Targeting Glucose Metabolism in Cancer

靶向癌症中的葡萄糖代谢

• 批准号: 8462570
• 负责人: Sucheta Telang
• 金额: $ 21.42万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8462570
• 关键词: 6-Phosphofructo-2-kinase; 6-Phosphofructokinase; A549; Actins; Adenocarcinoma Cell; Age-Months; Anchorage-Independent Growth; Apoptosis; Attenuated; Basement membrane; Binding; Brain; Breast; Carbon; Cell Fractionation; Cells; Chickens; Citric Acid Cycle; Colon; Cytoplasm; Data; Development; Enzymes; Epithelial Cells; Exons; Family; Freezing; Fructose; Gel; Genes; Genomics; Glucose; Glycolysis; Glycolysis Inhibition; Growth; Heart; Human; Hypoxia; Immunohistochemistry; In Situ; In Situ Nick-End Labeling; In Vitro; Investigation; Isoenzymes; Knockout Mice; Label; Lactic acid; Lamins; Large T Antigen; Liver; Lung; Lung Adenocarcinoma; Lung Neoplasms; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Measures; Messenger RNA; Metabolic; Metabolism; Mitochondria; Modeling; Mouse Mammary Tumor Virus; Mus; Mutation; NADH; Normal tissue morphology; Nude Mice; Oncogenic; Open Reading Frames; Organ; Ovary; Oxygen; Oxygen Consumption; Pathway interactions; Phenotype; Phosphoric Monoester Hydrolases; Phosphotransferases; Placenta; Positron-Emission Tomography; Production; Protein p53; Proteins; Regulation; Relative (related person); Reporting; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Site; Skeletal Muscle; Small Interfering RNA; Source; Specimen; Subfamily lentivirinae; TOCSY; Tamoxifen; Telomerase; Testing; Testis; Tissues; Transgenes; Tumor Tissue; Vascularization; Western Blotting; base; cancer cell; cancer type; cell transformation; cellular transduction; design; glucose metabolism; glucose uptake; in vivo; inhibitor/antagonist; mRNA Expression; mortality; neoplastic; neoplastic cell; neurotensin mimic 1; novel; overexpression; promoter; protein expression; public health relevance; recombinase; screening; shunt pathway; small hairpin RNA; small molecule; tumor; tumor growth; vector

DESCRIPTION (provided by applicant): A high rate of glycolytic flux, even in the presence of oxygen, is a central metabolic hallmark of neoplastic tumors. Cancer cells preferentially utilize glycolysis in order to satisfy their increased energetic and biosynthetic requirements. This metabolic phenotype is confirmed by positron emission tomography (PET) with 2-[18F]-fluoro-2-deoxy-glucose which demonstrates that tumors take up 10-fold more glucose than adjacent normal tissues in vivo. Over-expression of HIF-1a and myc, ras activation and loss of p53 function stimulate glycolysis by activating a family of four bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases (PFKFB) which synthesize fructose-2,6-bisphosphate (F2,6BP), an allosteric activator of 6-phosphofructo-1- kinase (PFK-1) which is an essential control point in the glycolytic pathway. The PFKFB enzymes are encoded by four genes (PFKFB1-4) reportedly expressed in distinct tissues: liver/skeletal muscle (PFKFB1), heart (PFKFB2), placenta (PFKFB3) and testis (PFKFB4). Previous studies have focused on the PFKFB3 isozyme as the dominant source of F2,6BP in neoplastic cells due to its high kinase:phosphatase ratio (~740:1) and overexpression in multiple tumors. The identification of the specific PFKFB isozyme responsible for the high rate of glycolysis in cancer cells may allow for the development of novel agents that suppress tumor growth through inhibition of glycolysis. In preliminary studies, we measured mRNA expression of the four PFKFB isozymes in 20 tumor tissues by multiplex RT-PCR and found that PFKFB4 mRNA (and not PFKFB3) was markedly increased in 17/20 tumors relative to adjacent, normal tissues. We speculated that the high PFKFB4 expression may be caused by disparate oncogenic pathways converging to enhance glycolytic flux. We found that hypoxic exposure of K-rasG12S+ A549 lung adenocarcinoma cells and introduction of H-rasG12V into immortalized human bronchial epithelial cells increased PFKFB4 mRNA and protein expression. Further, cell fractionation revealed that the PFKFB4 protein, and not PFKFB3, localized to the cytoplasm, the cellular compartment of both PFK-1 and glycolysis. Last, transient siRNA silencing of PFKFB4 mRNA expression in K- rasG12S+ A549 cells decreased steady-state concentration of F2,6BP, glycolytic flux and anchorage- independent tumor growth in mice. Taken together, these preliminary studies provide rationale for further investigation of the PFKFB4 isozyme as a possible novel target for the development of anti-neoplastic agents. We hypothesize that the PFKFB4 isozyme is required for the high glycolytic flux, survival, growth and spread of transformed cells. We plan to test this hypothesis by pursuing the following specific aims: 1. To examine the requirement for F2,6BP synthesized by PFKFB4 for glycolytic and mitochondrial metabolism. 2. To determine the requirement for F2,6BP synthesized by PFKFB4 for growth, invasiveness and survival of normal, immortalized and ras-transformed epithelial cells. 3. To examine the effect of PFKFB4 genomic deletion on growth and metabolism of ras-dependent tumors in vivo using a Cre-lox inducible mouse knockout of PFKFB4.

描述（由申请人提供）：即使在存在氧气的情况下，糖酵解通量的高速率也是肿瘤性肿瘤的中心代谢标志。癌细胞优先利用糖酵解以满足其增加的能量和生物合成需求。这种代谢表型通过2-[18 F]-氟-2-脱氧葡萄糖的正电子发射断层扫描（PET）证实，这表明肿瘤在体内摄取的葡萄糖比邻近正常组织多10倍。HIF-1a和myc的过表达、ras的激活和p53功能的丧失通过激活一个由四种双功能6-磷酸果糖-2-激酶/果糖-2，6-二磷酸酶（PFKFB）组成的家族来刺激糖酵解，所述PFKFB合成果糖-2，6-二磷酸（F2，6 BP），所述F2，6 BP是糖酵解途径中的重要控制点6-磷酸果糖-1-激酶（PFK-1）的变构激活剂。PFKFB酶由四种基因（PFKFB 1 -4）编码，据报道在不同的组织中表达：肝脏/骨骼肌（PFKFB 1）、心脏（PFKFB 2）、胎盘（PFKFB 3）和睾丸（PFKFB 4）。以前的研究集中在PFKFB 3同工酶作为肿瘤细胞中F2，6 BP的主要来源，因为其高激酶：磷酸酶比（~740：1）和在多种肿瘤中的过表达。鉴定负责癌细胞中糖酵解的高速率的特异性PFKFB同工酶可以允许开发通过抑制糖酵解来抑制肿瘤生长的新型药剂。在初步研究中，我们测量了四个PFKFB同工酶的mRNA表达在20个肿瘤组织的多重RT-PCR，发现PFKFB 4的mRNA（而不是PFKFB 3）显着增加，在17/20肿瘤相对于相邻的正常组织。我们推测PFKFB 4的高表达可能是由于不同的致癌途径会聚在一起以增强糖酵解通量。我们发现低氧暴露K-rasG 12 S + A549肺腺癌细胞和将H-rasG 12 V引入永生化的人支气管上皮细胞增加PFKFB 4 mRNA和蛋白表达。此外，细胞分级显示PFKFB 4蛋白而不是PFKFB 3定位于细胞质，即PFK-1和糖酵解的细胞区室。最后，K-rasG 12 S + A549细胞中PFKFB 4 mRNA表达的瞬时siRNA沉默降低了小鼠中F2、6 BP的稳态浓度、糖酵解通量和锚定非依赖性肿瘤生长。综上所述，这些初步研究提供了进一步调查的PFKFB 4同工酶作为一个可能的新的目标，用于开发抗肿瘤药物的基本原理。我们推测PFKFB 4同工酶是转化细胞的高糖解通量、存活、生长和扩散所必需的。我们计划通过追求以下具体目标来测试这一假设：1。研究PFKFB 4合成的F2，6 BP在糖酵解和线粒体代谢中的需要。2.确定正常、永生化和ras转化上皮细胞生长、侵袭和存活对PFKFB 4合成的F2，6 BP的需求。3.使用Cre-lox诱导的PFKFB 4敲除小鼠，检查PFKFB 4基因组缺失对ras依赖性肿瘤体内生长和代谢的影响。

项目成果

6-Phosphofructo-2-kinase (PFKFB3) promotes cell cycle progression and suppresses apoptosis via Cdk1-mediated phosphorylation of p27.

• DOI: 10.1038/cddis.2014.292
• 发表时间: 2014-07-17
• 期刊: Cell death & disease
• 影响因子: 9

Fructose-2,6-bisphosphate synthesis by 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4 (PFKFB4) is required for the glycolytic response to hypoxia and tumor growth.

6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶 4 (PFKFB4) 合成果糖-2,6-二磷酸是对缺氧和肿瘤生长的糖酵解反应所必需的。

• DOI: 10.18632/oncotarget.2213
• 发表时间: 2014-08-30
• 期刊: Oncotarget
• 作者: Chesney J;Clark J;Klarer AC;Imbert-Fernandez Y;Lane AN;Telang S
• 通讯作者: Telang S

Targeting the sugar metabolism of tumors with a first-in-class 6-phosphofructo-2-kinase (PFKFB4) inhibitor.

• DOI: 10.18632/oncotarget.4534
• 发表时间: 2015-07-20
• 期刊: Oncotarget
• 作者: Chesney J;Clark J;Lanceta L;Trent JO;Telang S
• 通讯作者: Telang S

Inhibition of 6-phosphofructo-2-kinase (PFKFB3) induces autophagy as a survival mechanism.

• DOI: 10.1186/2049-3002-2-2
• 发表时间: 2014-01-23
• 期刊: Cancer & metabolism
• 影响因子: 5.9
• 作者: Klarer AC;O'Neal J;Imbert-Fernandez Y;Clem A;Ellis SR;Clark J;Clem B;Chesney J;Telang S
• 通讯作者: Telang S