58 Single-cell comparative analysis reveals a similar regulatory subpopulation in white and brown adipocyte precursors

Abstract

Objectives/Goals: The goal of this study is to resolve the complexity of the adipose precursor cells and identify potential therapeutic targets/mechanism to treat obesity, diabetes, cancer cachexia, and related metabolic conditions in both white and brown adipose tissues. Methods/Study Population: White and brown adipose precursor cells were isolated from neonatal P0 mice and expanded in culture for single-cell RNA sequencing analysis. Unsupervised machine learning was used to unbiasedly cluster and categorize sequenced cells. Differentially expressed genes were used to identify populations via pathway analysis. Populations were then validated with published datasets via integration, reference mapping, and module scoring to ensure our dataset is reflective of known literature. Then, white and brown datasets were combined and unbiasedly clustered. Finally, signaling inferences using CellChat was used to identify significant signals being sent to and received from each cluster based on ligand–receptor pairs. Results/Anticipated Results: ScRNAseq revealed 7 subclusters in both white and brown adipose tissues. Differential expression and trajectory inferences revealed that white and brown precursors develop into two distinct fates: committed adipogenic precursors (CAPs), where these cells will be mature lipid-laden adipocytes; or fibro-adipogenic precursors-like (FAPLs), where these cells preferably stay in a fibroblast-like, antiadipogenic phenotype. Integrating white and brown cells with subsequent reclustering reveals that white and brown FAPLs are highly similar to one another by being clustered together. Cell signaling inferences and pathway analysis reveal that white and brown FAPLs may participate in the regulation of adipogenesis and angiogenesis of the adipose tissue. Discussion/Significance of Impact: Our results demonstrated that brown and white precursor cells share a common regulatory subpopulation with similar gene expression profiles, highlighting a more interconnected regulatory landscape in adipose tissue than previously understood. These findings reveal novel mechanisms of systemic metabolism and provide new therapeutic targets.