Ascorbate Plus Buformin in AML: A Metabolic Targeted Treatment

Abstract

In the present study, we characterized the metabolic background of different Acute Myeloid Leukemias’ (AMLs) cells and described a heterogeneous and highly flexible energetic metabolism. Using the Seahorse XF Agilent, we compared the metabolism of normal hematopoietic progenitors with that of primary AML blasts and five different AML cell lines. We assessed the efficacy and mechanism of action of the association of high doses of ascorbate, a powerful oxidant, with the metabolic inhibitor buformin, which inhibits mitochondrial complex I and completely shuts down mitochondrial contributions in ATP production. Primary blasts from seventeen AML patients, assayed for annexin V and live/dead exclusion by flow cytometry, showed an increase in the apoptotic effect using the drug combination, as compared with ascorbate alone. We show that ascorbate inhibits glycolysis through interfering with HK1/2 and GLUT1 functions in hematopoietic cells. Ascorbate combined with buformin decreases mitochondrial respiration and ATP production and downregulates glycolysis, enhancing the apoptotic effect of ascorbate in primary blasts from AMLs and sparing normal CD34+ bone marrow progenitors. In conclusion, our data have therapeutic implications especially in fragile patients since both agents have an excellent safety profile, and the data also support the clinical evaluation of ascorbate–buformin in association with different mechanism drugs for the treatment of refractory/relapsing AML patients with no other therapeutic options.

Choosing the Right Cell Line for Acute Myeloid Leukemia (AML) Research

“…They were used, among others, to determine the effect of ascorbate and buformin on the metabolic activity of AML cells. Studies confirmed the clinical assessment of the efficacy of the combination of these compounds in treating refractory cases of AML [ 99 ]. Bioenergetic analysis on OCI-AML2 and OCI-AML3 lines showed that AMPK / PERK activation by GSK621 induces mitochondrial apoptosis in leukemic cells [ 100 ].…”

The Anti-Leukemia Effect of Ascorbic Acid: From the Pro-Oxidant Potential to the Epigenetic Role in Acute Myeloid Leukemia

“…Moreover, ascorbate also shows the ability to induce cell death by targeting glycolytic metabolism in primary AML blasts, through the inhibition of hexokinase 1/2 (HK1/2) and GLUT1 in hematopoietic cells, and, in combination with the metabolic inhibitor buformin, also decreases mitochondrial respiration and ATP production, sparing healthy CD34 + cells. Overall, these data clearly depict an effect of ascorbate on glycolysis and contribute to elucidate the targets and mechanisms through which this therapeutic agent exerts its anti-cancer effects (Banella et al, 2022).…”

Effects of Triheptanoin on Mitochondrial Respiration and Glycolysis in Cultured Fibroblasts from Neutral Lipid Storage Disease Type M (NLSD-M) Patients

“…The goal of this study was to evaluate if triheptanoin can ameliorate the glycolytic and oxidative metabolism when administered to fibroblasts of patients with NLSD-M. The method used to analyze the oxidative metabolism of affected fibroblasts has been widely used in other studies, mainly for oncological pathologies [ 7 , 8 , 10 ], but has never been used for the study of NLSD. The results demonstrate that triheptanoin ameliorates the metabolic performances in NLSD-M fibroblasts.…”

“…To achieve this, cells use the spare respiratory capacity (SRC). This process is tightly controlled by the nature and flow of nutrients that can be oxidized in the mitochondrial matrix by the tricarboxylic acid (TCA) cycle [ 8 ]. SRC is increased in all NLSD-M patients after treatment with triheptanoin, indicating that acetyl-CoA and Propionyl-CoA, the metabolized products of triheptanoin, provide an anaplerotic effect by replenishing deficient TCA cycle intermediates in NLSD-M. SRC represents a particularly robust functional parameter to evaluate mitochondrial reserve.…”

“…Vockley et al demonstrated that triheptanoin treatment is associated with reduced major clinical events, such as rhabdomyolysis, cardiomyopathy, and hypoglycemia in patients with long-chain fatty acid oxidation disorders [ 7 ]. However, in studies on mice, using long-term supplementation of triheptanoin induces de novo biosynthesis and elongation of fatty acids in both WT and VLCAD-/- mice, altering the hepatic and cardiac fatty acid composition to no physiological profiles, characterized by a strong reduction in polyunsaturated fatty acids and a marked increase in monounsaturated species [ 8 ]. These results suggest that the time of application and the dose should be individually adapted to meet the energy demand of the patient.…”