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

研究脂肪组织细胞异质性的新型遗传工具的产生

基本信息

批准号：
9816331
负责人：
Rana K Gupta
金额：
$ 144.06万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-17 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9816331
关键词：
Adipocytes Adipose tissue Adopted Adult Anatomy Animal Model Area Atlases Attention Biology Brown Fat Cell Lineage Cells Collection Communities Complex Data Databases Development Energy Metabolism Enterobacteria phage P1 Cre recombinase Event Gene Expression Gene Expression Profile Gene Targeting Generations Genes Genetic Genetic Recombination Goals Heterogeneity Homeostasis Individual Inflammatory Infrastructure Internal Medicine Joints Laboratories Logistics Mediating Metabolism Methodology Mission Modeling Molecular Mouse Strains Mus National Institute of Diabetes and Digestive and Kidney Diseases Nutrient Pericytes Physiological Physiology Population Positioning Attribute Promoter Regions Protein Splicing Publications Recording of previous events Research Research Personnel Resources Specificity Stromal Cells System Technology Time Tissues Visceral base cell type cellular targeting genetic approach innovation intein interest mature animal molecular subtypes mouse model next generation novel phenotypic data pre-clinical precursor cell progenitor promoter reconstitution single cell sequencing single-cell RNA sequencing subcutaneous tool

项目摘要

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.

研究脂肪组织细胞异质性的新型遗传工具的产生近年来，人们对细胞有更多的关注在临床前环境中，直接参与维持能量稳态的能力。 Cre 重组酶在特定组织中的表达是一项重大进展。当这些基于 Cre 的重组事件变得可诱导时，Cre 活性暂时且参与该提案的两名研究人员（Gupta 和）在成年动物模型中进行了空间控制。 Scherer）一直处于小鼠模型生成的最前沿，目前广泛应用于新陈代谢领域，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议进行脂肪组织和细胞类型特异性基因操作随着单细胞测序方法变得更加普遍和广泛使用，它已经进入了一个新的水平。越来越明显的是，存在很大程度的细胞异质性，即使是在细胞群体中长期以来被认为是同质的，有强有力的证据表明这种异质基因表达模式。反映了重要的功能差异，特别是在前脂肪细胞和脂肪细胞区域。使用“Split-Cre”开发新型遗传工具来研究脂肪组织中的细胞异质性。方法论，我们将协调努力，将酶 Cre 活性引导至细胞的特定子集该应用程序建立在现有的基因表达数据库的基础上。由两名研究人员产生，并建议扩大努力寻找更多独特或高度我们将利用我们的相应脂肪库和细胞亚群中的富集标记。集体专业知识和机构基础设施，以验证拟议鼠标的功效和特异性这些产生的小鼠模型将构成新的社区资源，并将造福所有脂肪人群。能量代谢一般领域的组织生理学家作为证明证据的额外步骤。这些新模型的概念和功效，我们将使用这些新生成的小鼠来强调选定的细胞亚群的生理相关性。该应用程序的支持将构成一种新资源，可供整个社区使用以类似于我们使用之前生成的遗传工具所做的方式，以及详细的表型数据、表达数据和所有其他相关信息，以实现高效和广泛的使用这些新工具鼠标。