Brown adipose tissue (BAT) is a critical target for obesity treatment, and exercise can enhance BAT function through the activation of ADRβ3. However, the molecular mechanisms underlying BAT metabolism following the exercise-induced activation of ADRβ3 remain unclear. This study utilized RNA sequencing, Western blotting, Oil Red O staining, weighted gene co-expression network analysis (WGCNA), and machine learning to investigate the role of the ADRβ3-COX2 pathway in lipid metabolism in brown adipocytes. We identified Ywhah as a key gene and validated our findings using external datasets. Our results demonstrate that exercise significantly enhances brown adipose tissue metabolism in mice, with ADRβ3 activation promoting metabolic activity in brown adipocytes. In contrast, COX2 inhibition notably reduced the lipolytic effect and thermogenic gene expression induced by ADRβ3 activation. WGCNA and machine learning identified Ywhah as the most important feature variable in the downstream signaling of the ADRβ3-COX2 pathway. External microarray data further confirmed that 8 weeks of aerobic exercise significantly upregulated Ywhah expression. Additionally, Ywhah displayed strong binding affinity to cytoskeletal proteins in affinity purification–mass spectrometry experiments, and its expression was highly correlated with cytoskeletal GSVA scores. In summary, this study reveals the potential role of the ADRβ3-COX2-Ywhah-cytoskeleton axis in regulating brown adipocyte metabolism, providing new insights into obesity treatment mechanisms.

Background/Objectives: Anthracycline-based chemotherapy is associated with systemic toxicities in patients with breast cancer. Aerobic exercise may help mitigate treatment-related effects, but its associated metabolic responses during chemotherapy remain poorly understood. This study assessed biochemical and circulating metabolomic responses to aerobic exercise during anthracycline-based chemotherapy. Methods: This exploratory secondary analysis included 40 participants from a randomized controlled trial (trained n = 21; control, n = 19). Participants in the trained group completed a 12-week aerobic exercise intervention during chemotherapy. Biochemical markers and untargeted plasma LC–MS metabolomic profiles were assessed before and after intervention. Exploratory metabolic candidates were evaluated using linear mixed-effects models with Benjamini–Hochberg correction. Results: Significant Group × Time interactions were observed for 10 of 12 physiological and biochemical markers after BH-FDR correction in the primary analysis. However, baseline-adjusted ANCOVA sensitivity analyses showed that only IL-6 retained statistical significance after BH-FDR correction (adjusted between-group difference: −15.58 pg/mL, 95% CI: −20.72 to −10.45; FDR < 0.001). Despite limited global metabolomic separation, sequential screening identified 26 candidate features, seven of which showed significant Group × Time interactions after correction across candidates. Five had putative annotations, including lysophosphatidylcholine (LPC) 14:1, lysophosphatidylethanolamine (LPE) 17:1, cis-7-hexadecenoic acid, L-ascorbate, and vitamin E acetate. In a full-feature sensitivity analysis, only Com_3317_pos remained significant after BH-FDR correction across all 1150 metabolic features (FDR = 0.0228). Conclusions: Aerobic exercise during anthracycline-based chemotherapy was associated with changes in circulating biochemical markers, although these findings were less robust after baseline adjustment. Untargeted metabolomics identified exploratory exercise-associated metabolic features, particularly those putatively related to lipid metabolism, but most did not remain significant after full-feature multiple-testing correction. Further validation is warranted.

Physical inactivity contributes to type 2 diabetes (T2D), but the molecular links between exercise and metabolic improvement remain incompletely understood. We meta-analyzed genome-wide association studies of vigorous physical activity and T2D (combined n ≈ 1.95 million) and integrated eQTL/sQTL maps with single-cell and spatial transcriptomic datasets to connect genetic risk with tissues, cell types, and regulatory programs. Tissue and cell-type enrichment, colocalization, and network analyses were performed. Computational findings were further examined in male 10-week-old C57BL/6J mice with high-fat diet-induced diabetes. After 1 week of acclimatization, mice were randomly assigned to normal chow, high-fat diet, or high-fat diet plus exercise groups (n = 6 per group; high-fat diet with 60% of total energy from fat). The exercise intervention consisted of treadmill running (10 m/min for 50 min per day, 5 days per week, total 16 weeks), followed by metabolic phenotyping, skeletal muscle histology, bulk RNA sequencing, alternative splicing analysis, and RT-qPCR of Mau2 isoforms. Exercise- and T2D-associated variants showed joint enrichment in skeletal muscle and adipose eQTL/sQTL signals. Integrated single-cell analyses prioritized fibro-adipogenic progenitors and endothelial cells, and identified an extracellular matrix- and collagen-related module in fibro-adipogenic progenitors associated with both exercise and T2D. Mau2 emerged as a shared candidate gene with tissue-specific splicing signals. In diabetic mice, exercise improved glucose homeostasis and muscle fiber structure, and reduced Mau2 intron retention in skeletal muscle without changing total Mau2 expression. These findings support a multiscale framework linking exercise-responsive regulation to T2D-related tissue remodeling and splicing plasticity.