Novel Thermogenic Factors to Combat Obesity and Metabolic Disease

项目摘要 肥胖是糖尿病和心血管疾病等慢性疾病发展的重要危险因素 疾病目前的治疗是无效的，因为肥胖率在世界范围内增加，现在流行 比例。世界卫生组织（WHO）报告称，全球有19亿人（三分之一） 超重和控制肥胖相关疾病的成本上升占年度支出的1470亿美元 美国2008年医疗保健支出。需要额外的治疗选择是至关重要的。这个目标 该提案旨在开发肥胖和相关代谢疾病的分子治疗方法。我们利用 产热棕色和米色脂肪生物学，当有针对性时，可以增加能量消耗， 通过解偶联蛋白1（UCP 1）的作用防止肥胖。这个过程的机制， 然而，还没有完全理解。揭示新的途径和调节因子，可以激活这一点， 该方法将使患有肥胖症和相关代谢疾病的个体的治疗选择最大化。 我们先前在小鼠模型中的工作表明转录因子Yin Yang 1（YY 1）是关键的 棕色脂肪的功能。脂肪特异性YY 1消融（YY 1bKO）导致显著的产热缺陷， 棕色脂肪组织，但机制尚不清楚。为了深入了解作用机制，我们 在YY 1bKO棕色脂肪中进行无偏转录组学分析，以寻找新的产热调节剂， 鉴定了基因nipsnap 1（4-硝基苯基磷酸酶结构域和非神经元SNAP-25样蛋白 同源物1）。Nipsnap 1在物种间进化上是保守的，但仍需要一个明确的功能注释。 分配。我们的初步数据表明，Nipsnap 1表现出强烈的产热特征，并显着改变 UCP 1蛋白水平。因此，本提案的目标是确定Nipsnap 1的分子功能， 产热脂肪组织，以检验Nipsnap 1在 产热作用这项研究将使用生物化学方法在原代棕色和米色脂肪细胞 详细描述Nipsnap 1在各种致热刺激下的转录和翻译动力学 （目标1）。然后，我们将继续通过获得和CRISPR丢失对Nipsnap 1进行功能表征， 功能研究除了测试偶联和非偶联的线粒体呼吸能力外， 系统还将使用高效LC-MS/MS评估产热脂肪代谢组学分析 (Aim 2）。我们的研究将是第一个表征分子功能以及代谢组学相关的 Nipsnap 1在产热脂肪组织中的网络。这项提案的结果将决定Nipsnap 1 可以用于治疗肥胖症和相关的代谢紊乱。

PROJECT SUMMARY Obesity is a significant risk factor for the development of chronic disorders such as diabetes and cardiovascular disease. Current treatments are ineffectual given the worldwide increase in obesity rates to now epidemic proportions. The World Health Organization (WHO) reports that 1.9 billion (1 in 3) individuals globally are overweight and the rising cost to manage obesity-related diseases represented $147 billion dollars of the annual U.S. healthcare expenditure in 2008. The need for additional treatment options is critical. The goal of this proposal is to develop molecular treatments for obesity and associated metabolic disease. We take advantage of the thermogenic brown and beige fat biology that, when targeted, can increase energy expenditure and confer protection against obesity through the action of uncoupling protein 1 (UCP1). The mechanisms of this process, however, are not completely understood. Unveiling novel pathways and regulatory factors that can activate this process will maximize treatment options for individuals suffering from obesity and associated metabolic disease. Our previous work in murine models demonstrated that the transcription factor Yin Yang 1 (YY1) is critical for brown fat function. Adipose-specific ablation of YY1 (YY1bKO) results in significant thermogenic defects in the brown fat tissue, but the mechanism is unknown. In order to gain insight into the mechanism of action, we performed unbiased transcriptomic profiling in YY1bKO brown fat in search for novel thermogenic regulators and identified the gene nipsnap1 (4-nitrophenyl phosphatase domain and non-neuronal SNAP-25 like protein homolog1). Nipsnap1 is evolutionarily conserved across species, yet a clear functional annotation remains to be assigned. Our preliminary data show that Nipsnap1 exhibits a strong thermogenic profile and dramatically alters UCP1 protein levels. The goal of this proposal therefore is to identify the molecular function of Nipsnap1 in thermogenic adipose tissue in order to test the hypothesis that Nipsnap1 plays a significant role in thermogenesis. The studies proposed will use biochemical approaches in primary brown and beige adipocytes to detail the transcription and translation dynamics of Nipsnap1 in response to various thermogenic stimuli (Aim1). We will then continue with the functional characterization of Nipsnap1 through gain- and CRISPR-loss- of-function studies in addition to testing the coupled and uncoupled mitochondrial respiratory capacity of the system. Thermogenic adipose metabolomic profiling using high performance LC-MS/MS will also be assessed (Aim 2). Our studies will be the first to characterize the molecular function as well as the metabolomic-linked network of Nipsnap1 in thermogenic adipose tissue. The findings from this proposal will determine if Nipsnap1 can be leveraged towards a treatment for obesity and associated metabolic disorders.