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

项目总结/摘要 肥胖是由于能量储存和消耗之间的不平衡而引起的， 代谢紊乱，给我们的社会带来巨大的经济负担。三种脂肪细胞-白色， 棕色和米色--对于维持囤积的卡路里和那些 利用。已经提出通过促进棕色和米色脂肪产热来增加能量耗散， 一种对抗肥胖的策略;然而，目前已知赋予产热能力的蛋白质库 对脂肪细胞的影响仍然有限。我们实验室发现锌指因子ZNF 638是一种新的 脂肪细胞功能的转录调节因子。我们实验室的新工作提供了 有证据表明，脂肪组织中缺乏ZNF 638的小鼠耐寒能力减弱 温度升高和肥胖倾向增加。在这里，我们建议评估ZNF 638的生理作用， 并评估ZNF 638功能丧失或获得对代谢的影响。 功能障碍此外，我们计划确定调节ZNF 638水平的分子机制， ZNF 638通过其功能协调调节能量平衡的转录途径。在 完成后，拟议的研究将允许详细说明ZNF 638在脂肪中的功能，并表征 它的上游调节子，下游靶向启动子和转录伙伴关系，从而阐明了它的 作用机制。为了执行建议的工作，我们将利用新生成的ZNF 638 floxed 小鼠以及多年来我们实验室使用的分子和细胞生物学技术和试剂。 这些将与公正的方法相结合，如下面的目标所述。在目标1中，我们将评估 ZNF 638在控制能量消耗、葡萄糖稳态和代谢中的生理作用 通过对脂肪特异性ZNF 638敲除和过表达小鼠的表征， 将确定ZNF 638对米色和棕色脂肪功能的遗传要求和充足性;在目标2中， 我们将确定在脂肪组织中调控ZNF 638的转录和信号通路，并揭示 使用最先进的下一代测序技术，在脂肪中的这种辅因子的全基因组靶点。在 此外，我们还将通过以下途径确定ZNF 638调节脂肪细胞转录的分子机制： 新的关键转录伙伴，使ZNF 638的基因表达调控的表征。 我们希望，概述的研究将揭示新的光调节能量平衡的机制， 最终允许定义新的策略来调节能量代谢，并可能对人体产生影响 开发抗肥胖疗法。

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.