Exploring Potential Multi-Target Inhibition Mechanisms of Ganoderma lucidum Against EBV-Mediated DLBCL Using...

Epstein-Barr virus (EBV)-associated diffuse large B-cell lymphoma (DLBCL) represents a significant clinical challenge with limited targeted therapeutic options. Despite the known antiviral and anticancer properties of Ganoderma lucidum, its specific molecular mechanisms against EBV-mediated DLBCL remain unexplored. Therefore, this study aimed to investigate the potential multi-target inhibitory mechanisms of G. lucidum compounds against EBV-associated DLBCL by integrating absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiling of 1046 compounds; network pharmacology analysis for target identification; transcriptomic evaluation using DLBCL patient datasets from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) (n = 47 tumors, n = 337 normals); molecular docking; density functional theory (DFT) calculations; machine learning-based activity prediction, molecular dynamics (MD) simulations; and Molecular Mechanics Poisson-Boltzmann Surface Area (MM-PBSA) binding free energy calculations. Eight compounds (kobusin, chrysin, dalbergin, galangin, isorhamnetin, lauric acid, piperbetol, and vitamin D3) demonstrated favorable ADMET profiles. Network pharmacology analysis identified 217 common targets, with AKT1 and BCL2 significantly upregulated in DLBCL tumors. Isorhamnetin showed the strongest AKT1 binding (-8.33 kcal/mol), surpassing control (-5.11 kcal/mol), whereas galangin exhibited the highest BCL2 affinity (-6.13 kcal/mol), outperforming control (-5.23 kcal/mol). Random forest achieved cross-validation scores of 0.8634 ± 0.0158 for AKT1 and 0.8965 ± 0.0146 for BCL2, predicting IC₅₀ <1000 nM for all compounds. MD simulations confirmed the binding stability of all compounds with low root mean square deviation (RMSD, <2.5 Å). MM-PBSA calculations revealed favorable binding free energies of -23.01 kcal/mol (AKT1-galangin), -19.01 kcal/mol (AKT1-isorhamnetin), -32.15 kcal/mol (BCL2-isorhamnetin), and -28.97 kcal/mol (BCL2-galangin). Therefore, isorhamnetin and galangin may have multi-target inhibitory potential against AKT1 and BCL2, providing a robust foundation for further in vitro and in vivo validation as therapeutic candidates for EBV-mediated DLBCL intervention. PRACTICAL APPLICATIONS: Ganoderma lucidum is a medicinal mushroom widely used in traditional and functional food applications and may serve as a source of natural anticancer compounds with potential benefits for EBV-associated diffuse large B-cell lymphoma. The multi-targeting potential of G. lucidum-derived bioactive constituents, isorhamnetin and galangin, against AKT1 and BCL2 may position it as a promising functional food-based candidate for anticancer therapeutics.