The zinc finger protein ZNF638 is a novel transcriptional regulator of thermogenesis - Resubmission

项目摘要/摘要 肥胖是由于储存和消耗的能量不平衡而引起的，并增加了患肥胖症的风险 新陈代谢紊乱，给我们的社会带来了巨大的经济负担。三种类型的脂肪细胞--白色， 棕色和米色--对维持卡路里储备和热量之间的平衡至关重要 被利用了。通过促进棕色和米色脂肪的热生成来增强能量消耗已被提出如下 对抗肥胖的策略；然而，目前已知的赋予生热能力的蛋白质武器库 对脂肪细胞的作用仍然有限。我们实验室发现锌指因子ZNF638是一种新的 脂肪细胞功能的转录调节因子。我们实验室进行的新工作现在提供了 有证据表明，脂肪组织中缺乏ZNF638的小鼠耐寒能力减弱 温度和肥胖倾向的增加。在这里，我们建议评估ZNF638的生理作用 并评估ZNF638功能丧失或获得对代谢的影响 功能障碍。此外，我们还计划确定调控ZNF638‘S水平的分子机制和 ZNF638通过它来协调调节能量平衡的转录途径。在… 完成后，这些研究将使ZNF638的S在FAT中的功能以及对ZNF638的特征的研究成为可能 其上游监管机构、下游目标启动子和转录伙伴关系，从而澄清了其 作用机制。为了执行拟议的工作，我们将利用新生成的ZNF638 FLOXED 以及我们实验室多年来使用的分子和细胞生物技术和试剂。 这些将与不偏不倚的方法相结合，如以下目标所述。在目标1中，我们将评估 ZNF638在能量消耗、血糖稳态和代谢控制中的生理作用 通过脂肪特异性ZNF638基因敲除和过度表达的小鼠和 将确定ZNF638对米色和棕色脂肪功能的遗传需求和充分性；在目标2 我们将确定在脂肪组织中调节ZNF638的转录和信号通路，并揭示 利用最先进的下一代测序技术，在脂肪中实现这种辅因子的全基因组靶标。在……里面 此外，我们还将确定ZNF638通过以下途径调节脂肪细胞转录的分子机制 能够使ZNF638‘S调控基因表达的新的关键转录伙伴的特征。 我们预计，概述的研究将为调节能量平衡的机制带来新的曙光，并将 最终允许定义新的策略来调节能量代谢，并可能对 抗肥胖疗法的发展。

PROJECT SUMMARY/ABSTRACT Obesity arises from an imbalance between the amount of energy stored and consumed and increases the risk of metabolic derangements, imposing significant economic burden to our society. Three types of fat cells -white, brown and beige- are critical for the maintenance of the equilibrium between calories hoarded and those utilized. Augmenting energy dissipation by boosting brown and beige fat thermogenesis has been proposed as a strategy to combat obesity; however, the current arsenal of proteins known to confer thermogenic competency to fat cells remains limited. Our laboratory has discovered that the zinc finger factor ZNF638 is a novel transcriptional regulator of adipocyte function. New work performed in our laboratory has now provided evidence demonstrating that mice lacking ZNF638 in fat tissue have weakened ability to withstand cold temperatures and increased propensity to obesity. Here we propose to assess the physiological role of ZNF638 in adipose tissues and to evaluate the consequences of loss- or gain-of-ZNF638-function on metabolic dysfunction. Furthermore, we plan to determine the molecular mechanisms that regulate ZNF638’s levels and functionality and through which ZNF638 coordinates transcriptional pathways regulating energy balance. At completion, the studies proposed will permit the detailing of ZNF638’s function in fat and the characterization of its upstream regulators, downstream target promoters and transcriptional partnerships, thereby clarifying its mechanism of action. To perform the work proposed we will take advantage of newly generated ZNF638 floxed mice and of molecular and cell biological techniques and reagents utilized in our laboratory over the years. These will be combined with unbiased approaches, as outlined in the aims below. In Aim 1 we will assess the physiological role of ZNF638 in the control of energy expenditure, glucose homeostasis and metabolic dysfunction through the characterization of adipose specific ZNF638 knock-out and overexpressing mice and will establish the genetic requirement and sufficiency of ZNF638 for beige and brown fat functionality; in Aim 2 we will identify the transcriptional and signaling pathways that regulate ZNF638 in adipose tissues and reveal the genome-wide targets of this cofactor in fat using state of the art next generation sequencing technologies. In addition we will define the molecular mechanisms through which ZNF638 regulates transcription in fat cells via the characterization of novel key transcriptional partners that enable ZNF638’s regulation of gene expression. We expect that the studies outlined will shed novel light into the mechanisms regulating energy balance and will ultimately permit the definition of new strategies to modulate energy metabolism with possible impact on the development of anti-obesity therapies.