肿瘤分子生物学9. 肿瘤微的环境张志刚文档资料

《肿瘤分子生物学9. 肿瘤微的环境张志刚文档资料》由会员分享，可在线阅读，更多相关《肿瘤分子生物学9. 肿瘤微的环境张志刚文档资料（83页珍藏版）》请在装配图网上搜索。

1、肿瘤研究百年大事记肿瘤研究百年大事记（Nature Milestones in Cancer,milestones of timelines)Stephen Paget(assistant surgeon to the West London hospital)By analyzing 735 case histories of fatal breast cancer,he found that metastasis formed in the liver far moreeven those such as the spleen that could be considered to hav

2、e the same exposure to the cancer cells because of similar blood flows.（Nature,milestones of timelines)Question:“what is it that decides what organ shall suffer a case of disseminated cancer”Conclusion:Cancer cells require some“nourishment nourishment”from the environment to develop.Molecular mechan

3、ism(until today)?Beatricemintz and Karl IllmenseeThey took the tetracarcinoma cells from embryoid bodies in vivo,and injected them into developing mouse blastocysts.Surprisingly,normal mice were born with no evidence of tumors.They found that tumor-derived cells were present in large numbers and con

4、tributed to several unrelated tissues,the most notable being functional spermatozoa.They c o n c l u d e d t u m o r c e l l s w e r e developmental totipotent and could revert to normal behavior in the appropriate environment.（Nature,milestones of timelines)Mina BissellContext is everythingPeople a

5、nd idea,JCB,2009The tumor-inducing behavior of Rous sarcoma virus(RSV)when injected into the wings of newly hatched chicks was already known,and the viral gene v-src had been identified as the sole culprit.What Bissell found was that if RSV was injected into 4-day-old embryo,no tumor were produced,d

6、espite the spread of RSV infection and active v-src expression.Other contributions:The wounding is important for tumorigenesisBlocking integrin function is sufficient to revert the maglignant phenotype of human breast cancer.General concept of tumor microenvironmentTumor microenvironment refers to t

7、he unique properties of tissue microenvironment conferred by abnormal interactions between tumor and host cells.Tumor microenvironment is often characterized by hypoxia,nutrient deprivation,acidosis,and aberrant stroma.Highly variable,with differences seen between patients and often in different are

8、as of the same tumor.The tumor microenvironment is often altered as the diseases progresses;even the percentage of a tumor made up of cancer cells may change.基本定义基本定义:主要特征主要特征:Tumor microenvironmentTumor cellTumor-associated stromal cells（fibroblast，endothelial cell,Immune cell）Extracellular Matrix

9、Proteins（structural and non-structural)AcidicCytokinesNeurotransmitters HypoxiaGrowth factorsMMPs,ADAMs,other secreted proteins and their receptors HormoneChemokinesTumor-associated stromal cellsActivated fibroblast(hepatic stellate cell)Immune cellEndothelial cellPericyteAdipocyteGlial cellTumor as

10、sociated fibroblastCharacteristics of normal fibroblastActivated fibroblast in the tumor microenvironmentOrigin of activated fibroblasts in the tumor stromaInteraction between tumor cells and activated fibroblastsThe role of activated fibroblast in cancer developmentCross-talk between activated fibr

11、oblasts and other tumor-associated host cellCharacteristics of normal fibroblastBasic component of connective tissue and contribute to structural integrityMain resource of ECM and regulation of ECM turnoverLow proliferate index and minimum metabolic capacityMain resource of growth factors and cytoki

12、nesActivated fibroblast in the tumor microenvironmentSimilar phenotype as fibroblasts in inflammation and wound healingIndented nuclei,stress fibers and well-developed cell-matrix interactionMarkers:Fibroblast activation protein(FAP),a-smooth muscle actin(aSMA),r-smooth muscle actin(gSMA),Prolyl-4-h

13、ydroxylase(P4H,for collagen synthesis)Identification:a-SMA in combination with at least 3 other markers(e.g.,positive for a-SMA,vimentin and P4H and negative for cytokeratin)基本特征：基本特征：如何鉴定：如何鉴定：Origin of activated fibroblasts in the tumor stromaSurrounding resting fibroblasts after their activation

14、and recruitment to the tumor tissue,Bone marrow stem cell,pericytes,via epithelial or endothelial to mesenchymal transition.Xouri,et.al.2010,seminars in cell and developmental biologyInteraction between tumor cells and activated fibroblastsTumorActivated fibroblastsTGF-b,PDGF,LL-37,LPAAll the stages

15、 of tumor developmentThe role of activated fibroblast in cancer developmentInitiation of tumorKuperwasser,et al.,2004,PNASBhnowmick,et al.,2004,ScienceProgression of tumorBhnowmick,et al.,2004,ScienceMetastasis of tumorGrugan,et al.,2010,PNASDuda,et al.,2010,PNASTGF-beta signaling in fibroblasts mod

16、ulates the oncogenic potential of adjacent epithelia.Bhowmick NA,Chytil A,Plieth D,Gorska AE,Dumont N,Shappell S,Washington MK,Neilson EG,Moses HL.Science.2004 Feb 6;303(5659):848-51.Conditional knock-out TGF-beta type II receptorFibroblastsAbundance of stromal cellsIntraepithelial neoplasia in pros

17、tate（5-7 weeks）Invasive squamous cell carcinoma of the forestomach（7-week-old）Paracrine HGF signaling（FSP1 promotor）3 weeksInitiation of tumorFibroblast-secreted hepatocyte growth factor plays a functional role in esophageal squamous cell carcinoma invasion.Grugan KD,Miller CG,Yao Y,Michaylira CZ,Oh

18、ashi S,Klein-Szanto AJ,Diehl JA,Herlyn M,Han M,Nakagawa H,Rustgi AK.Proc Natl Acad Sci U S A.2010 Jun 15;107(24):11026-31.Epub 2010 Jun 1.HGF/MET signalingInvasion of tumorMalignant cells facilitate lung metastasis by bringing their own soilDuda DG,Duyverman AM,Kohno M,Snuderl M,Steller EJ,Fukumura

19、D,Jain RK Proc Natl Acad Sci U S A.2010 Dec 14;107(50):21677-82 Metastasis of tumorCross-talk between myofibroblasts and other tumor-associated host cellWever et al.2008,Int.J.CancerSuppression of antitumor immunity by stromal cells expressing fibroblast activation protein-alpha.Kraman M,Bambrough P

20、J,Arnold JN,Roberts EW,Magiera L,Jones JO,Gopinathan A,Tuveson DA,Fearon DT.Science.2010 Nov 5;330(6005):827-30.Transgenic mouseFAP(fibroblast activation protein-a)a)promotorExpressing Primate diphtheria toxin receptorEndothelial cells and tumorTumor angiogenesis:Therapeutic implication.N.Engl.J.Med

21、.1971 Judah Folkman In 1939,Gordon Ide and colleagues In 1945,Glenn Algire and colleagues In 1968,Melvin Greenblatt and Philippe Shubik He proposed that tumour cells secrete a soluble factor that stimulates the proliferation of endothelial cells,that these in turn control tumour expansion and,in the

22、 absence of new vessel growth,that tumours do not increase beyond 23 mm in size,entering instead a state of dormancy.Stromal endothelial cells directly influence cancer progression.Franses JW,Baker AB,Chitalia VC,Edelman ER.Sci Transl Med.2011 Jan 19;3(66):66ra5Secretions from quiescent endothelial

23、cell(EC-conditioned medium)Breast and lung carcinoma proliferation and invasivenessKnockdown of perlecanEliminates EC ability to suppress cancer invasivenessInhibitTumor microenvironmentTumor cellTumor-associated stromal cells（fibroblast，endothelial cell,Immune cell）Extracellular Matrix Proteins（str

24、uctural and non-structural)PHCytokinesNeurotransmitters？HypoxiaGrowth factorsMMPs,ADAMs,other secreted proteins and their receptors Extracelluar Matrix The extracellular matrix(ECM)the extracellular part of animal tissue that usually provides structural support to the animal cells in addition to per

25、forming various other important functions.The extracellular matrix is the defining feature of connective tissue in animals.Extracellular matrix includes the interstitial matrix and the basement membrane.Interstitial matrix:present between various animal cellsBasement membrane:sheet-like depositions

26、of ECM on which various epithelial cells restComposition of ECMProteinsCollagensLaminins-G-X-Y-a,b,g subunitFibronectinElastinsProteoglycans(蛋白聚糖）蛋白聚糖）Syndecan(硫酸乙酰肝素蛋白聚糖蛋白聚糖)1.Classical extracellular matrix 2.Matricellular proteins Osteopontin(OPN),Osteonectin(SPARC),thrombospondins,tenascin,CCNsBi

27、ology of CollagensStructure(Triple helix)Basic Composition-Glycine-Proline-Hydroxyproline(hydroxylysine)-Types of Collagen29 types of collagens have been foundCollagens is the most abundant protein in mammals,making up about 25%to 35%of the whole-body protein content UsageCosmetic,MedicalLaminins:ub

28、iquitous proteins of basement membranesEpitheliaVasculatureMuscleGlandNerveLaminins:a family of proteins111,121211,221,213511,521,523a1,a2,a5 g g1,g g3b b1,b b2a3A a4 g g1,g g3b b1,b b2311,321411,421,423a3Ag g2b b3332Aa3Bg g2b b3332BSpecific spatio-temporal expression 111embryonic 211,221mainly musc

29、les411,421mainly endothelium332mainly epithelium511,512more ubiquitousAumailley et al.,A simplified laminin nomenclatureMatrix Biol.,2005,24,326-3325 a a,3 b b,3 g g subunits,16 lamininsSubunits of lamininsLAMA1:3075 amino acids,337084 Da;17 EGF-like domain,5 LamG domain;1 LamNT domain,2 LamB domain

30、.a a subunitLAMB1:1786 amino acids,198066 Da;13 EGF-like domain,1 LamNT domain,1 PDB domain(Protein Data Bank,chemotaxis).b b subunitLAMC1:1609 amino acids,177603 Da;10 EGF-like domain,1 LamNT domain,1 LamB domain,1 Scop domain(Structural Classification of Proteins,Apolipophorin-III).g g subunitRole

31、s of ECM in Biology Providing support and anchorage for cells Segregating tissues from one another Regulating intercellular communication Reservoir of growth factorsECM and ECM fragments can function as growth factorECM in pathological processesWound healingFibrosisCancer BiologyInvasionMetastasisTu

32、mor ProgressionInflammationExtracelluar Matrix A Matter of Life and Death Anoikis is a form of programmed cell death which is induced by anchorage-dependent cells detaching from the surrounding extracellular matrix(ECM).Anoikis and cancerMetastatic tumor cellsEscape from anoikis and invade other org

33、ans Cancer stem cellExtracelluar Matrix A Matter of Life and Death Pro SURVIVAL ECMFibronectin prosurival2386 amino acids;262625 Da FNI domain,FNII domain,FNIII domain15 FNIII domain,RGD motif for integrins and other protein interactionConfer resistance to apoptosis;activates ERK1/2,ROCK and PI3K/AK

34、T pathwayPro SURVIVAL ECMFibronectin prosurivalFibronectina a5b b1 integrinFAKERKRhoACell survivalPro SURVIVAL ECMRescues cells from apoptosis by protein kinase B/AKT activationLaminin 10/11a a5b b1g g1,a a5b b2g g1Laminin 10/11a a3b b1 integrinp130casPI3K/AKTRac1Cell survivalExtracelluar Matrix A M

35、atter of Life and Death ECM stiffness(Physical properties)Leads to integrin clusterization and triggers mitogenic signalsPhysico-mechanical aspects of extracellular matrix influences on tumorigenic behaviors.Cukierman E,Bassi DE.Semin Cancer Biol.2010 Jun;20(3):139-45.ECM stiffness influence tumor i

36、nitiation,progression and metastasisFrom fibrosis to cancerFriends or foeRoles of ECM in cancerFriends or foeMatrix crosslinking forces tumor progression by enhancing integrin signalingLevental et al.,2009,Cell This article offer a new paradigm for understanding why tumor incidence so dramatically w

37、ith aging.ECM stiffness on celluar behaviorsExtracelluar Matrix A Matter of Life and Death Pro APOPTOTIC ECMCCN1Induces apoptosis through a6b1 integrin and syndecan-4 bindingCCN family(6 members):cysteine-rich protein 61(Cyr61 or CCN1)connective tissue growth factor(CTGF or CCN2)nephroblastoma(肾母细胞瘤

38、)overexpressed protein(Nov or CCN3)Wnt-inducible secreted protein-1(WISP-1 or CCN4)WISP-2(CCN5)1.WISP-3(CCN6)SPARCInduces apoptosis through a yet unknown mechanismDecorinInduces apoptosis through activation of caspase-3 by an unknown mechanismEMILIN2Induces apoptosis through death receptors DR4 and

39、DR5 direct bindingPro APOPTOTIC ECMEndostatinInduces apoptosis through downregulation of apoptosis inhibitorsEMILIN2(Elastin Microfibril Interface Located Protein 2)Matricellular ProteinsAt the crossroad of infalmmation and cancerMatricellular proteins are a class of non-structural ECM protein exert

40、ing regulative functions likely through a direct binding to specific surface receptors,to growth factors and cytokines,to other matrix proteins.Matricellular ProteinsAt the crossroad of infalmmation and cancerOPN(Osteopontin)Wound healingAffected phases during healingHealing rateMatrix deposition an

41、d contractionAcceleratedInflammationRole in inflammationLeukocyte functionPro-inflammatory Th1 committingPro-survival of lymphocytes,monocytes and neutrophilsTumorigenesisRole in tumor-host interactionEffect on tumor growthPro-angiogenicInvolved in tumor progression and angiogenesis.Level of express

42、ion correlated with stages of disease(breast and prostate)Matricellular ProteinsAt the crossroad of infalmmation and cancerTSP-1Wound healingAffected phases during healingHealing rateTGFb activationinflammationIn transgenic mice overexpression TSP-1InflammationRole in inflammationLeukocyte functionR

43、esolving inflammationRecruits monocyte and neutrophilsNegative regulator of DC and Th1 effectors activation neutrophils chearanceTumorigenesisRole in tumor-host interactionEffect on tumor growthStromal TSP-1 is anti-angiogenicTumor growth,Angiogenesis and activates TGFb Healing in also in TSP-1 KO m

44、iceMatricellular ProteinsAt the crossroad of infalmmation and cancerTSP-2Wound healingAffected phases during healingHealing rateCollagen depositionAcceleratedInflammationRole in inflammationLeukocyte functionInflammation and angiogenesis in TSP-2 KO miceTumorigenesisRole in tumor-host interactionEff

45、ect on tumor growthStromal TSP-2 is anti-angiogenicTSP-2 deficiency susceptibility to skin carcinogenesis andtumor formationRoles of ECM in cancerFriends or foeDesmoplasia Appears at relatively early-stage lesions and disappears in highly invasive,advanced tumor It could be a host defense reaction d

46、esigned to confine the developing tumor by tumor associated with stromal cells Fragments of ECM in AngiogenesisRoles of ECM in AngiogenesisCollagen XVIIIinhibits angiogenesisC-terminal fragment,EndostatinTrimeric NC1 domainpromotes angiogenesis1 NC domain,1 FRI(Frizzled)domain,1 LamG,6 Collagen doam

47、ains,1 EndostatinRoles of Endostatin in CancerPerlecanPromote tumor growth angiogenesisendorepellinC-terminal fragment from domain V of Perlecaninhibits angiogenesisRoles of ECM in AngiogenesisRoles of ECM in cancer invasion and metastasisRoles of ECM in cancer invasion and metastasisLaminin 332 Lam

48、inin 332 g g2 chain has been found at leading edge of invading carcinomasBladder,Breast,Cervical,Colorectal,Gastric,Head and neck,Liver,Lung,Pancreas,prostate and many othersOsteopontinVersicanPrognostic factor,proliferation,adhesion,motility,metastasis,angiogenesisPromotion of metastasis,inhibition

49、 of apoptosis,neoangiogenesisCarcinoma-produced factors activate myeloid cells through TLR2 to stimulate metastasis.Kim S,Takahashi H,Lin WW,Descargues P,Grivennikov S,Kim Y,Luo JL,Karin M.Nature.2009 Jan 1;457(7225):102-6.VersicanTN-a，IL-6MetastasisECM as a circulating marker for cancer diagnosisTu

50、mor MarkerMain Application Field1.Mucins,proteoglycans,glycoproteinsEpisialin and related(MUC-1 family,CA15-3,CA 549,CAM29,CAM26,CASA,BCM,KL-6,MCA,TAG72)Sialyl Lewis and related(CA 19-9,GICA,CA 195)Cancer antigen 125 and related related(CA125,CA602,OVX)TRA-1-60Soluble glypican-3Soluble mesothelinCar

51、cino-embryonic antigen and related(CEA,NCA)2.Host-derived mediatorsOsteoprotegerin,bone alkaline phosphatase3.Soluble ECM breakdwon productsLaminin fragmentsCollagen type IVType I Cllagen Carboxyterminal Telopeptide(ICTP)Circulating ECM complexes(DR-70)Breast carcinomaPancreatic,bile duct and colore

52、ctal carcinomaOvarian carcinomaTesticular germ cell tumor(no-seminous)MelanomaMesotheliomaColorectal carcinomaSkeletal metastasis(prostatic cancer)Colorectal carcinomaBreast carcinomaSkeletal metastasis(prostatic cancer)Lung carcinomasheet-like depositions of ECM on which various epithelial cells re

53、stBasement membraneBMBasement membrane structureUltrastructureKeratinocyteBasementmembraneKeratinsBP230PlectinCD151CollagenXVIICollagenIVInterstitialcollagensCollagenVIILM332a a6b b4CellmembraneDermis3070 nanometers in thicknessBMMMPsBMBMInvasion through the basement membraneCellproliferation+increa

54、singintra-tumoralpressure=disruptionof BMTumor cellextendsfilopodiaandmigratetowardsdermalsignalDiffusiblemediatorDermalalterationThe receptor of ECMIntegrin familyNon-integrin family18 a a subunit,8 b b subunit,more than 20 integrinsSyndecan,a a-dystroglycan,67 kD laminin receptor(Laminin receptors

55、)DDRs,GPVI,LAIR-1,uPARAP/Endo180(collagen receptors)Integrin activation4 subfamily integrins47E5683b10125891473611Collagen-binding integrinLaminin-bindingintegrinsRGD-bindingintegrinsL2MXDLeukocyteintegrinsECM receptors-integrin familyRoles of Integrins in cancerRegulation of cell survival and apopt

56、osisRoles of Integrins in cancerRegulation of angiogenesisRoles of Integrins in cancerRegulation of migration and invasionRoles of Integrins in cancerRegulation of cancer stem cellIntegrins interaction with receptors of growth factorVEGF Receptor (Ruoslahti 2002)TGFB Receptor (Scaffidi et al.2004)c-

57、MET(HGF receptor)(Sridhar&Miranti 2006)PDGF Receptor(Sridhar&Miranti 2006)EGF Receptor(Bill et al.2006)Type-1 insulin-like growth factor receptor(Bill et al.2006)Insulin Receptor(Schneller et al.1997)Integringrowth factor and integrincytokine receptor crosstalkIntegrins in the host defense to cancer

58、Integrins as targets for cancer therapyRoles of MMP in cancerMatrix metalloproteinases(MMPs)are zinc-dependent endopeptidasesCapable of degrading all kinds of extracellular matrix proteins,but also can process a number of bioactive moleculesInvolved in the cleavage of cell surface receptors,the rele

59、ase of apoptotic ligands(such as the FAS ligand),and chemokine/cytokine in/activationFunctionsProliferationMigrationDifferentiationAngiogenesisApoptosisHost defenseRoles of MMP in cancer(A family of zinc-dependent endopeptidases)MMPs have been implicated in Cancer for more than 40 yearsMMP-mediated

60、ECM degradation leads to cancer cell invasion and metastasis has been a guiding pricipleNew insights into the Roles of MMPs in CancerMMPs affect growth signalsMMPs regulate apoptosisRegulation of tumor vasculatureInitiation of Neoplastic progressionTissue invasion and metastasisMetastatic niche form

61、ationOrchestrate inflammation in cancerNew insights into the Roles of MMPs in CancerMatrix metalloproteinases:regulators of the tumor microenvironment.Kessenbrock K,Plaks V,Werb Z.Cell.2010 Apr 2;141(1):52-67.ReviewMMPs and tumor microenvironmentOther proteinases in ECM turnoverADAMs(A Disintegrin A

62、nd Metalloproteases)inflammation,cancer,Alzheimers particularly in tumor-stroma interactionTumor microenvironmentTumor cellTumor-associated stromal cells（fibroblast，endothelial cell,Immune cell）Extracellular Matrix Proteins（structural and non-structural)AcidicCytokinesNeurotransmitters？HypoxiaGrowth

63、 factorsMMPs,ADAMs,other secreted proteins and their receptors Metastasis and angiogenesis induced by hypoxiaThe process of metastasis often occurs in regions of hypoxia,in which HIF can induce expression of pro-invasive and pro-angiogenic factors,such as LOX,CTGF,VEGF,Ang-2 and decreases in E-cadhe

64、rin and thrombospondin.Tumor acidic microenvironment OxygenNutrientDeprivationAbnormal metabolism(Warburg effect)GlycolysisLactic acidacidic microenvironmentChemoresistanceNormal cell deathECM degradation and cancer metastasis德国生物化学家,曾获1931年诺贝尔生理学-医学奖 Neurotransmitters and cancerSystem regulation of cancer microenvironmentBMECMIntegrinsCytokinesActivated fibroblastImmune cellChemokinesNeurotransmitterEpithelial cellCarcinoma Growth factor神经系统神经系统免疫系统免疫系统内分泌系统内分泌系统Together!Together!We can beat cancer!