunoprecipitation; clinically relevant; college; cytokine; graduate student; in vivo; insight; insulin signaling; interest; knock-down; knockout animal; lentiviral-mediated; loss of function; new therapeutic target; novel therapeutics; p65; programs; promoter; protein B; protein protein interaction; protein purification; side effect; small hairpin RNA; summer research; synergism; transcription factor; university student

PROJECT SUMMARY/ABSTRACT Obesity, which is associated with hypertrophy and hyperplasia of adipocytes, is associated with many chronic diseases including insulin resistance and diabetes, heart disease, hypertension, and cancer. Chronic inflammation and activation of the innate immune system may lie at the heart of many of these diseases; overnutrition is known to cause inflammation in adipocytes and adipose tissue. However, the molecular mechanisms that initiate these inflammatory events in adipocytes are largely unknown, particularly at the transcriptional level. Previous work has shown that the transcription factor Early B-Cell Factor-1 (Ebf1) regulates numerous signaling pathways and promotes inflammation in adipocytes; however, the precise molecular mechanisms through which Ebf1 regulates expression of inflammatory loci remains unknown. We have recently discovered that Ebf1 can physically interact with multiple subunits of the well-known inflammatory transcription factor NF-κB, and many of the same inflammatory loci that are decreased in expression in Ebf1- deficient 3T3-L1 adipocytes are also known NF-κB target genes, suggesting that Ebf1 and NF-κB may work together to induce expression of inflammatory chemokines and other proteins in adipocytes. To determine the role of Ebf1 in vivo, and to investigate Ebf1 regulation of inflammatory pathways independent of its role in differentiation, we have developed Fat-Specific Ebf1 Knockout (FEBKO) mice. This model provides us a unique opportunity to examine the functional consequences of Ebf1 deficiency in the context of adipose tissue. The specific aims are to 1) fully characterize the Ebf1-NF-κB interaction, including mapping the interaction domains, determining if the interactions are direct, and whether the two proteins can synergistically activate NF-B reporter models; 2) determine if NF-B DNA binding and/or transactivation are impaired in Ebf1-deficient adipocytes; and 3) perform metabolic characterization of the FEBKO animals. These studies will provide insight into the molecular mechanisms governing expression of inflammatory chemokines and other proteins in adipocytes, and may uncover novel “druggable” targets in obesity and diabetes. Development of compounds that specifically disrupt the Ebf1-NF-B protein-protein interactions could potentially abate inflammation in adipocytes, with minimal side effects, since Ebf1 expression is relatively adipose-specific. Finally, the proposed research will provide abundant opportunities for Midwestern University (MWU) students to become involved in obesity/diabetes-related research, from in vitro bench work to in vivo physiological work to analysis and interpretation of data. This includes students from the Master’s in Biomedical Sciences (MBS) program, the Arizona College of Osteopathic Medicine (AZCOM) program, and possibly others (e.g., the new Veterinary Medicine program). This project will fill a much-needed niche for MWU students who are interested in participating in obesity/diabetes research, hopefully incite them to incorporate research into their careers, and will serve to strengthen the research environment at our institution at multiple levels.

项目总结/摘要 肥胖与脂肪细胞的肥大和增生有关， 慢性疾病，包括胰岛素抵抗和糖尿病、心脏病、高血压和癌症。慢性 炎症和先天免疫系统的激活可能是许多这些疾病的核心; 已知营养过剩会引起脂肪细胞和脂肪组织的炎症。然而，分子 在脂肪细胞中引发这些炎症事件的机制在很大程度上是未知的，特别是在 转录水平。先前的研究表明，转录因子早期B细胞因子-1（Ebf 1）调节 许多信号通路，并促进脂肪细胞的炎症;然而， Ebf 1调节炎症位点表达的机制仍不清楚。我们最近 发现Ebf 1可以与众所周知的炎症因子的多个亚基发生物理相互作用。 转录因子NF-κB，以及许多在Ebf 1中表达减少的相同炎症位点。 3 T3-L1缺陷型脂肪细胞也是已知的NF-κB靶基因，提示Ebf 1和NF-κB可能起作用 共同诱导脂肪细胞中炎性趋化因子和其它蛋白质的表达。 确定Ebf 1在体内的作用，并研究Ebf 1对炎症通路的调节 与其在分化中的作用无关，我们已经开发了脂肪特异性Ebf 1敲除（FEBKO）小鼠。该模型 为我们提供了一个独特的机会，以检查Ebf 1缺陷的功能后果的背景下， 脂肪组织具体目标是：1）充分表征Ebf 1-NF-κB相互作用，包括绘制 相互作用结构域，确定相互作用是否直接，以及两种蛋白质是否可以协同作用， 激活NF-κ B B报告模型; 2）确定NF-κ B B DNA结合和/或反式激活是否受损， Ebf 1缺陷型脂肪细胞;和3）进行FEBKO动物的代谢表征。这些研究将 提供了对炎症趋化因子和其他趋化因子表达的分子机制的深入了解， 这一发现可能揭示肥胖症和糖尿病中新的“可药物化”靶点。发展 特异性破坏Ebf 1-NF-κ B B蛋白-蛋白相互作用的化合物可能会减弱 在脂肪细胞中的炎症，具有最小的副作用，因为Ebf 1表达是相对脂肪特异性的。 最后，本研究将为中西部大学提供丰富的研究机会 学生参与肥胖/糖尿病相关的研究，从体外实验室工作到体内生理学 分析和解释数据的工作。这包括来自生物医学科学硕士（MBS）的学生 计划，亚利桑那大学骨病医学（AZCOM）计划，以及可能的其他（例如，新 兽医专业）。这个项目将填补一个急需的利基为MWU学生谁有兴趣 参与肥胖/糖尿病研究，希望能激励他们将研究纳入自己的职业生涯， 将有助于在多个层面上加强我们机构的研究环境。