Abstract B033: Proteomic and i <i>n silico</i> insights reveal novel targets of β-hydroxybutyrate in prostate cancer cells

Abstract While β-hydroxybutyrate (BHB), a predominant ketone body, has shown anti-cancer activity in various malignancies, its molecular interactions and genomic effects in prostate cancer (PCa) remain largely unexplored. This study employs integrative proteomics and in silico modeling to delineate the mechanisms by which BHB modulates prostate cancer cell signaling, particularly in LNCaP cells under lipopolysaccharide (LPS)-stimulated stress. Following treatment with BHB and LPS, LNCaP cells underwent high-throughput proteomic profiling. Differentially expressed proteins were analyzed through protein-protein interaction (PPI) networks using the CytoHubba plug-in. Functional enrichment was conducted via ShinyGO, and five key genes—acireductone dioxygenase 1 (ADI1), ubiquitin-specific peptidase 54 (USP54), succinate-CoA ligase alpha subunit (SUCLA2), ubiquitin-specific peptidase 16 (USP16), and heterogeneous nuclear ribonucleoprotein H1 (HNRNPH1)—were subjected to molecular docking simulations with BHB. Proteomic results revealed 256 significantly altered genes, of which 146 were downregulated upon BHB + LPS co-treatment. Among the top 20 hub genes, ADI1 was uniquely upregulated, suggesting a stress-adaptive or detoxifying role. Gene ontology (GO) enrichment identified overrepresented pathways in transcriptional elongation, innate immune response, and aspartate biosynthesis. Subcellular localization analysis showed that these processes predominantly occur in mitochondria, cytoplasm, and ribosomes, implicating BHB in cellular energy and immune reprogramming. In silico docking further highlighted a high-affinity interaction between BHB and USP54 (binding affinity: −6.85 kcal/mol; complex stability: −25.92 kcal/mol), a gene associated with cancer invasion and immune escape. Docking results for USP16, SUCLA2, and HNRNPH1 suggested these proteins could also act as downstream effectors in BHB-mediated signaling. Collectively, the integration of omics data and computational modeling provides novel insight into the gene networks and targets affected by BHB. These findings suggest that BHB not only modulates inflammatory and metabolic pathways but also interfaces with key regulatory proteins involved in PCa metastasis and immune modulation. Thus, BHB holds translational potential as a multitargeted metabolic agent in precision oncology for prostate cancer. Citation Format: Omowumi kayode, Olatayo Afolabi, Gabriel Ajayi. Proteomic and in silico insights reveal novel targets of β-hydroxybutyrate in prostate cancer cells [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Innovations in Prostate Cancer Research and Treatment; 2026 Jan 20-22; Philadelphia PA. Philadelphia (PA): AACR; Cancer Res 2026;86(2_Suppl):Abstract nr B033.