Decoupling NAD+ metabolic dependency in chondrosarcoma by targeting the SIRT1-HIF-2α axis.

Chondrosarcomas represent the second most common primary bone malignancy. Despite the vulnerability of chondrosarcoma cells to nicotinamide adenine dinucleotide (NAD+) depletion, targeting the NAD+ synthesis pathway remains challenging due to broad implications in biological processes. Here, we establish SIRT1 as a central mediator reinforcing the dependency of chondrosarcoma cells on NAD+ metabolism via HIF-2α-mediated transcriptional reprogramming. SIRT1 knockdown abolishes aggressive phenotypes of chondrosarcomas in orthotopically transplanted tumors in mice. Chondrosarcoma cells thrive under glucose starvation by accumulating NAD+ and subsequently activating the SIRT1-HIF-2α axis. Decoupling this link via SIRT1 inhibition unleashes apoptosis and suppresses tumor progression in conjunction with chemotherapy. Unsupervised clustering analysis identifies a high-risk chondrosarcoma patient subgroup characterized by the upregulation of NAD+ biosynthesis genes. Finally, SIRT1 inhibition abolishes HIF-2α transcriptional activity and sensitizes chondrosarcoma cells to doxorubicin-induced cytotoxicity, irrespective of underlying pathways to accumulate intracellular NAD+. We provide system-level guidelines to develop therapeutic strategies for chondrosarcomas.

Effect of biotic elicitors on the accumulation of bilobalide and ginkgolides in Ginkgo biloba cell cultures.

The effect of biotic elicitors on the production of bilobalide and ginkgolides in Ginkgo biloba cell suspension cultures was studied. The treatment of cell cultures with Candida albicans and Staphylococcus aureus as elicitors increased the amounts of bilobalide (BB), ginkgolide A (GA) and ginkgolide B (GB), with slight growth inhibition. The native bacterial elicitor was more effective for secondary metabolite accumulations both in cells and culture medium than autoclaved. However, exposure times of the cells to the elicitors strongly influenced the production of BB, GA and GB. This study suggests that biotic elicitors can regulate the production of BB, GA and GB either directly or indirectly. These results also describe the establishment of optimum conditions that determine the effects of biotic elicitors on secondary metabolism of bilobalides.

Effect of supplementing terpenoid biosynthetic precursors on the accumulation of bilobalide and ginkgolides in Ginkgo biloba cell cultures.

The effect of precursor feeding on the production of bilobalide and ginkgolides was studied with suspension cell cultures of Ginkgo biloba. The precursors greatly influenced the productivity of bilobalide and ginkgolides. Precursor supplementation increased the accumulation of both bilobalide and ginkgolides, and with positive effect on cell growth. The GA accumulation by cell cultures was influenced by precursors upstream in the metabolism, whereas the BB accumulation was under the influence of downstream precursors of the terpenoid biosynthetic pathway. Furthermore, precursor feeding modified the ratios of the BB, GA and GB in cells and cell cultures of G. biloba. The studies also aid in understanding effect of precursor feeding on the bilobalide and ginkgolides biosynthetic pathway.

Biotransformation and impact of ferulic acid on phenylpropanoid and capsaicin levels in Capsicum annuum L. cv. P1482 cell suspension cultures.

Cell suspension cultures of Capsicum annuum L. cv. P1482 were fed with exogenous ferulic acid to monitor their biotransformation abilities. A portion of the ferulic acid was biotransformed into vanillin, a major natural flavor, and capsaicin, a principle secondary metabolite characteristic of Capsicum species. The cellular vanillin concentrations were relatively higher than capsaicin levels and were maximal (2 mg/g DW) 4 days after 0.6 mM ferulic acid feeding. Maximal vanillin levels in the culture medium were 10 mg/L at 4 and 3 days after feeding with 1.25 and 2.5 mM ferulic acid, respectively. With regard to capsaicin levels, the cellular levels were slightly decreased by ferulic acid feeding, whereas the levels in the culture medium were increased. Ferulic acid feeding not only enhanced vanillin and capsaicin production but also increased the concentrations of other phenylpropanoid metabolites.