Antiangiogenic Activity of 2-Deoxy-D-Glucose

Abstract

BackgroundDuring tumor angiogenesis, endothelial cells (ECs) are engaged in a number of energy consuming biological processes, such as proliferation, migration, and capillary formation. Since glucose uptake and metabolism are increased to meet this energy need, the effects of the glycolytic inhibitor 2-deoxy-D-glucose (2-DG) on in vitro and in vivo angiogenesis were investigated.Methodology/Principal FindingsIn cell culture, 2-DG inhibited EC growth, induced cytotoxicity, blocked migration, and inhibited actively forming but not established endothelial capillaries. Surprisingly, 2-DG was a better inhibitor of these EC properties than two more efficacious glycolytic inhibitors, 2-fluorodeoxy-D-glucose and oxamate. As an alternative to a glycolytic inhibitory mechanism, we considered 2-DG's ability to interfere with endothelial N-linked glycosylation. 2-DG's effects were reversed by mannose, an N-linked glycosylation precursor, and at relevant concentrations 2-DG also inhibited synthesis of the lipid linked oligosaccharide (LLO) N-glycosylation donor in a mannose-reversible manner. Inhibition of LLO synthesis activated the unfolded protein response (UPR), which resulted in induction of GADD153/CHOP and EC apoptosis (TUNEL assay). Thus, 2-DG's effects on ECs appeared primarily due to inhibition of LLOs synthesis, not glycolysis. 2-DG was then evaluated in two mouse models, inhibiting angiogenesis in both the matrigel plug assay and the LHBETATAG transgenic retinoblastoma model.Conclusions/SignificanceIn conclusion, 2-DG inhibits endothelial cell angiogenesis in vitro and in vivo, at concentrations below those affecting tumor cells directly, most likely by interfering with N-linked glycosylation rather than glycolysis. Our data underscore the importance of glucose metabolism on neovascularization, and demonstrate a novel approach for anti-angiogenic strategies.

Combination of glycolysis inhibition with chemotherapy results in an antitumor immune response

“…3 E and F), suggesting that the use of 2DG in vivo does not impair T-cell functions. Also, a recent study suggested that, under normoxic conditions, the primary activity of 2DG might not be to inhibit glycolysis but rather to interfere with N-linked glycosylation, leading to ER stress (4,28). It is of course important to keep in mind that escape from the immune-mediated surveillance of dying cells is among the characteristics of tumor cells.…”

A competitive hexapeptide inhibitor of annexin A2 prevents hypoxia-induced angiogenic events

“…The role of the N-terminal peptide in inhibiting angiogenesis was tested in the in vivo murine Matrigel plug assay, as previously described (Auerbach et al, 2003;Merchan et al, 2010). Mice were injected with Matrigel in the presence of PBS alone (negative control) or Matrigel mixed with VEGF and basic FGF (bFGF; positive control).…”

“…The Matrigel plug assay was performed as described previously (Merchan et al, 2010). Briefly, 500 l of unpolymerized Matrigel (BD Biosciences) (~20 mg/ml), either alone (negative control), mixed with bFGF and VEGF (100 ng/ml each), or in the presence of 5 M of the LGKLSL or LCKLSL peptides in combination with bFGF and VEGF (treatment group), was injected subcutaneously at the left lower abdominal wall of four groups of athymic nude mice (4-to 6-weeks-old) (Harlan, Madison, WI).…”

“…Briefly, 500 l of unpolymerized Matrigel (BD Biosciences) (~20 mg/ml), either alone (negative control), mixed with bFGF and VEGF (100 ng/ml each), or in the presence of 5 M of the LGKLSL or LCKLSL peptides in combination with bFGF and VEGF (treatment group), was injected subcutaneously at the left lower abdominal wall of four groups of athymic nude mice (4-to 6-weeks-old) (Harlan, Madison, WI). Mice were killed 5 days after the Matrigel injections, and the plugs were recovered, photographed, fixed in formaldehyde and embedded in OCT. Quantification of FITC-Dextran levels in the plugs was performed as described previously (Merchan et al, 2010;Klement et al, 2000).…”

2-deoxy-D-glucose combined with ferulic acid enhances radiation response in non-small cell lung carcinoma cells

“…In these cells, flux of glucose through the pentose phosphate shunt is required for efficient generation of NADPH, which maintains the high ratio of reduced to oxidized glutathione (GSH/GSSG) required for optimal control of oxidative stress; in addition, glycolytic flux generates pyruvate, which can function as a peroxide scavenger [9]. The increased glucose uptake in cancer cells is most probably a requirement to meet multiple processes including antioxidant defenses, protein glycosylation and angiogenesis [10,11]. It has been suggested that many cancers may be selectively susceptible to oxidative damage if systemic glucose deprivation is imposed.…”