Generation of Novel Genetic Tools to Study Cellular Heterogeneity in Adipose Tissues

脂肪组织细胞异质性研究的新遗传工具的产生 近年来，人们一直致力于更好地了解细胞 与维持能量平衡密切相关的类型。在临床前环境中， Cre重组酶在特定组织中的表达是一个重大进展。另一项进展是， 当这些基于Cre的重组事件是可诱导的时，Cre的活性在时间上是可诱导的， 在成年动物模型中进行空间控制。参与这项提案的两位调查人员（古普塔和 谢勒）一直处于小鼠模型产生的最前沿，现在广泛用于代谢 领域，在每种情况下利用不同脂肪组织中各自感兴趣的细胞类型的专业知识。 这两位研究人员现在提出，对脂肪组织和细胞类型进行特异性基因操作， 更上一层楼。随着单细胞测序方法变得越来越普遍和广泛使用， 越来越明显的是，存在着巨大程度的细胞异质性，即使在细胞群体中， 长期被认为是同质的。有强有力的证据表明，这种异质性的基因表达模式 反映了重要的功能差异，特别是在前脂肪细胞和脂肪细胞领域。我们提出 开发新的遗传工具来研究脂肪组织中的细胞异质性。使用“Split-Cre” 方法，我们将协调努力，直接酶Cre活性的特定子集的细胞 在不同的脂肪组织中。该应用程序建立在现有的基因表达数据库上 产生的两个调查员，并建议扩大努力，寻找更多的独特或高度 在各自的脂肪库和细胞亚群中富集标志物。我们将利用我们 集体专业知识和机构基础设施，以验证拟议的鼠标的功效和特异性 菌株这些产生的小鼠模型将构成一个新的社区资源，并将有利于所有脂肪 能量代谢领域的组织生理学家。作为一个额外的步骤来证明- 这些新模型的概念和功效，我们将使用这些新产生的小鼠来强调 选择的细胞亚群的生理相关性。这些小鼠模型是在 这项应用程序的赞助将构成一个新的资源，将提供给广大社区 以类似于我们以前生成的遗传工具所做的方式，沿着详细的 表型数据、表达数据和所有其他相关信息，以允许高效和广泛的 使用这些新的工具鼠标。

Generation of Novel Genetic Tools to Study Cellular Heterogeneity in Adipose Tissues Much attention has been dedicated over recent years towards developing a better understanding of the cell types critically involved in maintaining energy homeostasis. In preclinical settings, the ability to direct the expression of Cre recombinase to specific tissues has been a major advance. An additional advance was made when these Cre-based recombination events were made inducible, with Cre activity temporally and spatially controlled in adult animal models. The two investigators involved in this proposal (Gupta and Scherer) have been at the forefront of the generation of mouse models now widely used in the metabolism field, exploiting in each case the expertise in their respective cell types of interest in different adipose tissues. The same two investigators now propose to take the adipose tissue- and cell type specific gene manipulation to the next level. With single cell sequencing approaches becoming more commonplace and widely used, it is increasingly obvious that a vast degree of cellular heterogeneity exists, even in cellular populations that were long considered homogeneous. There is strong evidence that such heterogeneous gene expression patterns reflect important functional differences, particularly in the areas of pre-adipocytes and adipocytes. We propose the development of novel genetic tools to study cellular heterogeneity in adipose tissues. Using “Split-Cre” methodology, we will coordinate efforts to direct enzymatic Cre activity to specific subsets of cellular populations in different adipose tissues. This application builds upon existing gene expression databases generated by the two investigators, and proposes to expand the efforts in finding additional unique or highly enriched markers in the respective adipose depots and the cellular subpopulations. We will leverage our collective expertise and institutional infrastructure to validate the efficacy and specificity of the proposed mouse strains. These mouse models generated will constitute a new community resource and will benefit all adipose tissue physiologists in the general field of energy metabolism. As an additional step to demonstrate proof-of- concept and efficacy of these new models, we will use these newly generated mice to underline the physiological relevance of selected subpopulations of cells. These mouse models generated under the auspices of this application will constitute a new resource that will be made available to the community at large in a manner similar to what we have done with our previously generated genetic tools, along with detailed phenotypic data, expression data, and all other information of relevance, to allow for efficient and widespread use of these new tool mice.