Development and validation of a human brown adipocyte system

 描述（由申请人提供）：棕色脂肪组织的激活和募集已成为对抗肥胖及其相关代谢疾病的令人兴奋的靶点。棕色脂肪组织与白色脂肪组织的不同之处在于其通过UCP-1驱动的适应性产热将能量作为热量燃烧的关键能力。线粒体UCP-1的表达使氧化磷酸化解偶联，导致底物氧化加速，但ATP产生减少。由此产生的能量耗散与血浆甘油三酯清除相关，这是体重减轻的关键因素。长期以来，人们就知道棕色脂肪库存在于啮齿动物和人类婴儿中。然而，最近的证据表明，在成人中存在活跃的棕色脂肪组织。这些发现促使了研究的复苏，该研究集中于棕色脂肪组织的活化和白色脂肪组织的“布朗宁”，这是一种由刺激诱导UCP-1介导的线粒体解偶联的现象。使用啮齿动物模型的研究已经证明，棕色脂肪活化或布朗宁的诱导可以促进有益的代谢作用，并且已经确定了有希望的治疗靶点。然而，由于缺乏广泛可用的人棕色脂肪细胞模型，这些靶点的治疗潜力尚未得到证实。因此，有一个明显的需要，以验证现有的治疗目标在人类系统中。ZenBio将通过向研究界提供原代和永生化的人类棕色脂肪细胞培养系统来满足这一需求，并将利用这些工具来建立筛选以识别人类棕色脂肪生成的激活剂。该项目的主要目标是产生和表征人成人棕色脂肪细胞系统。我们将以逐步的方式实现这一目标，首先是对人类胎儿棕色脂肪细胞进行初步的可行性研究。目的利用功能分析和基因组分析技术建立一个广泛应用的胎儿棕色脂肪细胞模型，为建立成人脂肪细胞系统奠定基础。由于胎儿棕色脂肪组织的检测相对容易，并且可以通过现有的采购机构随时获得，因此将首先开发胎儿模型。第二个目的是产生和表征用于高通量/高含量筛选的永生化胎儿棕色脂肪细胞系统。这些细胞将避开原代细胞的有限材料和寿命，并允许开发高通量筛选平台以鉴定棕色脂肪细胞活化的新靶点。I期将在II期中扩展，目标是表征和商业化来自不同BMI、年龄、性别和糖尿病状态的供体的人成人棕色脂肪细胞系统。这些研究将产生原代和永生化的人成人脂肪细胞，其将用于鉴定用于治疗肥胖症的棕色脂肪细胞能量消耗的新靶点。

 DESCRIPTION (provided by applicant): The activation and recruitment of brown adipose tissue has become an exciting target in the fight against obesity and its related metabolic diseases. Brown adipose tissue differs from white adipose tissue in its critical ability to burn energy as heat through UCP-1 driven adaptive thermogenesis. Expression of mitochondrial UCP-1 uncouples oxidative phosphorylation, resulting in accelerated substrate oxidation but low ATP production. The resulting dissipation of energy is associated with plasma triglyceride clearance, a key factor in weight loss. The brown adipose depot has long been known to exist in rodents as well as in human infants. However, recent evidence has demonstrated the existence of active brown adipose tissue in human adults. These findings prompted a resurgence of research focusing on both the activation of brown adipose tissue and the "browning" of white adipose tissue, a phenomenon wherein UCP-1-mediated mitochondrial uncoupling is induced by stimuli. Studies using rodent models have demonstrated that brown fat activation or induction of browning can promote beneficial metabolic effects, and promising therapeutic targets have been identified. However, the therapeutic potential of these targets has not been confirmed due to the lack of a widely available human brown adipocyte model. Therefore, there is a pronounced need to validate existing therapeutic targets in the human system. ZenBio will address this need by providing both a primary and immortalized human brown adipocyte cell culture system to the research community and will utilize these tools to establish a screen to identify activators of human brown adipogenesis. The principal goal of this project is to generate and characterize a human adult brown adipocyte cell system. We will achieve this in a stepwise fashion, beginning with an initial feasibility study focused on human fetal brown adipocytes. Aim I will use functional and genomic analyses to generate a widely available fetal brown adipocyte model which will be used as a tool to establish the adult cell system. The fetal model will be developed first due to the comparative ease of detecting fetal brown adipose tissue and its ready availability through existing procurement agencies. The second aim is to generate and characterize an immortalized fetal brown adipocyte system for high throughput/high content screening. These cells will circumvent the limited material and lifespan of primary cells and allow for the development of a high throughput screening platform to identify novel targets of brown adipocyte activation. Phase I will be expanded in Phase II with the goal of characterizing and commercializing the human adult brown adipocyte system from donors of differing BMI, age, gender and diabetic state. These studies will generate both primary and immortalized human adult adipocyte cells which will be used to identify novel targets of brown adipocyte energy expenditure for the treatment of obesity.