Olanzapine and aripiprazole differentially affect glucose uptake and energy metabolism in human mononuclear blood cells

https://doi.org/10.1016/j.jpsychires.2016.12.012Get rights and content

Abstract

The use of antipsychotics carries the risk of metabolic side effects, such as weight gain and new onset type-2 diabetes mellitus. The mechanisms of the observed metabolic alterations are not fully understood. We compared the effects of two atypical antipsychotics, one known to favor weight gain (olanzapine), the other not (aripiprazole), on glucose metabolism. Primary human peripheral blood mononuclear cells (PBMC) were isolated and stimulated with olanzapine or aripiprazole for 72 h. Cellular glucose uptake was analyzed in vitro by 18F-FDG uptake. Further measurements comprised mRNA expression of glucose transporter (GLUT) 1 and 3, GLUT1 protein expression, DNA methylation of GLUT1 promoter region, and proteins involved in downstream glucometabolic processes. We observed a 2-fold increase in glucose uptake after stimulation with aripiprazole. In contrast, olanzapine stimulation decreased glucose uptake by 40%, accompanied by downregulation of the cellular energy sensor AMP activated protein kinase (AMPK). GLUT1 protein expression increased, GLUT1 mRNA expression decreased, and GLUT1 promoter was hypermethylated with both antipsychotics. Pyruvat-dehydrogenase (PDH) complex activity decreased with olanzapine only.

Our findings suggest that the atypical antipsychotics olanzapine and aripiprazole differentially affect energy metabolism in PBMC. The observed decrease in glucose uptake in olanzapine stimulated PBMC, accompanied by decreased PDH point to a worsening in cellular energy metabolism not compensated by AMKP upregulation. In contrast, aripiprazole stimulation lead to increased glucose uptake, while not affecting PDH complex expression. The observed differences may be involved in the different metabolic profiles observed in aripiprazole and olanzapine treated patients.

Introduction

According to current guidelines, antipsychotics are first line drugs for the treatment of schizophrenia (Baandrup et al., 2016, Lehman et al., 2004). Additionally, antipsychotics are increasingly used for many other psychiatric disorders such as bipolar mania and psychotic depression, in particular second generation antipsychotics (SGA) (Perlis et al., 2006, Tarr et al., 2011, Zhou et al., 2015). In adolescents and adults, there is a clear link between the use of antipsychotic treatment and weight gain, impaired glucose tolerance, and risk of developing type-2 diabetes mellitus, which differs across drugs (Farwell et al., 2004, Galling et al., 2016, Gianfrancesco et al., 2002, Lambert et al., 2006).

Studies on the effects of antipsychotics on cellular glucose transport yielded inconsistent results, depending on the model used. Ardizzone and colleagues demonstrated decreased glucose transport in rat pheochromocytoma cells (PC12) after incubation with risperidone, clozapine, and loxapine, and a decrease of glucose transport in a rat muscle cell line (L6) with clozapine and fluphenazine, thereby suggesting that these drugs may inhibit glucose transport directly at the glucose transporter (GLUT) (Ardizzone et al., 2001). Dwyer and colleagues observed a decrease of glucose uptake after incubation with fluphenazine, chlorpromazine, clozapine and haloperidol, followed by an increase of GLUT1 and GLUT3 expression on the protein level (Dwyer et al., 1999). In contrast, in a model of chronic antipsychotic treatment, no differences in GLUT1 and GLUT3 mRNA expression were found in rat adipocytes, using haloperidol, olanzapine and ziprasidone as antipsychotic drugs (Minet-Ringuet et al., 2007). In a recent work by Babkin and colleagues, olanzapine induced inhibition of glucose transport was associated with specific binding in a polar region of the cytosolic part of the GLUT1 in a Staphylococcus epidermidis model glucose transporter, thereby inhibiting glucose transport (Babkin et al., 2015). Few studies examined the effects of antipsychotics in human cells (U937). Heiser and colleagues found an increase of mRNA expression for GLUT4 and GLUT5 after 48 h incubation with haloperidol and olanzapine, but no effect on GLUT1-3 (Heiser et al., 2006) in human leukemic blood cells. Taken together these results suggest that antipsychotics may affect glucose transport under particular conditions.

Studies so far were performed in rat or human tumor cell lines, and in an E. coli model of glucose transporter (Ardizzone et al., 2001, Babkin et al., 2015, Heiser et al., 2006). We are not aware of studies using primary human cell cultures. We here used primary cell cultures of peripheral blood mononuclear cells (PBMC), because PBMC express the glucose transporter (GLUT) 1 and 3 (Daneman et al., 1992, Maratou et al., 2007), and various neurotransmitter receptors such as dopaminergic receptors (Leite et al., 2016), serotonergic receptors (Ahangari et al., 2013, Mizrahi et al., 2004) and acetylcholine receptors, which are similar to those found in the brain (Neumann et al., 2007, Tayebati and Amenta, 2008). This makes PBMC a potentially useful model for investigating mechanisms involved in peripheral and central metabolic abnormalities during antipsychotic drug action.

We here examined glucose uptake, GLUT1 (SLC2A1) and GLUT3 (SLC2A3) expression and DNA methylation of the GLUT1 promoter region in human PBMC obtained from healthy blood donors. Additionally, relevant downstream factors of glucose metabolism were examined, i.e. the expression of the pyruvate dehydrogenase (PDH) complex, expression and phosphorylation of the AMP-activated protein kinase (AMPK), a key regulator of cellular energy balance (Hardie, 2008), and the protein expression level of glycerinaldehy-3-phophate dehydrogenase (GADPH). We used two antipsychotics with different side effect profiles for PBMC stimulation, one known to be associated with metabolic side effects including weight gain and disturbance of glucose metabolism (olanzapine) and one with less effects on metabolic functions (aripiprazole) (Correll et al., 2015, Newcomer, 2005). We hypothesized that olanzapine decreases cellular energy metabolism compared to aripiprazole.

Section snippets

Isolation and cultivation of human peripheral mononuclear cells

The study was approved by the local ethics committee. All samples were obtained anonymized from the blood preservation unit of the Hannover Medical School.

Peripheral mononuclear cells (PBMC) from healthy blood donors were isolated from leucocyte filters obtained from the blood transfusion service at Hannover Medical School by use of ficoll gradient as follows. Cells were separated from filters by rinsing with PBS (Biochrom AG, #15140-122), 35 mL of the blood/PBS mixture were loaded on top of

Olanzapine and aripiprazole differentially regulate cellular glucose uptake in vitro in a dose-dependent manner

As olanzapine and aripiprazole distinctly influence whole body glucose metabolism we assessed whether prolonged stimulation with these compounds impacts the metabolic status of PBMC in vitro. To assess whether olanzapine and aripiprazole directly affect cellular substrate uptake, PBMC were stimulated with indicated concentrations of olanzapine or aripiprazole for 72 h and cellular glucose uptake was analyzed by in vitro 18F-FDG uptake. Chronic stimulation with olanzapine resulted in a

Discussion

The main finding of our study is a differential regulation of glucose uptake by olanzapine and aripiprazole, pointing to decreased glucose uptake in PBMC by olanzapine, accompanied by decreased expression of the pyruvate dehydrogenase (PDH) complex, and decreased expression and activity of AMPK. In contrast, stimulation of PBMC with aripiprazole increased glucose uptake without altering PDH complex expression, and this was accompanied by AMPK deactivation. On the protein level, stimulation with

Role of the funding source

No source of funding.

Authors' contributions

All authors made substantive contribution to the study and qualified for authorship. In particular, all authors made 1) substantial contributions to the conception and design, or acquisition of data, or analysis and interpretation of the data, 2) were involved in drafting the manuscript, and 3) gave final approval of the version to be published.

Conflicts of interests

Kai G. Kahl has received speaker honoraria and a research grant by Servier, and speaker honoraria by Lundbeck, GSK, AstraZeneca, EliLilly and Otsuka.

Helge Frieling received speaker honoriaria by Astra Zeneca, Lilly, Janssen, Servier, Otsuka and Allergan, and a research grant from Bayer vital.

Stefan Bleich received speaker honoraria from Servier and Lundbeck.

Britta Stapel, Denise Hilfiker-Kleiner and Alexandra Kotsiari reported no financial disclosure.

Acknowledgments

The authors wish to thank Vanessa Buchholz and Patricia Fiedler for technical assistance.

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