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：
10617192
负责人：
Rana K Gupta
金额：
$ 134.04万
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 2024-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10617192
关键词：
Adipocytes Adipose tissue Adopted Adult Anatomy Animal Model Area Attention Biology Brown Fat Categories Cell Lineage Cells Collection Communities Complex Cre driver Data Dedications Development Energy Metabolism Enterobacteria phage P1 Cre recombinase Event Gene Expression Gene Expression Profile Gene Targeting Generations Genetic Genetic Recombination Goals Heterogeneity Homeostasis Individual Infrastructure Institution 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 Tissue atlas Tissues Visceral cell type cellular targeting efficacy validation gene expression database genetic approach genetic manipulation 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.

研究脂肪组织中细胞异质性的新型遗传工具近年来，人们一直致力于发展对细胞的更好理解重要的类型参与维持能量稳态。在临床前的环境中，指导 CRE重组酶对特定组织的表达已成为一个重大进展。额外的进步是当这些基于CRE的重组事件的诱导性时，将暂时进行CRE活性和在成人动物模型中得到空间控制。参与该提案的两名调查人员（Gupta和 Scherer）一直处于现在广泛用于代谢中的鼠标模型的最前沿领域，在每种情况下都利用各自脂肪组织中各自感兴趣的细胞类型的专家。现在的两个研究者现在建议采用脂肪组织和细胞类型特异性基因操纵到一个下一个级别。随着单细胞测序方法变得越来越普遍且广泛使用，它是越来越明显的是，即使在细胞种群中，也存在大量的细胞异质性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重要的功能差异，特别是在脂肪细胞和脂肪细胞的领域。我们建议使用“拆分” 方法论，我们将协调将酶促活性引导到细胞特定子集的努力不同脂肪组织中的种群。此应用程序建立在现有基因表达数据库的基础上由两个调查人员产生，以及扩大努力以寻找其他独特或高度的提议相对脂肪沉积物和细胞亚群中的富集标记。我们将利用我们的集体专业知识和机构基础设施，以验证拟议的鼠标的效率和特异性菌株。这些生成的鼠标模型将构成一个新的社区资源，并将受益于所有脂肪能量代谢一般领域的组织生理学家。作为展示证明的额外步骤这些新模型的概念和便捷性，我们将使用这些新生成的小鼠强调细胞选定亚群的生理相关性。这些鼠标模型在该应用程序的主持将构成一种新资源，将为整个社区提供以类似于我们以前生成的遗传工具所做的类似的方式表型数据，表达数据和所有其他相关性信息，以允许有效而宽度使用这些新工具小鼠。