In-Depth Analysis of Obesity-Associated Changes in Adipose Tissue-Derived Mesenchymal Stromal/Stem Cells and Primary Cilia Function

Adipose tissue-derived mesenchymal stromal/stem cells (ASCs) possess regenerative potential. Obesity induces a pro-inflammatory environment that compromises their function. We investigated how obesity affects ASC biology, focusing on primary cilia. Our data reveal that obesity alters ASC gene expression, particularly in pathways related to the extracellular matrix, transforming growth factor-ß (TGFß) signaling, cell motility, and differentiation. The gene levels of regulatory factor X2 (RFX2) and adenylate cyclase 3 (ADCY3), important for ciliary biogenesis, were downregulated in obese ASCs. TGFß treatment significantly decreases the expression of RFX2 and ADCY3 in lean ASCs. Knockdown of ADCY3 reduced primary cilium length, whereas pharmacological activation restored it and improved cell motility. These results suggest that obesity impairs ASC ciliary function, contributing to defective adipogenesis and reduced regenerative capacity. Restoring ADCY3 activity partially rescues ciliary integrity and cellular function, highlighting the role of primary cilia in ASC dysfunction and offering potential therapeutic targets for obesity-related disorders. Overall design: Adipose tissue-derived mesenchymal stromal/stem cells (ASC) were isolated from subcutaneous (sub) adipose tissue samples from women undergoing cesarean section. Total RNA was extracted from five individual ASC samples per group: lean (body mass index, BMI < 25, SUB25), obese 35 (BMI = 35, SUB35), and obese 40 (BMI = 40, SUB40), and subjected to whole-genome RNA sequencing (RNA-seq).