Sex‑specific cardiovascular risk in estrogen‑treated androgen‑deprived males: metabolic characterization of glucose, adipose, and lipid pathways

Abstract Background Estrogen therapy and androgen‑deprivation were once combined to treat prostate cancer (PrCa). Clinical studies later showed that prolonged estrogen exposure in androgen‑deprived men raises cardiovascular disease (CVD) risk, yet the metabolic pathways responsible remain unclear. Methods We generated an androgen‑deprived, 17β‑estradiol (E2)–treated mouse model by gonadectomizing male C57BL/6 J mice and implanting sub‑cutaneous delayed‑release E2 or vehicle pellets. Mice received a Western‑style diet and were housed at thermoneutrality to accelerate CVD‑risk phenotypes. Metabolic profiling included hyperinsulinemic‑euglycemic clamps, oral lipid and pyruvate tolerance tests, flow cytometry of immune cells, and single‑nucleus RNA sequencing of liver tissue. Results In hypogonadal males, E2 treatment induced several metabolic disturbances. During clamps, E2‑treated mice showed markedly elevated gluconeogenesis, corroborated by higher glucose peaks and AUC during pyruvate tolerance testing and by up‑regulation of hepatic Pck1 mRNA. Triglyceride (TG) clearance, which improves with E2 in females, was impaired in E2‑treated males: oral lipid‑tolerance testing revealed prolonged TG excursions, reduced maximal lipase activity, lower non‑lipase clearance at 6 h post-OLTT, and decreased free‑fatty‑acid peak levels. Hepatic lipase, VLDL clearance receptors Ldlr and Lrp1, and microsomal triglyceride transfer protein (MTP) transcripts were down‑regulated. SnRNA‑seq showed suppression of lipid‑clearance genes with E2 treatment in males. Subcutaneous adipocytes were hypertrophic, and flow cytometry identified increased TNFα‑positive macrophages, an inflammatory milieu that could promote insulin resistance. Cardiac morphology was modestly altered; E2‑treated males exhibited a larger left‑ventricular end‑diastolic diameter, while ejection fraction and arterial pressure remained unchanged. Conclusion Estradiol administration in androgen‑deprived male mice produces a constellation of metabolic derangements—including enhanced hepatic gluconeogenesis, impaired TG clearance, and inflammatory adipocyte hypertrophy—that likely underlie the increased CVD risk observed clinically. The identified molecular nodes (PEPCK, hepatic lipase, LRP1, LDLR, MTP, and adipose‑macrophage TNFα) provide potential targets for mitigating estrogen‑induced CVD risk while preserving its therapeutic benefit for PrCa. Graphical abstract