DHODH regulates mitochondrial bioenergetics, methylation cycle, and DNA repair in Prostate Adenocarcinoma

African American (AA) men exhibit disproportionately high incidence rates of prostate cancer (PCa) and a greater propensity for aggressive disease progression. Although epidemiological studies have delineated various risk factors, the precise molecular mechanisms driving the unique tumor biology and clinical disparities in AA men remain poorly characterized. Our prior investigations employing in situ analyses of clinical PCa tissues revealed increased uracil incorporation and elevated pyrimidine damage, indicative of heightened DNA repair activity, particularly within AA prostate tumours. Notably, dihydroorotate dehydrogenase (DHODH), a critical enzyme involved in de novo pyrimidine biosynthesis, was significantly overexpressed in PCa, specifically in epithelial cells, with the highest expression observed in patient-derived tumours from AA patients. Through comprehensive gain and loss-of-function experiments across multiple PCa cell lines integrating transcriptomic profiling, DNA methylation sequencing, and metabolomics we demonstrate that DHODH centrally orchestrates mitochondrial bioenergetics, one-carbon metabolism, methylation pathways, and DNA repair mechanisms. Functionally, modulation of DHODH expression directly influenced cellular proliferation, migration, and metastatic potential in vivo, accompanied by alterations in base excision repair (BER)-mediated DNA damage. Collectively, our findings position DHODH as a pivotal metabolic and epigenetic regulator with critical implications for PCa aggressiveness and racial disparities. This work establishes DHODH as a promising therapeutic target, potentially addressing unmet clinical needs and improving outcomes specifically for AA prostate cancer patients.