exosome-外泌体在肿瘤转移中的作用教学课件

Theroleofexosomesincancermetastasis

外泌体在肿瘤转移中的作用1TheroleofexosomesincExosome1986年,绵羊红细胞上清液中发现了一种有膜结构的小囊泡,命名为外泌体。1996年,B细胞分泌外泌体，可通过MHC激活T细胞；其他APC细胞也可。2013年诺贝尔生物/医学奖2Exosome1986年,绵羊红细胞上清液中发现了一种有膜结Morphology40-100nm圆形双脂质层来源广泛，几乎所有细胞都可分泌分布广泛，血液、尿液、胸水、唾液作用广泛，包含信息丰富3Morphology40-100nm3biogenesis4biogenesis455Exosomalcontent6Exosomalcontent677Biologicalfunctionsandeffectsantigenpresentationimmuneregulationtissuedevelopmentcell-to-cellspreadofinfectiousdiscardsmembraneproteinscancerdevelopment8Biologicalfunctionsandeffec外泌体的临床应用

BiomarkerDrugdelivery9外泌体的临床应用Biomarker9肿瘤转移1.EMT2.Detachment3.circulation4.Adhesion5.Migration6.invasion7.MET8.Engraftment9.Proliferation10.cancerstroma/immunescape10肿瘤转移1.EMT101.exosomesasmediatorsinEMTSnail(zincfingerproteinsSnailandSlug),Zeb(zincfingerandhomeodomainproteinsZeb1and2)andTwist(basichelix-loop-helixproteinsE12,E47,Twist1,Twist2andId)pathways111.exosomesasmediatorsinEMEMTandmigrationTDEsfromtumorcellswhohaveundergoneEMTcaninturnstimulateneighbouringcellstoacquireEMTlikefeatures,creatingasynergisticeffectEBVinfection->nasopharyngealcarcinomacells->exosme->HIF1a->SnailandTwistpathway->moreinvasivephenotypeinrecipientcellsmuscle-invasivebladdercancercells->exosome->urothelialcells->EMTincrease(exosomefromembryonickidneycellsfailed)ExosomalmiR-23a,MiR-191andlet7a12EMTandmigrationTDEsfromtum2.Exosomeand

invasionHighlights•miR-105isuniquelyexpressedandsecretedbymetastaticbreastcancercells•miR-105directlytargetsthetightjunctionproteinZO-1•Cancer-secretedmiR-105destroysendothelialbarriersinthehost•CirculatingmiR-105predictsmetastasisinearly-stagebreastcancerpatients132.Exosomeand

invasionHighlBlockingofastrocyte-exosomesinhibitsbrainmetastasis14Blockingofastrocyte-exosomesExosomesDerivedfromHypoxicOralSquamousCellCarcinomaCellsDelivermiR-21toNormoxicCellstoElicitaPrometastaticPhenotype

15ExosomesDerivedfromHypoxic3.ExosomesandCAFsWefoundthatsomecancer-derivedexosomescouldtriggerelevatedα-smoothmuscleactinexpressionandotherchangesconsistentwiththeprocessoffibroblastdifferentiationintomyofibroblasts.WeshowthatTGF-βisexpressedattheexosomesurfaceinassociationwiththetransmembraneproteoglycanbetaglycan.Althoughexistinginalatentstate,thiscomplexwasfullyfunctionalinelicitingSMAD-dependentsignaling.SolubleTGF-b1aloneisnotabletodrivestromadifferentiationtoacancer-associatedphenotype.Exosome-depletedcancercellsfailtogainastroma-mediatedgrowthadvantageinvivoandtheTGF-b1effectonstromadifferentiationisabrogatedbyblockingofexosomes163.ExosomesandCAFsWefoundtCAFsproduceexosomes,whichstimulatetheWnt-pathwayinbreastcancercellsandenhancetheirmigratorycapabilities17CAFsproduceexosomes,whichs4.EngraftmentofmetastaticcellsCAFsstimulateneoangiogenesisviasecretionofexosomalSDF-1(stromalcellderivedfactor-1)TDEsstimulateendothelialprogenitorcellstoformtube-likestructures.Inductionofneoangiogenesisisnotonlyduetotheexosome’stransportofparacrinesignalingfactorsbutalsoduetodirecttransportofrelevantmRNAtothesurroundingstroma.184.Engraftmentofmetastaticce5.Exosomesinorganotropicmetastaticgrowth1.Apre-metastaticnicheformationisrequiredfortumorcellstoengraftadistantorganOurdatashowthatexosomeproduction,transferandeducationofbonemarrowcellssupportstumorgrowthandmetastasis,hasprognosticvalueandofferspromisefornewtherapeuticdirectionsinthemetastaticprocess.Inaddition,weidentifiedanexosome-specificmelanomasignaturewithprognosticandtherapeuticpotentialcomprisedofTYRP2,VLA-4,HSP70,anHSP90isoformandtheMEToncoprotein.2.195.Exosomesinorganotropicme2020M.Y.Fong,W.Zhou,L.Liu,A.Y.Alontaga,M.Chandra,J.Ashby,etal.,Breast-cancer-secretedmiR-122reprogramsglucosemetabolisminpremetastaticnichetopromotemetastasis,Nat.CellBiol.17(2)(2015)183–194.Hereweshowthatcancercellscansuppressglucoseuptakebynon-tumourcellsinthepremetastaticniche,bysecretingvesiclesthatcarryhighlevelsofthemiR-122microRNA.HighmiR-122levelsinthecirculationhavebeenassociatedwithmetastasisinbreastcancerpatients,andweshowthatcancer-cell-secretedmiR-122facilitatesmetastasisbyincreasingnutrientavailabilityinthepremetastaticniche.Mechanistically,cancer-cell-derivedmiR-122suppressesglucoseuptakebynichecells

in

vitro

and

in

vivobydownregulatingtheglycolyticenzymepyruvatekinase.

In

vivoinhibitionofmiR-122restoresglucoseuptakeindistantorgans,includingbrainandlungs,anddecreasestheincidenceofmetastasis.21M.Y.Fong,W.Zhou,L.Liu,A.22227.Immune-modulatingeffectsofexosomes237.Immune-modulatingeffectsofAcentraleventinvolvesanaberrantexpressionofCOX-2whichinfluencescell-cycleprogressionandcontributetotheacquisitionofacellmigratoryphenotypethroughtheinductionofepithelialmesenchymaltransitiongenesanddown-regulationofE-cadherinexpression.Theidentificationofnovelmoleculardeterminantsinvolvedinthecross-talkbetweenplateletsandcancercellshasledtoidentifynoveltargetsforanti-cancerdrugdevelopment.24Acentraleventinvolvesanab2525TDEsandcellularimmuneresponse1.InaninvitroculturemodelofAML,TDEsdecreasedthecountofCD8+-T-cellsbyactivationofFas/FasL-mediatedapoptosis2.TheyfurtherpromotedCD4+-T-cellproliferationandconversionintoregulatoryT-cellswithincreasedexpressionofIL-10(interleukin-10),TGF-b1,CTLA-4(cytotoxiclymphocyteantigen-4)andGrB(granzymeB).ThesemediatorsreducecytotoxicactivityofNKcells[26TDEsandcellularimmunerespo3.Inaninvitromodelofnasopharyngealcancer,immunosuppressivemiRNAs(hsa-miR-24-3p,hsa-miR-891a,hsa-miR-106a-5p,hsa-miR-20a-5p,andhsa-miR-1908)werefoundinTDEs4.TDEsactivatehumanmyeloidderivedsuppressorcellsthroughHSP72/TLR-2(toll-likereceptor2)viatheSTAT3pathwayandIL-6expression273.InaninvitromodelofnasTDEsandhumoralimmuneresponseneutralizeantitumor-antibodiesInbreastcancercellsitwasdemonstratedthatTDEsexpressHer2andEpCAMantigens,whichbindandneutralizeanti-bodiesinterferingwithADCCoftumorcellsexpressingtheseantigens.Asaconsequence,theseTDEsmayreducethetherapeuticeffectofTrastuzumabAccordingly,B-celllymphomacellssecreteexosomes,whichcarryCD20,neutralizingthetherapeuticeffectofRituximab(anti-CD20-antibody)invivo.28TDEsandhumoralimmuneresponTDEsandcancer-promotingproinflammatoryeffectsTDEscreateaprotumorigenicinflammatoryresponseandsupporttheestablishmentandmaintenanceofapre-metastaticniche.1.TDEsstimulatemacrophagesbyactivatingNF-kB(nuclearfactor-kB).Theseeffectsaremediatedthroughtoll-likereceptors(TLRs)onthesurfaceofmacrophages,whichinteractwithTDEs,especiallyTLR-2.ActivationofmacrophagesbyTLRs(TLR-7and−8)couldbeachievedbybindingofexosomalmiRNAs(mi-R21,mi-R29a2.TDEsofpancreaticductaladenocarcinomacellsinteractwithKupffer-cellsinthelivertopromotepre-metastaticnicheformation29TDEsandcancer-promotingproi总结Exosome在肿瘤转移的各个环节中都有一定作用1.EMT2.Detachment3.circulation4.Adhesion5.Migration6.invasion7.MET8.Engraftment9.Proliferation10.cancerstroma/immunescape30总结Exosome在肿瘤转移的各个环节中都有一定作用30Theroleofexosomesincancermetastasis

外泌体在肿瘤转移中的作用31TheroleofexosomesincExosome1986年,绵羊红细胞上清液中发现了一种有膜结构的小囊泡,命名为外泌体。1996年,B细胞分泌外泌体，可通过MHC激活T细胞；其他APC细胞也可。2013年诺贝尔生物/医学奖32Exosome1986年,绵羊红细胞上清液中发现了一种有膜结Morphology40-100nm圆形双脂质层来源广泛，几乎所有细胞都可分泌分布广泛，血液、尿液、胸水、唾液作用广泛，包含信息丰富33Morphology40-100nm3biogenesis34biogenesis4355Exosomalcontent36Exosomalcontent6377Biologicalfunctionsandeffectsantigenpresentationimmuneregulationtissuedevelopmentcell-to-cellspreadofinfectiousdiscardsmembraneproteinscancerdevelopment38Biologicalfunctionsandeffec外泌体的临床应用

BiomarkerDrugdelivery39外泌体的临床应用Biomarker9肿瘤转移1.EMT2.Detachment3.circulation4.Adhesion5.Migration6.invasion7.MET8.Engraftment9.Proliferation10.cancerstroma/immunescape40肿瘤转移1.EMT101.exosomesasmediatorsinEMTSnail(zincfingerproteinsSnailandSlug),Zeb(zincfingerandhomeodomainproteinsZeb1and2)andTwist(basichelix-loop-helixproteinsE12,E47,Twist1,Twist2andId)pathways411.exosomesasmediatorsinEMEMTandmigrationTDEsfromtumorcellswhohaveundergoneEMTcaninturnstimulateneighbouringcellstoacquireEMTlikefeatures,creatingasynergisticeffectEBVinfection->nasopharyngealcarcinomacells->exosme->HIF1a->SnailandTwistpathway->moreinvasivephenotypeinrecipientcellsmuscle-invasivebladdercancercells->exosome->urothelialcells->EMTincrease(exosomefromembryonickidneycellsfailed)ExosomalmiR-23a,MiR-191andlet7a42EMTandmigrationTDEsfromtum2.Exosomeand

invasionHighlights•miR-105isuniquelyexpressedandsecretedbymetastaticbreastcancercells•miR-105directlytargetsthetightjunctionproteinZO-1•Cancer-secretedmiR-105destroysendothelialbarriersinthehost•CirculatingmiR-105predictsmetastasisinearly-stagebreastcancerpatients432.Exosomeand

invasionHighlBlockingofastrocyte-exosomesinhibitsbrainmetastasis44Blockingofastrocyte-exosomesExosomesDerivedfromHypoxicOralSquamousCellCarcinomaCellsDelivermiR-21toNormoxicCellstoElicitaPrometastaticPhenotype

45ExosomesDerivedfromHypoxic3.ExosomesandCAFsWefoundthatsomecancer-derivedexosomescouldtriggerelevatedα-smoothmuscleactinexpressionandotherchangesconsistentwiththeprocessoffibroblastdifferentiationintomyofibroblasts.WeshowthatTGF-βisexpressedattheexosomesurfaceinassociationwiththetransmembraneproteoglycanbetaglycan.Althoughexistinginalatentstate,thiscomplexwasfullyfunctionalinelicitingSMAD-dependentsignaling.SolubleTGF-b1aloneisnotabletodrivestromadifferentiationtoacancer-associatedphenotype.Exosome-depletedcancercellsfailtogainastroma-mediatedgrowthadvantageinvivoandtheTGF-b1effectonstromadifferentiationisabrogatedbyblockingofexosomes463.ExosomesandCAFsWefoundtCAFsproduceexosomes,whichstimulatetheWnt-pathwayinbreastcancercellsandenhancetheirmigratorycapabilities47CAFsproduceexosomes,whichs4.EngraftmentofmetastaticcellsCAFsstimulateneoangiogenesisviasecretionofexosomalSDF-1(stromalcellderivedfactor-1)TDEsstimulateendothelialprogenitorcellstoformtube-likestructures.Inductionofneoangiogenesisisnotonlyduetotheexosome’stransportofparacrinesignalingfactorsbutalsoduetodirecttransportofrelevantmRNAtothesurroundingstroma.484.Engraftmentofmetastaticce5.Exosomesinorganotropicmetastaticgrowth1.Apre-metastaticnicheformationisrequiredfortumorcellstoengraftadistantorganOurdatashowthatexosomeproduction,transferandeducationofbonemarrowcellssupportstumorgrowthandmetastasis,hasprognosticvalueandofferspromisefornewtherapeuticdirectionsinthemetastaticprocess.Inaddition,weidentifiedanexosome-specificmelanomasignaturewithprognosticandtherapeuticpotentialcomprisedofTYRP2,VLA-4,HSP70,anHSP90isoformandtheMEToncoprotein.2.495.Exosomesinorganotropicme5020M.Y.Fong,W.Zhou,L.Liu,A.Y.Alontaga,M.Chandra,J.Ashby,etal.,Breast-cancer-secretedmiR-122reprogramsglucosemetabolisminpremetastaticnichetopromotemetastasis,Nat.CellBiol.17(2)(2015)183–194.Hereweshowthatcancercellscansuppressglucoseuptakebynon-tumourcellsinthepremetastaticniche,bysecretingvesiclesthatcarryhighlevelsofthemiR-122microRNA.HighmiR-122levelsinthecirculationhavebeenassociatedwithmetastasisinbreastcancerpatients,andweshowthatcancer-cell-secretedmiR-122facilitatesmetastasisbyincreasingnutrientavailabilityinthepremetastaticniche.Mechanistically,cancer-cell-derivedmiR-122suppressesglucoseuptakebynichecells

in

vitro

and

in

vivobydownregulatingtheglycolyticenzymepyruvatekinase.

In

vivoinhibitionofmiR-122restoresglucoseuptakeindistantorgans,includingbrainandlungs,anddecreasestheincidenceofmetastasis.51M.Y.Fong,W.Zhou,L.Liu,A.52227.Immune-modulatingeffectsofexosomes537.Immune-modulatingeffectsofAcentraleventinvolvesanaberrantexpressionofCOX-2whichinfluencescell-cycleprogressionandcontributetotheacquisitionofacellmigratoryphenotypethroughtheinductionofepithelialmesenchymaltransitiongenesanddown-regulationofE-cadherinexpression.Theidentificationofnovelmoleculardeterminantsinvolvedinthecross-talkbetweenplateletsandcancercellshasledtoidentifynoveltargetsforanti-cancerdrugdevelopment.54Acentraleventinvolvesanab5525TDEsandcellularimmuneresponse1.InaninvitroculturemodelofAML,TDEsdecreasedthecountofCD8+-T-cellsbyactivationofFas/FasL-mediatedapoptosis2.TheyfurtherpromotedCD4+-T-cellproliferationandconversionintoregulatoryT-cellswithincreasedexpressionofIL-10(interleukin-10),TGF-b1,CTLA-4(cytotoxiclymphocyteantigen-4)andGrB(granzymeB).ThesemediatorsreducecytotoxicactivityofNKcells[56TDEsandcellularimmunerespo3.Inaninvitromodelofnasopharyngealcancer,immunosuppressivemiRNAs(hsa-miR-24-3p,hsa-miR-891a,hsa-miR-106a-5p,hsa-miR-20a-5p,andhsa-miR-1908)werefoundin