Single-cell RNA sequencing (scRNA-seq) has revealed that adult white adipose tissue (WAT) harbors functionally diverse subpopulations of mesenchymal stromal cells that differentially impact tissue plasticity. To date, the molecular basis of this cellular heterogeneity has not been fully defined. Here, we describe a multilayered omics approach to dissect adipose progenitor cell heterogeneity in three dimensions: progenitor subpopulation, sex, and anatomical localization. We applied state-of-the-art mass spectrometry methods to quantify 4,870 proteins in eight different stromal cell populations from perigonadal and inguinal WAT of male and female mice and acquired transcript expression levels of 15,477 genes using RNA-seq. Our data unveil molecular signatures defining sex differences in preadipocyte differentiation and identify regulatory pathways that functionally distinguish adipose progenitor subpopulations. This multilayered omics analysis, freely accessible at http://preadprofiler.net/, provides unprecedented insights into adipose stromal cell heterogeneity and highlights the benefit of complementary proteomics to support findings from scRNA-seq studies.

单细胞 RNA 测序 (scRNA-seq) 表明，成人白色脂肪组织 (WAT) 具有功能多样的间充质基质细胞亚群，这些亚群对组织可塑性有不同的影响。迄今为止，这种细胞异质性的分子基础尚未完全确定。在这里，我们描述了一种多层组学方法，从三个维度剖析脂肪祖细胞的异质性：祖细胞亚群、性别和解剖定位。我们应用最先进的质谱法对来自雄性和雌性小鼠性腺周围和腹股沟 WAT 的八个不同基质细胞群中的 4,870 种蛋白质进行了量化，并使用 RNA-seq 获得了 15,477 个基因的转录表达水平。我们的数据揭示了定义前脂肪细胞分化性别差异的分子特征，并确定了在功能上区分脂肪祖细胞亚群的调节途径。这种多层组学分析可在 http://preadprofiler.net/ 免费访问，提供了对脂肪基质细胞异质性的前所未有的见解，并强调了互补蛋白质组学支持 scRNA-seq 研究结果的好处。