Investigating the regulation of distinct human adipocyte subpopulations

项目摘要 脂肪组织是能量平衡和葡萄糖稳态的中心参与者，它能够在面对 卡路里超载，以便安全储存能量，但它可能变得超载和功能失调，导致 以胰岛素抵抗和2型糖尿病为形式的全身代谢损害。调查差异 在瘦子和肥胖者的单个细胞类型中，我进行了单核RNA测序(SNUC- (SEQ)在人类皮下和内脏白色脂肪组织上进行了研究，并创建了存在于 白色脂肪组织。这项工作的一个主要发现是确定了不同的亚群 脂肪细胞，其中一些与体重指数(BMI)有关。通过将我们的数据与基因组相关联- 对于T2D等代谢特征的广泛关联研究(GWAS)，我们还预测一些脂肪细胞 亚群与代谢性疾病有关。该项目的目标是确定以下因素 使一个亚群倾向于另一个亚群，在外部和转录上都是如此。为了做到这一点，我将表演 SNUC-SEQ在减肥手术中和术后收集的受试者脂肪组织上的SNUC-SEQ 减肥过程中脂肪细胞亚群的变化。接下来我将询问脂肪细胞的染色质状态 脂肪单核转座酶可及染色质分析的亚种形成 瘦人和肥胖者的组织。最后，我将直接测试潜在的信号和转录调节因子 通过执行潜在信号调节因子的筛选以及CRISPRa筛选来确定亚群身份 潜在的转录调控因子，试图在体外概括不同的亚群。我的经历是 在单细胞分辨率下表征脂肪组织方面，以及我的导师和合作伙伴的经验 研究脂肪细胞转录和基因调控的导师使我处于独特的位置来回答 这些问题。综上所述，这些研究将增强我们对人类白色脂肪细胞多样性的了解 并将在我自己的独立实验室和整个社区为下游研究奠定基础。

Project Summary Adipose tissue is a central player in energy balance and glucose homeostasis, it is able to expand in the face of caloric overload in order to store energy safely, but it can become overloaded and dysfunctional, leading to systemic metabolic compromise in the form of insulin resistance and Type 2 diabetes. To investigate differences in individual cell types in lean and obese individuals, I have performed single nucleus RNA sequencing (sNuc- seq) on human subcutaneous and visceral white adipose tissue and created an atlas of cell types present in white adipose tissue. A major finding from this work was the identification of distinct subpopulations of adipocytes, some of which are associated with body mass index (BMI). By associating our data with genome- wide association studies (GWAS) for metabolic traits such as T2D, we additionally predict that some adipocyte subpopulations are associated with metabolic disease. The objective of this project is to identify factors that predispose one subpopulation over another, both externally as well as transcriptionally. To do this I will perform sNuc-seq on adipose tissue collected from subjects during and post-bariatric surgery in order to characterize the change in adipocyte subpopulation during weight loss. I will next interrogate the chromatin state of adipocyte subpoulations by performing the Assay for Transposase Accessible Chromatin on single nuclei from adipose tissue of lean and obese individuals. Finally, I will directly test potential signaling and transcriptional regulators of subpopulation identity by performing a screen of potential signaling regulators as well as a CRISPRa screen of potential transcriptional regulators to try to recapitulate distinct subpopulations in vitro. The experience that I have in characterizing adipose tissue at single cell resolution, as well as the experience of my mentor and co- mentor in studying transcriptional and genetic regulation of adipocytes make me uniquely positioned to answer these questions. Taken together, these studies will enhance our knowledge of human white adipocyte diversity and will set the stage for downstream studies in my own independent lab and in the community at large.