靶向Jagged1-依赖性肿瘤-基质相互作用在乳腺癌症骨转移中的作用

TargetingJagged1-DependentTumor-StromalInteractionsinBreastCancerBoneMetastasisDepartmentofMolecularBiology,PrincetonUniversityYibinKangBonemetastasisOsteolyticBoneMetastasisofBreastCancerNormalhttp://surgical-.uk

OsteoclastsBreastCancerCellsBoneOsteoblast/OsteocytesTheViciousCycleinBoneMetastasisEll&Kang,Cell,2012OsteoclastResorptionFormationBoneRANKLOPGOsteoblastTumorderivedfactorsTGF

IGFCa2+Korpaletal,NatureMedicine,2009Bone-deriveTGFFeedsBacktoTumorCellsP-Smad2stainingKorpaletal,NatureMedicine,2009SCP3SCP4SCP6SCP2SCP25SCP46IL11CTGFSKILSGKJUNBANGPTL4SPHK1JAG1CDKN1APLAURankinordereddatasetofbonemetastasisEnrichmentscore(ES)Coregeneset0.100.20.30.4Jagged1isOneoftheTGF-ResponsiveGenesintheBoneMetastasisGeneSetTGFβ(h)Jagged1β-actin061224Jagged1Delta-like1Delta-like4Delta-like1Jagged1Jagged267NR4T0766c144T1MetastasisabilityJagged1isAssociatedwithIncreasedRiskofBoneMetastasisJagged1ProbesSCP4SCP6SCP39ATCCSCP25SCP46SCP20SCP2WeakStrongBoneMetastaticActivity%BoneMet-freesurvivalYears05100255075100Logrankp=0.0327Jagged1Low(n=41)Jagged1High(n=41)Minnetal.dataset,82patientsinMSKCCBA-+-+ControlJagged1-KDTGFbJagged1

-actinADaysafterTumorInjectionBoneMetastasisBLI11010210310471421283542SCP2-ControlSCP2-Jagged1KD****Jagged1KnockdownInhibitsOsteolyticBoneMetastasisBControlJagged1KD05101520P=0.022ControlKDOsteolyticarea(mm2)C#TRAP+osteoclasts/mmboneinterfaceBBTTVectorJagged1KDTRAP01020304050P=0.007ControlKDSethi,etal,CancerCell,2011Jagged1-OverexpressionStimulatesTumorGrowthinCo-CulturewithOsteoblastsN1ICDHes1,Hey1….CSLNucleusSignal-sendingCellNotch1Jagged1

-Secretase

Complex

GSISignal-receivingCellN1ICD050100150200250300350Colonydiameter(μM)

DMSOGSIControlJagged1******ControlJagged1ControlJagged1DMSOGSI20x5xGFP/Luc+TumorCell+/-Jagged1MC3T3OsteoblastNotch1+Jagged1-OverexpressingTumorCellsStimulatesOsteoclastogenesisGFP/Luc+TumorCell+/-Jagged1RAW264.7Pre-osteoclastNotch1+TRAP+Osteoclasts05101520****DMSOGSIControlJagged1ControlJagged1DMSOGSIControlJagged1ControlJagged1TRAPStainingOsteoclastprogenitorBoneTGF

RI/IIOsteoblastTumorCellTumor-StromalCrosstalkinBoneMetastasisbyMediatedbyTGF

andNotchPathwaysOsteoclastJagged1/Notch1T

RinhibitorsGSI,Jagged1AbTGF

IL-6,CTGFNFATc171421283542BoneMetastasisBurden1101021030.1104Control+VehicleJagged1+Vehicle*DaysafterTumorInjectionNotchSignalingInhibitorGSIInhibitsJagged1-MediatedBoneMetastasisVector+GSI**Jagged1+GSISCP28ControlJagged1ControlJagged1VehicleGSIBBBBH&ETRAPBLIX-rayTTTTH&EAlcianblueControlJag1Ab(15D11)GSI(MRK-003)NoSignificantGITrackToxicityofJag1AbgobletcellmetaplasiaDay1:TumorXenograftDay0:15D1110mg/kg15D11or3mg/kgOPG,twiceweeklyfor5weeksCTIgGJag1AbOPG-FcSCP28-Jagged1OsteoclastsX-rayHealthyBoneSCP28-Jag1+OPG*SCP28-Jag1+Jag1Ab**SCP28-Jag1+IgGBoneMetastasisBurdenWeeksafterTumorInjection110102103104234561Jagged1AntibodyInhibitsJagged1-MediatedBoneMetastasisinPreventiveTreatmentProtocol10mg/kgJag1Ab,twiceweeklyfor4weeksDay1:TumorxenograftNotreatmentfor2weeksNoSignificantBenefitinLateTreatmentProtocol0501001502002345NormalizedBLISignalIgGJag1AbOPGN.S.SCP28-Jag1Weeks10mg/kgJag1Ab,twiceweeklyfor3weeksDay1:TumorxenograftNotreatmentfor1weeksCombinedJag1Ab/OPGTreatmentInhibitstheGrowthofEstablishedBoneMetastasis01002003004005006001234IgGJag1AbOPGJag1Ab+OPGP=0.02NormalizedBLIsignalIgGJag1AbOPGJag1Ab+OPGWeeksDay7Day280123451234567WeeksNormalizedBLI(log)IgGPaclitaxolJag1AbPaclitaxol+Jag1Ab6JaggedAbSensitizesBoneMetastasistoChemotherapyDay1:ICinjectionOneweeklater:AborchemotreatmentTwiceaweekWeek7:TerminalanalysisIgGPaclitaxolJag1AbPaclitaxol

+Jag1AbSCP28-Jag1TheStromalRoleofJagged1?Jagged1expressionissignificantlyelevatedinbonestromalcellsduringfactureandrepair,particularlyinMSCsandosteoblasts.Jagged1ishighlyexpressedinbonestromalcellsinmetastaticbonelesions,andisfurtherinducedinresponsetosystemictherapy.Notchsignalingisimportantforbonedevelopment.SkeletalDevelopmentDefectsofCola1-Jagged1TransgenicMiceCol1a1-Jag1WTCol1a1-Jag1WT***WTCol1a-Jag102004006008008008509009501000BD(mgHA/ccm)***50100150WTCol1a-Jag10TRAP+cells200WTCol1a-Jag1012345#Ob/BSWTCol1a-Jag101234#Oc/BS**IncreasedOsteoclastActivityinCola1-Jagged1TransgenicMiceWT

Col1a1-Jag1CtskInvivoInvitroIgGIgGJag1AbJag1AbWTCol1a1-Jag1Col1a1-Jag1Jag1AbCol11-Jag1

IgG

AbJag1AbTreatmentReversesBoneDefectsofCola1-Jagged1TransgenicMiceTRAPIntratibialinjection:Salineorbreastcancercells(1x104cells)3weeksWTTgBLIimagingmicroCTBonehistologyNormalizedBLI(Log)123WTCol1a-Jag104567***SalineCancerCellsWTSalineCancercellsCol1a-Jag1IncreasedFormationofExperimentalBoneMetastasisinCola1-Jagged1TransgenicMiceJagged1promotesosteolyticbonemetastasisbyengagingNotchsignalinginbonestromalcells.Jag1Ab(15D11)iseffectiveinblockingJagged1-dependentosteolyticbonemetastasis.Jagged1mayalsohaveothertumorpromotingfunction(e.g.chemoresistanceetc),whichcanalsobetargetedby15D11toachievesynergisticeffectwithchemotherapy.Overexpres