Tumor cell metabolism was the discovery that the tumorassociated shift to aerobic glycolysis was controlled by the balance of expression of 2 isoforms of the glycolytic eFT508 enzyme pyruvate kinase M (PKM)[6]. Pyruvate kinase converts phosphoenolpyruvate (PEP) to pyruvate and is a SM5688 web rate-limiting enzyme in glycolysis [7]. 
PKM is one of three PKs (PKM, PKL, which is expressed only in the liver, and PKR which is expressed only in erythrocytes)[8] but the only form associated with cancer. The two isoforms of PKM are encoded by a single gene on chromosome 15, and are derived from a single transcript that gives rise to PKM1 mRNA and PKM2 mRNA by differential splicing [9]. The processed PKM1 mRNA excludes exon 10 while the processed PKM2 mRNA excludes exon 9, making the two transcripts best distinguishable by quantitative PCR. The PKM1 protein isPyruvate Kinase Modulation in Brain Tumorsconstitutively active, has a high affinity for its substrate PEP, converts PEP to pyruvate, and through complexation with other enzymes of the glycolytic pathway assures that pyruvate is moved into the mitochondria for use in the citric acid cycle [10]. Although PKM1 is frequently stated to be expressed in most adult normal tissues, the most stringent analyses suggest that it is expressed only in tissues in which a large amount of energy is produced such as muscle and brain [11]. In these tissues PKM1 expression is 
associated with a high PK activity and the oxidative phosphorylation that helps generate maximal amounts of ATP. The PKM2 protein in contrast is expressed in some differentiated tissues such as lung, but primarily is expressed in embryonic tissues and tumor cells [8,12,13]. PKM2 can exist in active tetrameric and nearly inactive dimeric forms although in tumors it is reported to exist nearly exclusively as a dimer [14]. 23977191 The dimeric form of PKM2 has a low affinity for PEP, is not associated with enzymes of the glycolytic complex, and favors conversion of pyruvate to lactate [7,15]. As a result of the inability of the PKM2 dimer to stimulate pyruvate production, glycolysis slows and the intermediates of glucose metabolism back-up in tumor cells. These intermediates in turn are used to generate the nucleotides and amino acids that are required for cell proliferation, increased biomass, and tumor growth [16,17]. Tumor formation therefore appears to involve alterations in PKM isoform expression, accompanied by a tumorpromoting shift in PK activity and metabolism from oxidative phosphorylation to aerobic glycolysis [6,18]. Although 24786787 tumors in general are thought to exhibit high levels of PKM2 and low levels of PK activity, this belief is based primarily on PKM2 RNA, PKM2 protein, or PK activity analysis (but rarely all three) of a limited number of high grade tumors. As a result, there is little information as to how changes in PKM mRNA levels relate to changes in PKM protein levels and PK activity, whether PKM isoform expression and activity changes along a continuum of grades tumors of the same histologic subtype, and what the consequences of these changes might be for tumor growth. To address these questions we examined PKM isoform expression and PK activity in an extensive series of grade I-IV astrocytomas, and have examined the consequences of alteration of PKM expression and activity on the growth of human glioma cells. The results of the studies show that while PK activity is uniformly low in all glioma, PKM2 expression is disproportionately up-regulated in.Tumor cell metabolism was the discovery that the tumorassociated shift to aerobic glycolysis was controlled by the balance of expression of 2 isoforms of the glycolytic enzyme pyruvate kinase M (PKM)[6]. Pyruvate kinase converts phosphoenolpyruvate (PEP) to pyruvate and is a rate-limiting enzyme in glycolysis [7]. PKM is one of three PKs (PKM, PKL, which is expressed only in the liver, and PKR which is expressed only in erythrocytes)[8] but the only form associated with cancer. The two isoforms of PKM are encoded by a single gene on chromosome 15, and are derived from a single transcript that gives rise to PKM1 mRNA and PKM2 mRNA by differential splicing [9]. The processed PKM1 mRNA excludes exon 10 while the processed PKM2 mRNA excludes exon 9, making the two transcripts best distinguishable by quantitative PCR. The PKM1 protein isPyruvate Kinase Modulation in Brain Tumorsconstitutively active, has a high affinity for its substrate PEP, converts PEP to pyruvate, and through complexation with other enzymes of the glycolytic pathway assures that pyruvate is moved into the mitochondria for use in the citric acid cycle [10]. Although PKM1 is frequently stated to be expressed in most adult normal tissues, the most stringent analyses suggest that it is expressed only in tissues in which a large amount of energy is produced such as muscle and brain [11]. In these tissues PKM1 expression is associated with a high PK activity and the oxidative phosphorylation that helps generate maximal amounts of ATP. The PKM2 protein in contrast is expressed in some differentiated tissues such as lung, but primarily is expressed in embryonic tissues and tumor cells [8,12,13]. PKM2 can exist in active tetrameric and nearly inactive dimeric forms although in tumors it is reported to exist nearly exclusively as a dimer [14]. 23977191 The dimeric form of PKM2 has a low affinity for PEP, is not associated with enzymes of the glycolytic complex, and favors conversion of pyruvate to lactate [7,15]. As a result of the inability of the PKM2 dimer to stimulate pyruvate production, glycolysis slows and the intermediates of glucose metabolism back-up in tumor cells. These intermediates in turn are used to generate the nucleotides and amino acids that are required for cell proliferation, increased biomass, and tumor growth [16,17]. Tumor formation therefore appears to involve alterations in PKM isoform expression, accompanied by a tumorpromoting shift in PK activity and metabolism from oxidative phosphorylation to aerobic glycolysis [6,18]. Although 24786787 tumors in general are thought to exhibit high levels of PKM2 and low levels of PK activity, this belief is based primarily on PKM2 RNA, PKM2 protein, or PK activity analysis (but rarely all three) of a limited number of high grade tumors. As a result, there is little information as to how changes in PKM mRNA levels relate to changes in PKM protein levels and PK activity, whether PKM isoform expression and activity changes along a continuum of grades tumors of the same histologic subtype, and what the consequences of these changes might be for tumor growth. To address these questions we examined PKM isoform expression and PK activity in an extensive series of grade I-IV astrocytomas, and have examined the consequences of alteration of PKM expression and activity on the growth of human glioma cells. The results of the studies show that while PK activity is uniformly low in all glioma, PKM2 expression is disproportionately up-regulated in.
