O-cyclic phytosphingosine-1-phosphate stimulates HIF1α-dependent glycolytic reprogramming to enhance the therapeutic potential of mesenchymal stem cells

Abstract

O-cyclic phytosphingosine-1-phosphate (cP1P) is a novel chemically synthesized sphingosine metabolite derived from phytosphingosine-1-phosphate. Although structurally similar to sphingosine-1-phosphate (S1P), its biological properties in stem cells remain to be reported. We investigated the effect of cP1P on the therapeutic potential of mesenchymal stem cells (MSCs) and their regulatory mechanism. We found that, under hypoxia, cP1P suppressed MSC mitochondrial dysfunction and apoptosis. Metabolic data revealed that cP1P stimulated glycolysis via the upregulation of glycolysis-related genes. cP1P-induced hypoxia-inducible factor 1 alpha (HIF1α) plays a key role for MSC glycolytic reprogramming and transplantation efficacy. The intracellular calcium-dependent PKCα/mammalian target of the rapamycin (mTOR) signaling pathway triggered by cP1P regulated HIF1α translation via S6K1, which is critical for HIF1 activation. Furthermore, the cP1P-activated mTOR pathway induced bicaudal D homolog 1 expression, leading to HIF1α nuclear translocation. In conclusion, cP1P enhances the therapeutic potential of MSC through mTOR-dependent HIF1α translation and nuclear translocation.

Neuronal Metabolism and Neuroprotection: Neuroprotective Effect of Fingolimod on Menadione-Induced Mitochondrial Damage

“…All the effects of FP on glycolysis were mediated through its interaction with S1P1 receptors, as all of these effects disappeared when the S1P1 antagonist was present, being affected the basal glycolysis, glycolytic capacity, glycolytic reserve, and the capacity to maintain the aldolase activity. This finding was common with other authors who also found the need of interaction of FP with its receptor to develop its glycolytic protective actions [ 54 , 60 ]. In the study of G-6-PDH activity, the effect of FP was also mediated by the interaction with its specific S1P1 receptor, as the increase in G-6-PDH activity was totally abolished when the S1P1 antagonist was present in the incubation media.…”

“…When we studied the glycolytic reserve, which is the ability of cells to adapt to extra energetic demands when the glycolytic function is working at maximum, neurons incubated in presence of VitK3 also showed a reduction in glycolytic reserve by 48% compared to the control; again, coincubation in the presence of FP reverted this value to control levels (Figure 2a,d). Our results partially agree with those from Lee et al [54], who found an increase of glycolytic capacity with S1P mimetics in mesenchymal cells, although the authors did not find differences in glycolytic reserve, possibly due to the different type of cells. This recovery in glycolytic capacity would rapidly supply ATP in situations where it is needed and when the mitochondrial metabolism, although generally having a high capacity to produce ATP, is not totally functional, as demonstrated previously.…”

“…In this work, we found a decrease (23%) in aldolase activity after treatment of neurons with VitK3. Again, FP restored values to those found in control neurons (Figure 3), agreeing with Lee et al, who also found an increase in aldolase activity with S1P mimetics [54]; moreover, Geffin et al found an upregulation of genes involved in glycolysis, including aldolase, in cells exposed to HIV and treated with FP [60]. On the basis of these findings, FP could be considered as a new therapeutic approach to the treatment of neurodegenerative diseases showing a decrease in glycolysis, such as PD, AD, or MS [26].…”

“…In presence of VitK3, we found a great decrease (46% compared to control) in glycolysis, measured as ECAR after 2 h of incubation; coincubation in the presence of FP recovered values to control levels (Figure 2a,b). This effect was similar to that found with some sphingosine-1-phospate mimetics that increase the expression of genes related with glycolysis, ECAR, and glycolytic capacity [54]. In the study of glycolytic capacity, which measures the maximum rate of conversion of glucose to pyruvate or lactate, we found that VitK3 produced a decrease by 47% compared to the control, with this decrease being totally reverted by coincubation in the presence of FP (Figure 2a,c).…”

Mechanisms Underlying the Therapeutic Potential of Mesenchymal Stem Cells in Atherosclerosis

“…Zhang et al investigated the use of MSCs in myocardial infarction and showed that pre-treating MSCs with asprosin, an adipokine that inhibits reactive oxygen species (ROS) generation, significantly improved MSC homing and proliferation while reducing apoptosis and scar size [98]. Lee et al also demonstrated the effectiveness of pre-treatment with O-cyclic-phytosphingosine-1-phosphate which, through reversal of ROS damage, upregulated proliferation, migration and anti-apoptosis [99]. These studies propose innovative ways of rejuvenating autologous MSCs for the treatment of atherosclerosis.…”