肿瘤分子生物学9. 肿瘤微的环境张志刚文档资料.ppt

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1、1889 种子与土壤假说1909 免疫监视1910 病毒与肿瘤1915 激素与癌症1937 肿瘤干细胞1939 血管再生术1950 吸烟与肿瘤1953 二次突变假说1960 染色体易位1971 肿瘤抑制基因 细胞凋亡与肿瘤1975 肿瘤微环境1976克隆演变&多步骤肿瘤发生,1976病毒癌基因的细胞同系物 1978 癌基因编码 蛋白调控细胞生长 1979 第一个人类癌基因 1983 癌基因互作、肿瘤表观遗传学 1989 细胞周期和DNA损伤检查1990 肿瘤易感性的遗传基础 肿瘤遗传不稳定性的机制 1999 肿瘤谱 2001 肿瘤靶向治疗,肿瘤研究百年大事记,（Nature Milestone

2、s in Cancer,milestones of timelines),1889 种子与土壤假说(Seed and Soil Hypothesis),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

3、to have 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.,Molecu

4、lar mechanism(until today),?,1975 肿瘤微环境(Tumor Microenvironment),Beatricemintz and Karl Illmensee,They 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-der

5、ived cells were present in large numbers and contributed to several unrelated tissues,the most notable being functional spermatozoa.They concluded tumor cells were developmental totipotent and could revert to normal behavior in the appropriate environment.,（Nature,milestones of timelines),1984 肿瘤微环境

6、(Tumor Microenvironment),Mina Bissell,Context is everything,People and idea,JCB,2009,The 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 th

7、at if RSV was injected into 4-day-old embryo,no tumor were produced,despite the spread of RSV infection and active v-src expression.,Other contributions:,The wounding is important for tumorigenesis,Blocking integrin function is sufficient to revert the maglignant phenotype of human breast cancer.,Ge

8、neral concept of tumor microenvironment,Tumor microenvironment refers to the 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

9、 variable,with differences seen between patients and often in different areas 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:,主要特征:,Tumor microenvironment,Tumor cell,Tumor-associated str

10、omal cells（fibroblast，endothelial cell,Immune cell）,Extracellular Matrix Proteins（structural and non-structural),Acidic,Cytokines,Neurotransmitters,Hypoxia,Growth factors,MMPs,ADAMs,other secreted proteins and their receptors,Hormone,Chemokines,Tumor-associated stromal cells,Activated fibroblast(hep

11、atic stellate cell),Immune cell,Endothelial cell,Pericyte,Adipocyte,Glial cell,Tumor associated fibroblast,Characteristics of normal fibroblast,Activated fibroblast in the tumor microenvironment,Origin of activated fibroblasts in the tumor stroma,Interaction between tumor cells and activated fibrobl

12、asts,The role of activated fibroblast in cancer development,Cross-talk between activated fibroblasts and other tumor-associated host cell,Characteristics of normal fibroblast,Basic component of connective tissue and contribute to structural integrity,Main resource of ECM and regulation of ECM turnov

13、er,Low proliferate index and minimum metabolic capacity,Main resource of growth factors and cytokines,Activated fibroblast in the tumor microenvironment,Similar phenotype as fibroblasts in inflammation and wound healing,Indented nuclei,stress fibers and well-developed cell-matrix interaction,Markers

14、:Fibroblast activation protein(FAP),a-smooth muscle actin(aSMA),r-smooth muscle actin(gSMA),Prolyl-4-hydroxylase(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：,如何鉴定：,Origin of

15、activated fibroblasts in the tumor stroma,Surrounding resting fibroblasts after their activation 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 biology,Interaction betwee

16、n tumor cells and activated fibroblasts,Tumor,Activated fibroblasts,TGF-b,PDGF,LL-37,LPA,All the stages of tumor development,The role of activated fibroblast in cancer development,Initiation of tumor,Kuperwasser,et al.,2004,PNAS,Bhnowmick,et al.,2004,Science,Progression of tumor,Bhnowmick,et al.,200

17、4,Science,Metastasis of tumor,Grugan,et al.,2010,PNAS,Duda,et al.,2010,PNAS,TGF-beta signaling in fibroblasts modulates 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.,Con

18、ditional knock-out TGF-beta type II receptor,Fibroblasts,Abundance of stromal cells,Intraepithelial neoplasia in prostate（5-7 weeks）,Invasive squamous cell carcinoma of the forestomach（7-week-old）,Paracrine HGF signaling,（FSP1 promotor）,3 weeks,Initiation of tumor,Fibroblast-secreted hepatocyte grow

19、th factor plays a functional role in esophageal squamous cell carcinoma invasion.Grugan KD,Miller CG,Yao Y,Michaylira CZ,Ohashi 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 signaling,Invasion of tumor,Ma

20、lignant cells facilitate lung metastasis by bringing their own soilDuda DG,Duyverman AM,Kohno M,Snuderl M,Steller EJ,Fukumura D,Jain RK Proc Natl Acad Sci U S A.2010 Dec 14;107(50):21677-82,Metastasis of tumor,Cross-talk between myofibroblasts and other tumor-associated host cell,Wever et al.2008,In

21、t.J.Cancer,Suppression of antitumor immunity by stromal cells expressing fibroblast activation protein-alpha.Kraman M,Bambrough PJ,Arnold JN,Roberts EW,Magiera L,Jones JO,Gopinathan A,Tuveson DA,Fearon DT.Science.2010 Nov 5;330(6005):827-30.,Transgenic mouse,FAP(fibroblast activation protein-a)promo

22、tor,Expressing Primate diphtheria toxin receptor,Endothelial cells and tumor,Tumor angiogenesis:Therapeutic implication.N.Engl.J.Med.,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 s

23、ecrete a soluble factor that stimulates the proliferation of endothelial cells,that these in turn control tumour expansion and,in the 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 canc

24、er progression.Franses JW,Baker AB,Chitalia VC,Edelman ER.Sci Transl Med.2011 Jan 19;3(66):66ra5,Secretions from quiescent endothelial cell,(EC-conditioned medium),Breast and lung carcinoma proliferation and invasiveness,Knockdown of perlecan,Eliminates EC ability to suppress cancer invasiveness,Inh

25、ibit,Tumor microenvironment,Tumor cell,Tumor-associated stromal cells（fibroblast，endothelial cell,Immune cell）,Extracellular Matrix Proteins（structural and non-structural),PH,Cytokines,Neurotransmitters？,Hypoxia,Growth factors,MMPs,ADAMs,other secreted proteins and their receptors,Extracelluar Matri

26、x,The extracellular matrix(ECM)the extracellular part of animal tissue that usually provides structural support to the animal cells in addition to performing various other important functions.The extracellular matrix is the defining feature of connective tissue in animals.,Extracellular matrix inclu

27、des the interstitial matrix and the basement membrane.,Interstitial matrix:present between various animal cells,Basement membrane:sheet-like depositions of ECM on which various epithelial cells rest,Composition of ECM,Proteins,Collagens,Laminins,-G-X-Y-,a,b,g subunit,Fibronectin,Elastins,Proteoglyca

28、ns(蛋白聚糖）,Syndecan(硫酸乙酰肝素蛋白聚糖),1.Classical extracellular matrix,2.Matricellular proteins,Osteopontin(OPN),Osteonectin(SPARC),thrombospondins,tenascin,CCNs,Biology of Collagens,Structure(Triple helix),Basic Composition,-Glycine-Proline-Hydroxyproline(hydroxylysine)-,Types of Collagen,29 types of colla

29、gens have been found,Collagens is the most abundant protein in mammals,making up about 25%to 35%of the whole-body protein content,Usage,Cosmetic,Medical,Laminins:ubiquitous proteins of basement membranes,Epithelia,Vasculature,Laminins:a family of proteins,Specific spatio-temporal expression 111embry

30、onic 211,221mainly muscles411,421mainly endothelium332mainly epithelium511,512more ubiquitous,Aumailley et al.,A simplified laminin nomenclatureMatrix Biol.,2005,24,326-332,5 a,3 b,3 g subunits,16 laminins,Subunits of laminins,LAMA1:3075 amino acids,337084 Da;17 EGF-like domain,5 LamG domain;1 LamNT

31、 domain,2 LamB domain.,a subunit,LAMB1:1786 amino acids,198066 Da;13 EGF-like domain,1 LamNT domain,1 PDB domain(Protein Data Bank,chemotaxis).,b subunit,LAMC1:1609 amino acids,177603 Da;10 EGF-like domain,1 LamNT domain,1 LamB domain,1 Scop domain(Structural Classification of Proteins,Apolipophorin

32、-III).,g subunit,Roles of ECM in Biology,Providing support and anchorage for cells,Segregating tissues from one another,Regulating intercellular communication,Reservoir of growth factors,ECM and ECM fragments can function as growth factor,ECM in pathological processes,Wound healing,Fibrosis,Cancer B

33、iology,Invasion,Metastasis,Tumor Progression,Inflammation,Extracelluar 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 cancer,Metastatic tumor cells,Escape

34、 from anoikis and invade other organs,Cancer stem cell,Extracelluar Matrix,A Matter of Life and Death,Pro SURVIVAL ECM,Fibronectin prosurival,2386 amino acids;262625 Da,FNI domain,FNII domain,FNIII domain,15 FNIII domain,RGD motif for integrins and other protein interaction,Confer resistance to apop

35、tosis;activates ERK1/2,ROCK and PI3K/AKT pathway,Pro SURVIVAL ECM,Fibronectin prosurival,Fibronectin,a5b1 integrin,FAK,ERK,RhoA,Cell survival,Pro SURVIVAL ECM,Rescues cells from apoptosis by protein kinase B/AKT activation,Laminin 10/11,a5b1g1,a5b2g1,Laminin 10/11,a3b1 integrin,p130cas,PI3K/AKT,Rac1

36、,Cell survival,Extracelluar Matrix,A Matter of Life and Death,ECM stiffness(Physical properties),Leads to integrin clusterization and triggers mitogenic signals,Physico-mechanical aspects of extracellular matrix influences on tumorigenic behaviors.Cukierman E,Bassi DE.Semin Cancer Biol.2010 Jun;20(3

37、):139-45.,ECM stiffness influence tumor initiation,progression and metastasis,From fibrosis to cancer,Roles of ECM in cancer,Matrix crosslinking forces tumor progression by enhancing integrin signaling,Levental et al.,2009,Cell,This article offer a new paradigm for understanding why tumor incidence

38、so dramatically with aging.,ECM stiffness on celluar behaviors,Extracelluar Matrix,A Matter of Life and Death,Pro APOPTOTIC ECM,CCN1,Induces apoptosis through a6b1 integrin and syndecan-4 binding,CCN family(6 members):cysteine-rich protein 61(Cyr61 or CCN1)connective tissue growth factor(CTGF or CCN

39、2)nephroblastoma(肾母细胞瘤)overexpressed protein(Nov or CCN3)Wnt-inducible secreted protein-1(WISP-1 or CCN4)WISP-2(CCN5)WISP-3(CCN6),SPARC,Induces apoptosis through a yet unknown mechanism,Decorin,Induces apoptosis through activation of caspase-3 by an unknown mechanism,EMILIN2,Induces apoptosis throug

40、h death receptors DR4 and DR5 direct binding,Pro APOPTOTIC ECM,Endostatin,Induces apoptosis through downregulation of apoptosis inhibitors,EMILIN2(Elastin Microfibril Interface Located Protein 2),Matricellular Proteins,At the crossroad of infalmmation and cancer,Matricellular proteins are a class of

41、 non-structural ECM protein exerting regulative functions likely through a direct binding to specific surface receptors,to growth factors and cytokines,to other matrix proteins.,Matricellular Proteins,At the crossroad of infalmmation and cancer,OPN(Osteopontin),Wound healing,Affected phases during h

42、ealing,Healing rate,Matrix deposition and contraction,Accelerated,Inflammation,Role in inflammation,Leukocyte function,Pro-inflammatory Th1 committing,Pro-survival of lymphocytes,monocytes and neutrophils,Tumorigenesis,Role in tumor-host interaction,Effect on tumor growth,Pro-angiogenic,Involved in

43、tumor progression and angiogenesis.Level of expression correlated with stages of disease(breast and prostate),Matricellular Proteins,At the crossroad of infalmmation and cancer,TSP-1,Wound healing,Affected phases during healing,Healing rate,TGFb activationinflammation,In transgenic mice overexpressi

44、on TSP-1,Inflammation,Role in inflammation,Leukocyte function,Resolving inflammation,Recruits monocyte and neutrophilsNegative regulator of DC and Th1 effectors activation neutrophils chearance,Tumorigenesis,Role in tumor-host interaction,Effect on tumor growth,Stromal TSP-1 is anti-angiogenic,Tumor

45、 growth,Angiogenesis and activates TGFb,Healing in also in TSP-1 KO mice,Matricellular Proteins,At the crossroad of infalmmation and cancer,TSP-2,Wound healing,Affected phases during healing,Healing rate,Collagen deposition,Accelerated,Inflammation,Role in inflammation,Leukocyte function,Inflammatio

46、n and angiogenesis in TSP-2 KO mice,Tumorigenesis,Role in tumor-host interaction,Effect on tumor growth,Stromal TSP-2 is anti-angiogenic,TSP-2 deficiency susceptibility to skin carcinogenesis andtumor formation,Roles of ECM in cancer,Desmoplasia,Appears at relatively early-stage lesions and disappea

47、rs in highly invasive,advanced tumor,It could be a host defense reaction designed to confine the developing tumor by tumor associated with stromal cells,Fragments of ECM in Angiogenesis,Roles of ECM in Angiogenesis,Collagen XVIII,inhibits angiogenesis,C-terminal fragment,Endostatin,Trimeric NC1 doma

48、in,promotes angiogenesis,1 NC domain,1 FRI(Frizzled)domain,1 LamG,6 Collagen doamains,1 Endostatin,Roles of Endostatin in Cancer,Perlecan,Promote tumor growth angiogenesis,endorepellin,C-terminal fragment from domain V of Perlecan,inhibits angiogenesis,Roles of ECM in Angiogenesis,Roles of ECM in ca

49、ncer invasion and metastasis,Roles of ECM in cancer invasion and metastasis,Laminin 332,Laminin 332 g2 chain has been found at leading edge of invading carcinomas,Bladder,Breast,Cervical,Colorectal,Gastric,Head and neck,Liver,Lung,Pancreas,prostate and many others,Osteopontin,Versican,Prognostic fac

50、tor,proliferation,adhesion,motility,metastasis,angiogenesis,Promotion of metastasis,inhibition of apoptosis,neoangiogenesis,Carcinoma-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