Novel transcriptional regulators for thermogenic program

产热程序的新型转录调节因子

• 批准号: 10318623
• 负责人: Hei Sook Sul
• 金额: $ 37.82万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-18 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318623
• 关键词: 3T3-L1 Cells; ATAC-seq; Ablation; Address; Adipose tissue; Adrenergic Agents; Adult; Affinity Chromatography; Binding; Biological; Biological Assay; Biology; Body Temperature; Brown Fat; CRISPR/Cas technology; Cells; ChIP-seq; Chromatin; Chronic Disease; Cre driver; Cyclic AMP-Dependent Protein Kinases; Diabetes Mellitus; Disease Management; EMSA; Energy Metabolism; Epidemic; Epigenetic Process; Family; Future; Gene Expression; Generations; Genes; Genetic Transcription; Glucose; Goals; Health; Homeostasis; Insulin; Insulin Resistance; Knockout Mice; Libraries; MAP Kinase Gene; Metabolic syndrome; Methods; Methyltransferase; Molecular; Morphology; Mus; Mutate; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Organ; Pathway interactions; Pattern; Phosphorylation; Physiological; Physiology; Prevention; Promoter Regions; Proteins; RNA Binding; RNA Recognition Motif; Regulation; Reporter; Reporting; Research; Resistance; Respiration; Role; System; Testing; Therapeutic; Thermogenesis; Tissues; Transcriptional Activation; Transgenic Mice; Zinc Fingers; bZIP Domain; cofactor; combat; diabetes control; diet-induced obesity; epigenetic regulation; gain of function; genome-wide; histone methyltransferase; improved; in vivo; inhibitor; interest; loss of function; methylation pattern; mouse model; mutant; new therapeutic target; novel; obesity treatment; overexpression; p38 Mitogen Activated Protein Kinase; pri-miRNA; programs; promoter; screening; transcription factor; transcriptome sequencing

Obesity, the accumulation of excess white adipose tissue (WAT), is associated with type 2 diabetes and other chronic diseases and has become an epidemic. While WAT is primarily an energy storage organ, brown adipose tissue (BAT) dissipates energy for heat generation via UCP1 to maintain body temperature. With the recognition of BAT/BAT-like tissues in human adults, increasing BAT or BAT activity may combat obesity. By screening a library of known and putative transcription factors that can activate UCP1 promoter, we recently have identified a largely uncharacterized a protein belonging to C3H zinc finger family, that can robustly activate UCP1 and other thermogenic genes. This transcription factor is enriched in BAT versus WAT and is induced upon cold-exposure. We detected binding of this C3H factor at the promoter regions of UCP1 and other BAT genes by EMSA and SELEX as well as by ChIP. Ablation by CRISPR-Cas9 system in BAT cells in culture suppressed UCP1 and other target genes to reduce uncoupled respiration. Moreover, overexpression in adipose tissue in mice increased energy expenditure to accompany resistance to diet- induced obesity, whereas ablation in adipose tissue promoted obesity and insulin resistance. To further examine the function of this factor, by TAP-Mass spec analysis, we identified a histone methyltransferase, which we found to interact and function synergistically. Adipose specific ablation of this epigenetic factor in mice also decreased energy expenditure promoting obesity. Our goal is to understand the molecular mechanisms and physiological significance of this new C3H transcription factor and its interacting protein for the thermogenic gene program and thermogenesis: Aim 1 is to investigate the function of this C3H factor in activating UCP1 promoter and study its genome-wide targets. Aim 2 is to examine the relationship and function of its interacting cofactor in epigenetic regulation of the thermogenic gene program. Finally, Aim 3 is to study the in vivo role of this C3H factor, its cofactor and their relationship in thermogenesis and adiposity by gain- and loss-of function studies in mice. Our research will help better understand the thermogenic program and may provide targets for developing effective obesity therapeutics in the future.

肥胖，即过量白色脂肪组织 (WAT) 的积累，与 2 型糖尿病和其他慢性疾病有关，并且已成为一种流行病。 WAT 主要是一个能量储存器官，而棕色脂肪组织 (BAT) 通过 UCP1 耗散能量产生热量以维持体温。随着对成人中 BAT/BAT 样组织的认识，增加 BAT 或 BAT 活性可能会对抗肥胖。通过筛选可以激活 UCP1 启动子的已知和推定转录因子文库，我们最近鉴定了一种很大程度上未表征的属于 C3H 锌指家族的蛋白质，它可以强力激活 UCP1 和其他产热基因。与 WAT 相比，该转录因子在 BAT 中富集，并且在冷暴露时被诱导。我们通过 EMSA 和 SELEX 以及 ChIP 检测到该 C3H 因子在 UCP1 和其他 BAT 基因的启动子区域的结合。培养中的 BAT 细胞中 CRISPR-Cas9 系统的消融抑制了 UCP1 和其他靶基因，从而减少了非偶联呼吸。此外，小鼠脂肪组织中的过度表达增加了能量消耗，伴随着对饮食诱导的肥胖的抵抗，而脂肪组织的消融则促进了肥胖和胰岛素抵抗。为了进一步检查该因子的功能，通过 TAP 质谱分析，我们鉴定了一种组蛋白甲基转移酶，我们发现该酶相互作用并具有协同作用。小鼠体内这种表观遗传因子的脂肪特异性消除也减少了促进肥胖的能量消耗。我们的目标是了解这种新的 C3H 转录因子及其相互作用蛋白对生热基因程序和生热作用的分子机制和生理意义：目标 1 是研究该 C3H 因子在激活 UCP1 启动子中的功能并研究其全基因组靶标。目标 2 是检查其相互作用的辅助因子在生热基因程序的表观遗传调控中的关系和功能。最后，目标 3 是通过小鼠功能获得和丧失的研究来研究这种 C3H 因子及其辅助因子的体内作用及其在产热和肥胖中的关系。我们的研究将有助于更好地了解生热程序，并可能为未来开发有效的肥胖疗法提供目标。

项目成果

Dot1l interacts with Zc3h10 to activate Ucp1 and other thermogenic genes.

• DOI: 10.7554/elife.59990
• 发表时间: 2020-10-27
• 期刊: eLife
• 影响因子: 7.7
• 作者: Yi D;Nguyen HP;Dinh J;Viscarra JA;Xie Y;Lin F;Zhu M;Dempersmier JM;Wang Y;Sul HS
• 通讯作者: Sul HS

Epigenetic dynamics of the thermogenic gene program of adipocytes.

• DOI: 10.1042/bcj20190599
• 发表时间: 2020-03-27
• 期刊: The Biochemical journal
• 作者: Yi D;Nguyen HP;Sul HS
• 通讯作者: Sul HS