Single-nucleus RNA Sequencing Reveals Inguinal White Adipose Tissue Remodeling Upon Return to Thermoneutrality After Cold Stimulation

Cold exposure induces browning of inguinal white adipose tissue (iWAT), making it a prime target for therapies addressing obesity and metabolic disorders. However, this remodeling is reversible when returning to thermoneutrality, iWAT whitens and loses its enhanced metabolic function. To investigate this dynamic, we first established returning to thermoneutrality for 2 weeks as a critical cut-off point for iWAT remodeling. Single-nucleus RNA sequencing (SnRNAseq) was then employed to map the cellular landscape of iWAT throughout the entire process, from cold exposure to rewarming. This comprehensive altas encompassed adipocytes, adipose stem and progenitor cells (ASPCs), immune cells, endothelial cells, and more, providing a detailed view of transcriptomic and intercellular interaction dynamics. Our findings revealed that despite a substantial reversion to the initial whitened state after rewarming, including structural features, Cellular changes and functional responses, some cold-induced effects persisted. Specifically, ASPCs returning to warm environments exhibited altered differentiation patterns, suggestive of a "de-differentiation" trend. Furthermore, the ANGPTL pathway, activated in thermogenic adipocyte subpopulation A3 during cold exposure, remained involved in intercellular communication even after whitening and loss of thermogenic capacity. This suggests a potential role for ANGPTL signaling in maintaining the whitened phenotype and regulating iWAT plasticity.