The Transcription Factor Myc Controls Metabolic Reprogramming upon T Lymphocyte Activation

“The metabolic changes associated with T cell activation are reminiscent of the characteristic metabolic activities of tumor cells and may represent a general metabolic reprogramming during cell growth and proliferation (Kim and Dang, 2006; Vander Heiden et al., 2009; Warburg, 1956).”

“The comparison of cytokine gene expression in resting and active T cells was visualized as heat maps with Cluster (Eisen and Brown, 1999) and TreeView (Eisen et al., 1998).”

“The metabolic changes associated with T cell activation are reminiscent of the characteristic metabolic activities of tumor cells and may represent a general metabolic reprogramming during cell growth and proliferation (Kim and Dang, 2006; Vander Heiden et al., 2009; Warburg, 1956).”

“Whereas prokaryotes and unicellular eukaryotes rely primarily on homeostatic regulation of cell metabolism, metabolic programs in multicellular eukaryotes have evolved to be actively controlled by extracellular signals (Deberardinis et al., 2008).”

“Metabolic flux analysis in a glioblastoma cell line further revealed that a major portion of glutamine-derived nitrogen and carbon are incorporated into amino acids and lactate, a fraction of which are secreted as metabolic by-products (DeBerardinis et al., 2007).”

“Recent studies in human cell lines have suggested that enforced expression of Myc regulates the rate-limiting glutaminolyic enzyme glutaminase 1 (Gls1) through both transcription and posttranscriptional mechanisms (Gao et al., 2009; Wise et al., 2008), and p53 regulates the transcription of glutaminase 2 (Gls2) (Hu et…”

“Animals Hif1a and p53-null mice on the C57BL/6 background were purchased from the Jackson Laboratory (Jacks et al., 1994; Ryan et al., 2000) and Hif1a mice were crossed with ROSA26CreERT2 (de Luca et al.”

“Because a-KG is required to maintain cell viability and minimal cell proliferation in the absence of glutamine (Wise et al., 2008), addition of a-KG alone was sufficient to drive only one cell division in the absence of glutamine.”

“To further test the dependency of T cell proliferation on glucose and glutamine catabolism in vivo, we applied 2DG and DON in an adoptive transfer model by using ovalbumin (OVA)specific, TCR-transgenic OT-II T cells (Figure 1G; Shi et al., 2011).”

“Because glutamine flux might regulate cell growth through the mTOR pathway (Nicklin et al., 2009), glutamine-dependent modulation of mTOR activation might partially account for the regulation of T cell growth by Myc.”

“, 2008), and the mammalian target of rapamycin (mTOR), an essential component of PI3K-Akt pathway, has recently been implicated in regulating fatty acid metabolism in memory T cells (Pearce et al., 2009; Araki et al., 2009).”

“, 2008), and the mammalian target of rapamycin (mTOR), an essential component of PI3K-Akt pathway, has recently been implicated in regulating fatty acid metabolism in memory T cells (Pearce et al., 2009; Araki et al., 2009).”

“Signaling via ERK and AKT pathways has been implicated in the uptake of glutamine and glucose, respectively (Carr et al., 2010; Frauwirth et al., 2002; Wofford et al., 2008), and the mammalian target of rapamycin (mTOR), an essential component of PI3K-Akt pathway, has recently been implicated in regulating fatty acid…”

“The metabolic changes associated with T cell activation are reminiscent of the characteristic metabolic activities of tumor cells and may represent a general metabolic reprogramming during cell growth and proliferation (Kim and Dang, 2006; Vander Heiden et al., 2009; Warburg, 1956).”

“Consistent with a direct role ofMyc in regulating cell proliferation (Dang et al., 2006; Eilers and Eisenman, 2008), addition of a-KG with HT and PAs failed to promote cell proliferation in Myc-deficient T cells (Figure S7C).”

“Notably, ectopic overexpression of Myc not only alters the transcription of several glycolytic genes but also preferentially switches the expression of PKM1 to PKM2 by modulating the alternative splicing of the PKM transcript (Dang and Semenza, 1999; David et al., 2010).”

“T cell activation-induced glycolysis and glutaminolysis is reminiscent of the metabolic changes in tumor cells, where both aerobic glycolysis (the Warburg effect) and glutaminoly- sis are driven by aberrant oncogenic signals (Dang and Semenza, 1999; Vander Heiden et al., 2009).”

“The comparison of cytokine gene expression in resting and active T cells was visualized as heat maps with Cluster (Eisen and Brown, 1999) and TreeView (Eisen et al.”

“Whereas prokaryotes and unicellular eukaryotes rely primarily on homeostatic regulation of cell metabolism, metabolic programs in multicellular eukaryotes have evolved to be actively controlled by extracellular signals (Deberardinis et al., 2008).”

“Consistent with a direct role ofMyc in regulating cell proliferation (Dang et al., 2006; Eilers and Eisenman, 2008), addition of a-KG with HT and PAs failed to promote cell proliferation in Myc-deficient T cells (Figure S7C).”

“Primer sequences were obtained from PrimerBank (Spandidos et al., 2010) except for Gls1 and Gls2, which were manually designed.”

“Although recent studies indicate that the tumor suppressor p53 is required for the induction of GLS2 in a human tumor cell line (Hu et al., 2010; Suzuki et al., 2010),”

“We observed that acute depletion of Myc, in vivo, did not compromise the health of the animals over a period of 1 week, consistent with a recent report (Soucek et al., 2008).”

“TCR- and costimulator-mediated signals drive T cell growth and clonal expansion, and coordinately reprogram metabolic cascades to fulfill the ensuing bioenergetic and biosynthetic demands (Fox et al., 2005; Krauss et al., 2001).”

“Early studies have implicated the involvement of ERK-, AKT-, and mTOR-mediated signaling pathways in regulating T cell metabolism (Carr et al., 2010; Frauwirth et al., 2002; Pearce et al., 2009).”

“…and Myc were recently suggested to regulate 874 Immunity 35, 871–882, December 23, 2011 ª2011 Elsevier Inc. cell metabolism in transformed cells (Dang and Semenza, 1999; Gordan et al., 2007) and were highly induced at both the transcription and protein levels upon T cell activation (Figures 2D–2G).”

“Although recent studies indicate that the tumor suppressor p53 is required for the induction of GLS2 in a human tumor cell line (Hu et al., 2010; Suzuki et al., 2010),”

“Arg1flox/flox; Tie2Cre mice that were backcrossed to the C57BL/6 background were obtained from P. Murray (SJCRH, Memphis) (El Kasmi et al., 2008).”

“Consequently, one of the fundamental problems in animal cells is coordination of the metabolic program with cell growth- and proliferation-associated bioenergetic demand (Jorgensen and Tyers, 2004).”

“Early studies showed that polyclonal mitogens such as concanavalin A and phytohaemagglutinin drive the upregulation of glycolysis and glutamine oxidation in thymocytes and mature lymphocytes (Brand et al., 1984; Newsholme et al., 1985b).”

“Although Myc was suggested to be required for T cell proliferation and growth under most experimental conditions (Dose et al., 2006; Iritani et al., 2002), one study indicated that it is essential for cell proliferation but not cell growth (Trumpp et al., 2001).”

“…via ERK and AKT pathways has been implicated in the uptake of glutamine and glucose, respectively (Carr et al., 2010; Frauwirth et al., 2002; Wofford et al., 2008), and the mammalian target of rapamycin (mTOR), an essential component of PI3K-Akt pathway, has recently been implicated in…”

“During proliferation, T cells can undergo a cell division every 4–6 hr (van Stipdonk et al., 2003).”

“Mycflox/flox mice were obtained from F. Alt (Harvard) (de Alboran et al., 2001) and were backcrossed five generations onto the C56BL/6 background before breeding to ROSA26CreER(Tam) (Badea et al., 2003).”

“We generated a mouse model carrying a conditional HIF1a allele (Hif1aflox/flox) and a tamoxifen-inducible Cre recombinase (CreERT2) transgene (de Luca et al., 2005; Ryan et al., 2000) to delete HIF1a flox alleles in an acute manner.”

“Glutamine has long been known to be an indispensible nutrient for proliferating cells (Kovacevic and McGivan, 1983).”

“Early studies showed that polyclonal mitogens such as concanavalin A and phytohaemagglutinin drive the upregulation of glycolysis and glutamine oxidation in thymocytes and mature lymphocytes (Brand et al., 1984; Newsholme et al., 1985b).”

“Freshly isolated murine primary T cells were either maintained in interleukin 7 (IL-7) (hereafter ‘‘resting T cells’’) (Vella et al., 1997) or were stimulated with anti-CD3 plus anti-CD28 (hereafter ‘‘active T cells’’).”

“, 2001) and were backcrossed five generations onto the C56BL/6 background before breeding to ROSA26CreER(Tam) (Badea et al., 2003).”

“Antigen activation of T lymphocytes drives distinct transcriptional programs including upregulation of cell cycle and cytokine genes (Lu et al., 2004; Teague et al., 1999).”

“Intriguingly, the metabolic enzymes Aldh18A1, Prodh, and OAT that link glutamine and proline to ornithine synthesis were induced in a Mycdependent manner upon T cell activation (Figures 7C and S5F), as were the metabolic enzymes, ODC, spermine synthase (SRM), and spermidine synthase (SMS), that convert ornithine to po…”

“I Myc, but Not HIF1a, Is Required for Activation-Induced T Cell Metabolic Reprogramming With an in silico approach (Roider et al., 2009), transcription factors possibly involved in activation-induced T cell metabolic mmunity 35, 871–882, December 23, 2011 ª2011 Elsevier Inc. 873 (A andB) The…”

“TCR- and costimulator-mediated signals drive T cell growth and clonal expansion, and coordinately reprogram metabolic cascades to fulfill the ensuing bioenergetic and biosynthetic demands (Fox et al., 2005; Krauss et al., 2001).”

“…studies clearly demonstrate that Myc directly regulates T cell proliferation mainly through transcriptional control of cell cycle regulators (Dose et al., 2006; Iritani et al., 2002), our studies implicate Myc as an essential coordinator of T cell activation-induced cell growth,…”

“We compared our metabolic gene expression data of active mouse T lymphocytes with metabolic gene expression data from a recently published microarray study of active human T lymphocytes (Wang et al., 2008).”

“…demonstrate that Myc directly regulates T cell proliferation mainly through transcriptional control of cell cycle regulators (Dose et al., 2006; Iritani et al., 2002), our studies implicate Myc as an essential coordinator of T cell activation-induced cell growth, proliferation, and several…”

“…action of arginase as part of the urea cycle (Morris, 2004), urea cycle enzymes including arginase were not induced upon activation (Figure S7D) nor were any defects in proliferation observed in T cells lacking arginase I (Figures 7A and 7B), the dominant isoform in the thymus (Yu et al., 2003).”

“Early studies showed that polyclonal mitogens such as concanavalin A and phytohaemagglutinin drive the upregulation of glycolysis and glutamine oxidation in thymocytes and mature lymphocytes (Brand et al., 1984; Newsholme et al., 1985b).”

“Previous studies in bovine endothelial and intestinal epithelial cells suggested the possible presence of a noncanonical metabolic pathway, which generates ornithine from glutamine (Wu et al., 2000; Wu and Morris, 1998).”

“Coinciding with the clearance of antigen, effector T cells undergo an abrupt contraction phase in which the majority of T cells are eliminated by apoptosis (Green, 2003; Schumacher et al., 2010).”

“Antigen activation of T lymphocytes drives distinct transcriptional programs including upregulation of cell cycle and cytokine genes (Lu et al., 2004; Teague et al., 1999).”

“Consistent with a previous study (Bowlin et al., 1987), difluoromethylornithine (DFMO), a potent inhibitor of ornithine decarboxylase (ODC) and polyamine biosynthesis, inhibited activation-induced T cell proliferation in vitro and in vivo (Figure 6F and 1H), and the addition of exogenous polyamines completely restored…”

“Consistent with previous work (Rowell and Wells, 2006), T cell activation was associated with downregulation of the cell cycle inhibitor p27, the upregulation of cyclins and cyclin-dependent kinases (CDKs), and the appearance of markers of S phase, such as the phosphorylation of RB and Cdc6 (Figure…”

“Coinciding with the clearance of antigen, effector T cells undergo an abrupt contraction phase in which the majority of T cells are eliminated by apoptosis (Green, 2003; Schumacher et al., 2010).”

“We assessed the protein expression of glutamate dehydrogenase (Glud1), which catalyzes the oxidative deamination of glutamate to a-ketoglutarate, an important process for energy homeostasis in T cells (Wang et al., 2011).”

“Loss of Myc was associated with profoundly diminished expression of genes involved in metabolism and the inability for normal primary T cells to grow and proliferate (Wang et al., 2011a).”