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Genome-Wide CRISPR/Cas9 Screening Reveals Lipid Metabolism and Inflammatory Signalling as Modulators of Ganoderic Acid DM Cytotoxicity

Abdullah, N.; Lewis, J.; Arumugam, P.

2025-10-15 genomics
10.1101/2025.10.14.682027 bioRxiv
Show abstract

Ganoderic Acid DM (GA-DM), a triterpenoid derived from Ganoderma lucidum, exhibits anti-cancer and anti-diabetic activities, but the underlying mechanisms of action remain unclear. To identify genetic modulators of GA-DM response, we conducted a genome-wide CRISPR/Cas9 knockout screen in human melanoma cells. The screen revealed key roles for genes regulating lipid metabolism and inflammatory signalling, particularly the SREBP (Sterol Regulatory Element-binding Protein) and NF-{kappa}B (Nuclear Factor kappa-light-chain-enhancer of activated B cells) pathways, in the cellular response to GA-DM. While loss of genes involved in regulation of cholesterol biosynthesis conferred resistance to GA-DM, the disruption of ubiquitin-mediated proteolysis and the Hippo pathway genes sensitised cells to GA-DM. Inflammatory genes enriched at later time points suggests that a delayed cellular response contributes to cytotoxicity. Our findings propose a mechanistic model wherein GA-DM perturbs lipid and inflammatory pathways to exert cytotoxic effects and highlights potential targets to enhance its therapeutic efficacy. This work demonstrates the utility of functional genomics in elucidating natural product mechanisms and guiding rational drug development.

Published in Journal of Genetics · not in our set (fewer than 10 published preprints to learn from) · training set

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