Inhibition of the NR4A1‐mediated cell cycle checkpoint enhances hyperplastic adipogenesis and glucose homeostasis in obesity

BACKGROUND AND PURPOSE: The nuclear receptor NR4A1 is a critical node in transcriptional circuits for metabolic adaptation, yet its role in adipocyte progenitor (AP) fate and proliferation is unclear. We sought to define how NR4A1 governs early adipogenesis and adipose tissue plasticity. EXPERIMENTAL APPROACH: Doxycycline-inducible NR4A1-overexpressing and NR4A1-deficient 3T3-L1 models were used to assess lipid accumulation, mitotic clonal expansion (MCE), and cell-cycle dynamics. Multiomics integration (RNA-seq and chromatin assays) identified NR4A1 targets. The functionality of two lead candidates, ANLN and CDC20, was tested by genetic knockdown and pharmacological inhibition. Diet-induced obese mice received the NR4A1 antagonist C-DIM8 alone or combined with the PPARγ agonist rosiglitazone to evaluate adipose expansion and metabolic outcomes. KEY RESULTS: NR4A1 exerted a stage-specific inhibitory effect during early differentiation, reducing expression of canonical adipogenic markers and lipid accumulation. Integrative transcriptomic and chromatin analyses identified Anln and Cdc20 as direct NR4A1 targets essential for MCE. Depletion of either gene phenocopied NR4A1 overexpression, inducing G2/M arrest and blunting of adipogenesis. Pharmacological CDC20 inhibition with Apcin suppressed preadipocyte proliferation and differentiation in a dose-dependent manner. Importantly, in a diet-induced obese mouse model, combined treatment with the NR4A1 antagonist C-DIM8 and the PPARγ agonist rosiglitazone promoted AP proliferation, hyperplastic adipose tissue expansion, and improved glucose tolerance. CONCLUSIONS AND IMPLICATIONS: NR4A1 functions as a transcriptional gatekeeper of adipogenic initiation by restraining the Anln/Cdc20 cell-cycle program. Pharmacological modulation of the NR4A1-cell-cycle axis, particularly alongside PPARγ activation, may offer a strategy to augment adipose tissue plasticity and metabolic resilience in obesity.