1、分子细胞生物学进展分子细胞生物学进展Metabolic Strategies in Cancer癌症的代谢策略癌症的代谢策略Lecture Objectivesn To provide a brief summary of the history of research in cancer metabolismn To summarize the metabolic strategies in cancer cellsn To reveal genetic mechanisms behind the metabolic phenotypes in tumorsn To give an exam
2、ple of transcriptional regulation in cancer metabolism(our work on ChREBP)TermsGlycolysis 糖酵解糖酵解TCA cycle/Oxidative phosphorylation 三羧酸循环三羧酸循环/氧化磷酸化氧化磷酸化Macromolecules（生物）大分子（生物）大分子Anabolism 合成代谢合成代谢Catabolism 分解代谢分解代谢Isoenzyme 同工酶同工酶Warburg effect/aerobic glycolysis 有氧糖酵解有氧糖酵解FDG-PET 18F脱氧葡萄糖（脱氧葡萄糖
3、（FDG）-正电子发射体层显像（正电子发射体层显像（PET）Hallmarks of cancer 癌症的特征癌症的特征IDH(isocitrate dehydrogenase)异柠檬酸脱氢酶异柠檬酸脱氢酶Oncometabolite 致癌代谢物致癌代谢物Metabolic syndrome 代谢综合症代谢综合症Truncated TCA cycle 截短的截短的/不完整的不完整的三羧酸循环三羧酸循环ReferencesThe Molecular Basis of Cancer(Third Edition)Copyright 2008 Elsevier Inc.All rights reser
4、ved Edited by:John Mendelsohn,MD,Peter M.Howley,MD,Mark A.Israel,MD,Joe W.Gray,PhD,and Craig B.Thompson,MD ISBN:978-1-4160-3703-3Understanding the Warburg effect:the metabolic requirements of cell proliferation.Vander Heiden MG,Cantley LC,Thompson CB.Science.2009 May 22;324(5930):1029-33.Hallmarks o
5、f cancer:the next generation.Hanahan D,Weinberg RA.Cell.2011 Mar 4;144(5):646-74.Metabolic reprogramming:a cancer hallmark even warburg did not anticipate.Ward PS,Thompson CB.Cancer Cell.2012 Mar 20;21(3):297-308.Part In To provide a brief summary of the history of research in cancer metabolismn To
6、summarize the metabolic strategies in cancer cellsn To reveal genetic mechanisms behind the metabolic phenotypes in tumorsn To give an example of transcriptional regulation in cancer metabolism(our work on ChREBP)Otto Warburg and Warburg EffectOtto Heinrich Warburg(1883-1970)Nobel Prize in Physiolog
7、y or Medicine(1931)discovery of the nature and mode of action of the respiratory enzyme.Vander Heiden et al,Science 2009葡萄糖葡萄糖丙酮酸丙酮酸乳酸乳酸氧化氧化磷酸化磷酸化无氧无氧糖酵解糖酵解有氧有氧糖酵解糖酵解FDG-PET(fluorodeoxyglucose-positron emission tomography)Vander Heiden et al,Science 2009静脉注射18F-FDG后，18F-FDG 随血液循环到达肿瘤所在，由于肿瘤细胞表面存在高表达
8、的葡萄糖转运蛋白（Glut1和Glut3）,使18F-FDG大量进入肿瘤细胞内，并在细胞内的已糖激酶（大多数恶性肿瘤内已糖激酶含量明显高于良性病变和正常组织）作用下，转变为6-磷酸-18F-FDG,由于FDG与正常葡萄糖存在分子结构的差异，它不参与葡萄糖的进一步代谢而滞留在细胞。通过PET动、静态扫描，能准确地测量肿瘤的葡萄糖代谢异常程度并对其进行显像。在某些肿瘤，如高分化原发性肝癌，细胞内存在较高水平的葡萄糖-6-磷酸酶，能将存在于细胞内的6-磷酸-18F-FDG水解成18F-FDG，后者通过自由扩散游离出细胞，18F-FDG便不在肿瘤细胞内聚集而出现假阴性。FDG-PET肿瘤显像的原理肿瘤
9、显像的原理Sidney Weinhouse(1909-2001)isozyme in hepatomas Peter PedersenJohns Hopkins Univ.School of MedicineChi DangUniv.of PennsylvaniaSchool of MedicineGregg SemenzaJohns Hopkins Univ.School of MedicineScientists in the Field of Cancer MetabolismLewis CantleyHarvard Medical SchoolTak MakCanadaUniv.of
10、Toronto管坤良管坤良复旦大学复旦大学UCSD熊跃熊跃复旦大学复旦大学Univ.of North CarolinaAnd many moreCraig ThompsonMemorial Sloan-Kettering Cancer Center 赵世民赵世民复旦大学复旦大学雷群英雷群英复旦大学医学复旦大学医学院院SU2C Dream Team ScientistsTargeting PI3K in Womens CancersLewis C.Cantley,PhD,Charles Sawyers,MD,and Gordon B.Mills,MD,PhDCutting Off the Fue
11、l Supply:A New Approach to the Treatment of Pancreatic CancerCraig Thompson,MD,and Daniel D.von Hoff,MD,FACP Bioengineering and Clinical Applications of Circulating Tumor Cell ChipDaniel Haber,MD,PhD,and Mehmet Toner,PhDBringing Epigenetic Therapy to the Forefront of Cancer ManagementStephen Baylin,
12、MD,and Peter A.Jones,PhDAn Integrated Approach to Targeting Breast Cancer Molecular Subtypes and Their Resistance Phenotypes Joe Gray,PhD,and Dennis Slamon,MD,PhDThe Original Six Hallmarks of CancerHanahan D and Weinberg RA,Cell.2000生长信号的生长信号的自给自足性自给自足性对于抑制生长信号对于抑制生长信号的不敏感性的不敏感性逃避细胞凋亡逃避细胞凋亡无限的复制潜力无限
13、的复制潜力持续的血管生成持续的血管生成组织浸润和转移组织浸润和转移基因组不稳定基因组不稳定和突变和突变细胞能量异常细胞能量异常促进肿瘤的炎症促进肿瘤的炎症避免免疫摧毁避免免疫摧毁The New Ten Hallmarks of CancerHanahan D and Weinberg RA,Cell.2011n 在分子水平上抑制代谢中的关键酶而降低糖酵解和其它合成代谢通路活性可以减缓肿瘤细胞生长。n 对癌症病人全基因组测序找到了某些高频率突变的代谢基因，这些基因的突变导致代谢中间物失调从而对于细胞表观遗传性状和信号转导通路产生重要影响。n 大规模流行病学研究表明肥胖病，糖尿病等代谢性综合症患者
14、的各类癌症发病率明显上升。Why is Cancer Metabolism Important?n 在分子水平上抑制代谢中的关键酶而降低糖酵解和其它合成代谢通路活性可以减缓肿瘤细胞生长。n 对癌症病人全基因组测序找到了某些高频率突变的代谢基因，这些基因的突变导致代谢中间物失调从而对于细胞表观遗传性状和信号转导通路产生重要影响。n 大规模流行病学研究表明肥胖病，糖尿病等代谢综合征患者的各类癌症发病率明显上升。Why is Cancer Metabolism Important?GARBER K,Science 2006HK=Hexokinase,己糖激酶PDK=pyruvate dehydrog
15、enase kinase,丙酮酸脱氢酶激酶PDH=pyruvate dehydrogenase,丙酮酸脱氢酶ACL=ATP citrate lyase,ATP-柠檬酸裂解酶Strategies for Targeting Tumor Energy Pathways(I)Strategies for Targeting Tumor Energy Pathways(II)n Evidence for an alternative glycolytic pathway in rapidly proliferating cells.Vander Heiden MG et al,Science.2010
16、 Sep 17;329(5998):1492-9.n The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth.Christofk HR and Vander Heiden MG et al,Nature.2008 Mar 13;452(7184):230-3.n Pyruvate kinase M2 is a phosphotyrosine-binding protein.Christofk HR andVander Heiden MG et al,Nature.
17、2008 Mar 13;452(7184):181-6.PKM2,pyruvate kinase M2,丙酮酸激酶-M2PGAM,phosphoglyceromutase,磷酸甘油酸变位酶Cancer cell proliferation can be inhibited by decreasing glycolysis and anabolic metabolismGlucoseGlucoseLactateFatty acidAmino acidNucleotideTCAcycleCancer cell proliferationGlucoseGlucoseLactateFatty acid
18、Amino acidNucleotideTCAcycleCancer cell proliferationn 在分子水平上抑制代谢中的关键酶而降低糖酵解和其它合成代谢通路活性可以减缓肿瘤细胞生长。n 对癌症病人全基因组测序找到了某些高频率突变的代谢基因，这些基因的突变导致代谢中间物失调从而对于细胞表观遗传性状和信号转导通路产生重要影响。n 大规模流行病学研究表明肥胖病，糖尿病等代谢综合征患者的各类癌症发病率明显上升。Why is Cancer Metabolism Important?Roles of IDH1,IDH2,SDH,and Fumarase in Cellular Metabol
19、ismIDH,异柠檬酸脱氢酶异柠檬酸脱氢酶SDH,琥珀酸脱氢酶琥珀酸脱氢酶Fumarase,富马酸酶富马酸酶Thompson CB,N Engl J Med.2009 An integrated genomic analysis of human glioblastoma multiforme.神经胶质瘤Parsons DW et al,Science.2008 Sep 26;321(5897):1807-12.IDH1 and IDH2 mutations in gliomas.Yan H et al,N Engl J Med.2009 Feb 19;360(8):765-73.Glioma
20、-derived mutations in IDH1 dominantly inhibit IDH1 catalytic activity and induce HIF-1alpha.Zhao S et al,Science.2009 Apr 10;324(5924):261-5.Cancer-associated IDH1 mutations produce 2-hydroxyglutarate.2-羟基戊二酸Dang L et al,Nature.2009 Dec 10;462(7274):739-44.Leukemic IDH1 and IDH2 mutations result in
21、a hypermethylation phenotype,disrupt TET2 function,and impair hematopoietic differentiation.Figueroa ME et al,Cancer Cell.2010 Dec 14;18(6):553-67.Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of-ketoglutarate-dependent dioxygenases.Xu W et al,Cancer Cell.2011 Jan 18;19(1):17-30.IDH m
22、utation impairs histone demethylation and results in a block to cell differentiation.Lu C et al,Nature.2012 Feb 15;483(7390):474-8.IDH,异柠檬酸脱氢酶异柠檬酸脱氢酶Metabolic pathways provide substrates for post-translational modifications(PTMs)that influence cell signalingMetallo CM et al,Genes Dev.2010n 在分子水平上抑制代
23、谢中的关键酶而降低糖酵解和其它合成代谢通路活性可以减缓肿瘤细胞生长。n 对癌症病人全基因组测序找到了某些高频率突变的代谢基因，这些基因的突变导致代谢中间物失调从而对于细胞表观遗传性状和信号转导通路产生重要影响。n 大规模流行病学研究表明肥胖病，糖尿病等代谢综合征患者的各类癌症发病率明显上升。Why is Cancer Metabolism Important?中华医学会糖尿病学分会（CDS）建议代谢综合征的诊断标准具备以下4项组成成分中的3项或全部者：n 超重和（或）肥胖n 高血糖n 高血压n 血脂紊乱Metabolic SyndromeOverweight,obesity and cance
24、r:epidemiological evidence and proposed mechanisms.Calle EE,Kaaks R.Nat Rev Cancer.2004 Aug;4(8):579-91.Part IIn To provide a brief summary of the history of research in cancer metabolismn To summarize the metabolic strategies in cancer cellsn To reveal genetic mechanisms behind the metabolic phenot
25、ypes in tumorsn To give an example of transcriptional regulation in cancer metabolism(our work on ChREBP)What do Cells Need to Grow and Proliferate?Substrates for Growth(such as Glucose and Glutamine)ATP and NADH and NADPH Appropriate Regulation of Metabolic Pathways to Maximize BiosynthesisMetaboli
26、c Strategies in Cancer CellsCancer cells High nutrient uptakeHigh glycolytic rateIncreased anabolism Low activity of TCA cycleor“Truncated”TCA Cycle.Normal cells Basal nutrient uptakeBasal glycolytic rateBasal anabolism High activity of TCA cycleOtto Warburg and Warburg EffectOtto Heinrich Warburg(1
27、883-1970)Nobel Prize in Physiology or Medicine(1931)discovery of the nature and mode of action of the respiratory enzyme.Vander Heiden et al,Science 2009葡萄糖葡萄糖丙酮酸丙酮酸乳酸乳酸氧化氧化磷酸化磷酸化无氧无氧糖酵解糖酵解有氧有氧糖酵解糖酵解Why is the Rate of Glycolysis So High in Tumor Cells?Reason 1:To Produce Sufficient Energy and Metabo
28、lic Intermediates for BiosynthesisGlycolysis Produces Energy and Metabolic Intermediates for Tumor Cell ProliferationMacromolecules:ProteinLipidNucleic acidProteinLipidNucleic acidWhy is the Rate of Glycolysis So High in Tumor Cells?Reason 2:To Survive Periods of Hypoxia 缺氧缺氧Reason 3:To Secret Lacta
29、te and Create a Microenvironment Favorable for Tumor GrowthTumors Use a“Truncated”Form of the TCA CycleGlutamine Oxidation Makes the Truncated TCA Cycle Possible.Part IIIn To provide a brief summary of the history of research in cancer metabolismn To summarize the metabolic strategies in cancer cell
30、sn To reveal genetic mechanisms behind the metabolic phenotypes in tumorsn To give an example of transcriptional regulation in cancer metabolism(our work on ChREBP)TermsPI3K=Phosphatidylinositol 3-kinases Akt/PKB,a serine/threonine protein kinase P53,a transcription factorAMPK=AMP-activated protein
31、kinase HIF=Hypoxia-inducible factor,a transcription factorMyc,a transcription factorChREBP=carbohydrate responsive element-binding protein,a transcription factor ROS=Reactive oxygen species 活性氧活性氧The phosphatidylinositol 3-kinases(PI3Ks)are members of a conserved family of intracellular lipid kinase
32、s.The PI3K PathwayEngelman JA et al,Nat Rev Genet.2006Mutations in the PI3K Pathway in CancersEngelman JA et al,Nat Rev Genet.2006 The PI3K/Akt Pathway Drives Growth MetabolismP53,a transcription factor.Nutrient Deprivation Signals to p53Vousden KH and Ryan KM.Nat Rev Cancer.2009 Regulation of Energ
33、y Production by p53Vousden KH and Ryan KM.Nat Rev Cancer.2009 Kinase LKB1 and AMPK are activated by the high AMP/ATP ratio that occurs during energy limitation.The LKB1/AMPK Pathway Allows Cells to Respond to Metabolic Stress by Engaging Catabolic MetabolismAMPK=AMP-activated protein kinase The tran
34、scription factor HIF-1 contributes to the high rate of glycolysis in tumors.Semenza GL.Curr Opin Genet Dev.2010HIF-1:Upstream and Downstream of Cancer MetabolismThe Myc proteins are a family of transcription factors that regulate cell cycle entry and metabolism.c-Myc and HIF Regulate Carbon Metaboli
35、smGordan JD et al,Cancer Cell.2007.Myc Regulates both Glucose and Glutamine Metabolic PathwaysDang CV.Cancer Res.2010.GLS=glutaminase,谷氨酰胺酶ASCT2=谷氨酰胺载体Part IVn To provide a brief summary of the history of research in cancer metabolismn To summarize the metabolic strategies in cancer cellsn To reveal
36、 genetic mechanisms behind the metabolic phenotypes in tumorsn To give an example of transcriptional regulation in cancer metabolism(our work on ChREBP)Cancer cell proliferation can be inhibited by decreasing glycolysis and anabolic metabolismGlucoseGlucoseLactateFatty acidAmino acidNucleotideTCAcyc
37、leCancer cell proliferationGlucoseGlucoseLactateFatty acidAmino acidNucleotideTCAcycleCancer cell proliferationMouse ChREBP Protein1864S196S568T666PKAAMPKPKAGlucose sensing module ChREBP(carbohydrate responsive element binding protein)binds the carbohydrate response element(ChoRE)which consists of t
38、wo E-box like consensus sequences separated by five base pairs.In adults,expression of ChREBP is detectable in most tissues but is at its highest levels in liver and adipose tissue.ChREBP null mice show decreased glycolysis and lipogenesis as well as intolerance to dietary carbohydrate.Loss of ChREB
39、P in leptin null ob/ob mice results in alleviation of obesity.NES NLSPro-richbHLH/ZIPZIP likeRoles of LXR,ChREBP,and SREBP-1c in the Transcriptional Control of Triglyceride Synthesis in LiverDenechaud PD et al,JCI 2008Hormone/nutrient-mediated Regulation of ChREBP Activity in HepatocytesUyeda K et a
40、l,Cell metabolism 2006The Role of ChREBP in Glycolysis and Lipogenesis in HepatocytesUyeda K et al,Cell metabolism 2006The question:What is the Role of ChREBP in Cancer Cells?Proliferative Cells Depend on ChREBP to Maintain Proliferative Expansion(HCT116 Colorectal Cancer Cells)Tong et al,Proc Natl
41、Acad Sci U S A.2009 Proliferative Cells Depend on ChREBP to Maintain Proliferative Expansion(HepG2 Cells)Suppression of ChREBP Leads to a Metabolic Switch from Aerobic Glycolysis to Mitochondrial Respiration(HCT116 Colorectal Cancer Cells)Warburg EffectVander Heiden et al,Science 2009Suppression of
42、ChREBP Leads to Decreased Lipid and Nucleotide Biosynthesis(HCT116 Cells)DeBerardinis RJ et al,Proc Natl Acad Sci U S A.2007Effect of ChREBP Knockdown on Labeling in Acid Soluble Metabolites from 1,6-13C2Glucose(HCT116 Cells)ControlChREBP knockdownpro-4lac-3ala-3glut-4glth-4glut-3glut-2Glycolytic In
43、termediatesasp-3HHHHHHgn-2gn-4gn-3pro-4lac-3glut-4glut-3glut-2Glycolytic Intermediatesasp-3HHHHHHglut-2/glut-4=0.230.04glut-2/glut-4=0.370.07Suppression of ChREBP Induces p53 Activation(HCT116 Cells)Suppression of ChREBP Induces Cell Cycle Arrest(HCT116 Cells)Nutrient Deprivation Signals to p53Vousd
44、en KH and Ryan KM.Nat Rev Cancer.2009 Suppression of ChREBP Does not Activate AMPKMurphy,Biochemical Journal 2009Overview of Mitochondrial ROS ProductionIncreased ROS Level and Decreased Cellular Glutathione Level in ChREBP-deficient Cells Compared with Control Cells(HCT116 Cells)Antioxidant N-acety
45、lcysteine(NAC)Reduces ChREBP Knockdown-induced p53 Activation(HCT116 Cells)Suppression of ChREBP Leads to a p53-dependent Reduction in Tumor Growth ChREBP is highly expressed in liver and colorectal cancer cells and critical for cancer cell proliferationGlucoseGlucoseLactateFatty acidAmino acidNucle
46、otideTCAcycleHigh ChREBP expressioncancer cell proliferationGlucoseGlucoseLactateFatty acidAmino acidNucleotideTCAcycleLow ChREBP expressioncancer cell proliferationNutrients(glucose,amino acid,fatty acid,lactate,etc.)ChREBPMondoAMondo familyGlycolysis,anabolic metabolism and proliferation in colore
47、ctal and liver cancer cellsOur group studies transcriptional regulation in cancer cell metabolismCancer cell linesMouse modelsClinical samplesLecture Objectivesn To provide a brief summary of the history of research in cancer metabolismn To summarize the metabolic strategies in cancer cellsn To reveal genetic mechanisms behind the metabolic phenotypes in tumorsn To give an example of transcriptional regulation in cancer metabolism(our work on ChREBP)
