Metabolomics identifies the intersection of phosphoethanolamine with menaquinone-triggered apoptosis in an in vitro model of leukemia

Abstract

Altered metabolism is increasingly acknowledged as an important aspect of cancer, and thus serves as a potentially fertile area for the identification of therapeutic targets or leads. Our recent work using transcriptional data to predict metabolite levels in cancer cells led to preliminary evidence of the antiproliferative role of menaquinone (Vitamin K2) in the Jurkat cell line model of acute lymphoblastic leukemia. However, nothing is known about the direct metabolic impacts of menaquinone in cancer, which could provide insights into its mechanism of action. Here, we used metabolomics to investigate the process by which menaquinone exerts antiproliferative activity on Jurkat cells. We first validated the dose-dependent, semi-selective, pro-apoptotic activity of menaquinone treatment on Jurkat cells relative to non-cancerous lymphoblasts. We then used mass spectrometry-based metabolomics to identify systems-scale changes in metabolic dynamics that are distinct from changes induced in non-cancerous cells or by other chemotherapeutics. One of the most significantly affected metabolites was phosphoethanolamine, which exhibited a two-fold increase in menaquinone-treated Jurkat cells compared to vehicle-treated cells at 24 h, growing to a five-fold increase at 72 h. Phosphoethanolamine elevation was observed prior to the induction of apoptosis, and was not observed in menaquinone-treated lymphoblasts or chemotherapeutic-treated Jurkat cells. We also validated the link between menaquinone and phosphoethanolamine in an ovarian cancer cell line, suggesting potentially broad applicability of their relationship. This metabolomics-based work is the first detailed characterization of the metabolic impacts of menaquinone treatment and the first identified link between phosphoethanolamine and menaquinone-induced apoptosis.

Preclinical Development of a Nontoxic Oral Formulation of Monoethanolamine, a Lipid Precursor, for Prostate Cancer Treatment

“…PhosE exhibits antitumor activity in various in vitro and in vivo models by affecting multiple signaling pathways (9–15). Interestingly, vitamin K2-induced apoptosis in Jurkat cells is ascribed to intracellular PhosE accumulation (18). …”

Manipulation of metabolism in complex eukaryotic systems to control cellular state

“…Other work showed that menaquinone can have therapeutic effects on prostate cancer cells, whether or not they were hormone-dependent, via caspase-3 and -8 dependent apoptosis [45]. Interestingly, recent metabolite profiling-driven work identified an unknown connection between menaquinone and phosphoethanolamine in the induction of apoptosis in a leukemic cell line [46]. This is particularly relevant given a recent series of papers showing the anti-cancer effects of phosphoethanolamine on breast cancer, leukemia, and renal cell carcinoma [47][48][49].…”

Potencial antitumoral da formulação lipossomal DODAC/fosfoetanolamina sintética no modelo de hepatocarcinoma

“…A capacidade da FO-S em promover apoptose, em diferenças linhagens de células tumorais, tem sido demostrada em diversos estudos publicados (Ferreira et al, 2011;Ferreira et al, 2012a;Ferreira et al, 2012b;Ferreira et al, 2013b;Ferreira et al, 2013c;Luna et al, 2014;Luna et al, 2016). Interessantemente, a apoptose desencadeada pela vitamina K2, em células de leucemia aguda humana Jurkat, foi atribuída ao acumulado intracelular de fosfoetanolamina (Dhakshinamoorthy et al, 2015).…”

“…A internalização do TRAIL e seus receptores pode recrutar fatores que induzem a permeabilização da membrana lisossomal, resultando na liberação da catepsina B para o citosol Guicciardi, 2014 A análise morfológica por MEV, das células tumorais B16F10 tratadas com os lipossomas DODAC/FO-S, evidenciou que as células morrem após a ativação de mecanismos pró-apoptóticos, conforme discutido anteriormente e relatado nas publicações com a FO-S (Ferreira et al, 2011;Ferreira et al, 2012a;Ferreira et al, 2012b;Ferreira et al, 2013b;Ferreira et al, 2013c;Luna et al, 2014;Dhakshinamoorthy et al, 2015;Luna et al, 2016). Diversos trabalhos descrevem as alterações que ocorrem nas células apoptóticas, como a formação de bolhas na membrana, seguida por cariorrexia, e a separação de fragmentos de células em corpos apoptóticos, durante um processo chamado "budding" (Elmore, 2007;Goldar et al, 2015).…”

“…Dessa forma, os resultados mostram que o efeito apoptótico da fosfoetanolamina apresenta uma importante função na indução de morte celular após o tratamento com vitamina K2 (Dhakshinamoorthy et al, 2015).…”