Zfp423 in the Control of Preadipocyte Determination

描述（由申请人提供）：肥胖症的全球流行令人担忧，因为肥胖症是许多慢性疾病（如2型糖尿病、癌症和心血管疾病）的重要风险因素。操纵脂肪谱系可能代表肥胖和相关疾病的新的治疗方法;然而，任何尝试都将取决于对控制脂肪细胞发育的基本机制的更深入理解。我的职业目标是成为新兴的脂肪发育领域的领先独立研究者，特别关注控制体内不同脂肪库初始规格的发育机制。到目前为止，我在达纳-法伯癌症研究所布鲁斯斯皮格曼博士实验室的博士后研究集中在发现前脂肪细胞决定的转录调节因子上。我们最近发现的转录因子Zfp 423作为前脂肪细胞富集的前脂肪细胞定型的调节因子，对体外和体内脂肪细胞发育至关重要，这为1）开发定位和分离体内发现的定型前脂肪细胞的工具，以及2）阐明控制脂肪谱系定型的分子机制开辟了可能性。在具体目标1中，我们将利用表达GFP的BAC转基因小鼠在Zfp 423顺式调节元件的控制下定位和分离各种成年脂肪库以及发育中的小鼠胚胎中的前脂肪细胞，并证明它们对脂肪谱系的承诺。在具体目标2中，我们将通过Zfp 423功能和相互作用伙伴的生化分析来阐明前脂肪细胞决定的分子机制。这些实验将揭示在小鼠发育过程中定向前脂肪细胞的解剖位置，建立一种新的工具，用于纯化小鼠脂肪前体，确定Zfp 423控制前脂肪细胞决定的分子机制，重要的是，确定前脂肪状态的新调节剂。对这些未探索的脂肪生物学领域的更深入理解将有助于我们尝试操纵脂肪谱系作为肥胖和代谢疾病的治疗方法。这项职业发展奖的支持以及开展这些实验所获得的经验将促进我转变为脂肪生物学领域的独立研究者。 公共卫生相关性：肥胖症发病率的上升增加了了解控制能量稳态的基本系统的紧迫性，包括脂肪组织的形成。本实验将为小鼠脂肪细胞前体的研究提供重要的工具，并揭示脂肪细胞发育的分子机制。这些知识将有助于我们尝试操纵脂肪组织生物学作为肥胖和代谢疾病的治疗方法。

DESCRIPTION (provided by applicant): The global epidemic of obesity is alarming, as obesity is a significant risk factor for numerous chronic disorders such as type 2 diabetes, cancer, and cardiovascular disease. Manipulating the adipose lineage may represent a novel therapeutic treatment for obesity and associated diseases; however, any attempts will depend on a deeper understanding of the basic mechanisms controlling adipocyte development. My career goal is to become a leading independent investigator in the burgeoning field of adipose development, with particular focus on the developmental mechanisms controlling the initial specification of different adipose depots in vivo. My postdoctoral research to date in Dr. Bruce Spiegelman's laboratory at the Dana-Farber Cancer Institute has focused on the discovery of transcriptional regulators of preadipocyte determination. Our recent discovery of the transcription factor Zfp423 as a preadipocyte-enriched regulator of preadipocyte commitment crucial for adipocyte development in vitro and in vivo has opened up the possibility of 1) developing tools to localize and isolate committed preadipocytes found in vivo, and 2) elucidate the molecular mechanisms controlling commitment to the adipose lineage. In Specific Aim 1 we will utilize BAC transgenic mice expressing GFP under the control of Zfp423 cis-regulatory elements to localize and isolate preadipocytes in various adult fat depots as well as in developing mouse embryos, and demonstrate their commitment to the adipose lineage. In Specific Aim 2 we will elucidate the molecular mechanisms of preadipocyte determination through biochemical analysis of Zfp423 function and interacting partners. These experiments will reveal the anatomical location of committed preadipocytes during mouse development, establish a novel tool for the purification of adipose precursors in mice, determine the molecular mechanisms by which Zfp423 controls preadipocyte determination, and importantly, identify novel regulators of the preadipose state. A deeper understanding of these unexplored areas of adipose biology will facilitate our attempts to manipulate the adipose lineage as a therapeutic treatment for obesity and metabolic disease. The support of this career development award and the experience derived from carrying out these experiments will facilitate my transition to an independent investigator in the field of adipose biology. PUBLIC HEALTH RELEVANCE: The rising incidence of obesity has increased the urgency of understanding the fundamental systems controlling energy homeostasis, including the formation of fat tissue. The experiments described in this proposal will establish and utilize an important tool for studying fat cell precursors in mice, and reveal the molecular mechanisms controlling fat cell development. This knowledge will facilitate our attempts to manipulate fat tissue biology as a therapeutic treatment for obesity and metabolic disease.