Tokyo Medical and Dental UniversityGrants for Excellent Graduate Schools Symposium

February 17th~18th, 2014Tokyo Medical and Dental University

http://www.tmd.ac.jp/cmn/gcoe/index.html

卓越した大学院拠点形成支援補助金国際シンポジウム

Molecular mechanism in bone and tooth,its clinical implication

February 17th~18th, 2014

2014.2.17（Mon）

9：00 − 9：10 PresidentAddress Takashi Ohyama

Openingremark Atsushi Okawa

Session 1 Chairpersons : Bjorn R. Olsen, Atsushi Okawa

9：10 − 10：25 Henry M. Kronenberg Flexibilityintheosteoblastlineage

10：25 − 11：40 Shu Takeda Controlofbonemetabolismviaorgancrosstalk

Session 2 Chairpersons : Benjamin A. Alman, Shohei Kasugai

12：30 − 13：45 Akira Yamaguchi Bonedestructionbyoralcancer

13：45 − 15：00 Hiroshi Takayanagi Forefrontofosteoimmunology

15：00 − 15：30 PosterSession,CoffeeBreak

Session 3 Chairpersons : Ichiro Nishimura, Ichiro Sekiya

15：30 − 16：45 Vicki Rosen Regenerativepotentialofthekneemeniscus

16：45 − 18：00 Hiroshi Asahara Combinatorialapproachtoidentifygeneregulationofinflammatory signalsatRNAlevel

18：30 − Reception

2014.2.18（Tue）

Session 4 Chairpersons : Henry M. Kronenberg, Keiji Moriyama

9：00 − 10：15 Bjorn R. Olsen Craniofacialdysmorphogenesisandinfantilehemangioma– theantraxtoxinreceptorconnection

10：15 − 11：30 Ichiro Sekiya Cartilageandmeniscusregenerationwithsynovialstemcells

Session 5 Chairpersons : Vicki Rosen, Takeshi Muneta

12：20 − 13：35 Benjamin A. Alman Fracturerepairandbeta-catenininaginganddisease: implicationsforanapproachtotherapy

13：35 − 14：50 Ichiro Nishimura OsteoclastasMDSC(myeloid-derivedsuppressorcell)anditsrolein ONJandtumorbonemetastasis

14：50 − 14：55 ClosingRemark Atsushi Okawa

C O N T E N T SHenry M. Kronenberg………………………………………………………… 1Flexibilityintheosteoblastlineage

Shu Takeda …………………………………………………………………… 5Controlofbonemetabolismviaorgancrosstalk

Akira Yamaguchi……………………………………………………………… 9Bonedestructionbyoralcancer

Hiroshi Takayanagi………………………………………………………… 13Forefrontofosteoimmunology

Vicki Rosen ………………………………………………………………… 17Regenerativepotentialofthekneemeniscus

Hiroshi Asahara …………………………………………………………… 21CombinatorialapproachtoidentifygeneregulationofinflammatorysignalsatRNAlevel

Bjorn R. Olsen ……………………………………………………………… 25Craniofacialdysmorphogenesisandinfantilehemangioma–theantraxtoxinreceptorconnection

Ichiro Sekiya………………………………………………………………… 29Cartilageandmeniscusregenerationwithsynovialstemcells

Benjamin A. Alman………………………………………………………… 33Fracturerepairandbeta-catenininaginganddisease:implicationsforanapproachtotherapy

Ichiro Nishimura…………………………………………………………… 37OsteoclastasMDSC(myeloid-derivedsuppressorcell)anditsroleinONJandtumorbonemetastasis

Poster Session……………………………………………………………… 42

1

Flexibility in the osteoblast lineage

Henry M. Kronenberg, M.D.

ProfessorandDirectorEndocrineUnit,MassachusettsGeneralHospitalandHarvardMedicalSchool

Theclassicview,basedprimarilyonstudiesofcellsinculture,isthatmesenchymalprogenitorsinthebonemarrow(called“mesenchymalstemcells”bysomeinvestigators)havethepotentialtodifferentiateintoosteoblasts,chondrocytes,andadipocytes.OsteoblastprecursorsfirstexpressRunx2,thenosterix,andthenothertranscriptionfactorsthatdirectthesecellsfirsttoproliferateandthendifferentiateintomatureosteoblasts.Thefatesofosteoblastsincludetheirdyingbyapoptosisor,instead,becomingosteocytesorinactiveboneliningcells.Invivostudiesusingcrerecombinase,controlledbyvariouspromotersandsometimesrequiringtamoxifenadministrationforactivation(throughactivationofacrerecombinaselinkedtotheligandbindingdomainofamutatedestrogenreceptor)(creERt),haveallowedthemarkingandgeneticmanipulationofcellsintheskeletalsystematvariousstagesofdifferentiationandatspecifictimes.Thesestudieshavedemonstratedunanticipatedflexibilityinthelineageprogramjustoutlined.

Herewewillshowthat,afterosteoblastprecursorsexpressosterixinvivoinmice,removalofβ-cateninexpressionfromthesecellssteersmanyofthesepre-osteoblastsintotheadipocytelineage.Wealsoshowthatmanyosterix-expressingcellsbecomestromalcellsthatsupporthematopoiesis.FurtherweshowthatinactiveboneliningcellscanbereactivatedtoparticipateinboneformationasmatureosteoblastsafterPTHadministration.Finally,weshowthatcollagenII-creERtcanmarkstemcellsthatsubsequentlycanbecomeosteoblasts,chondrocytes,oradipocytesandcontinuetodosoformonthsafteroneadministrationoftamoxifen.Thispromotercanbeactivatedbytamoxifentogeneratemarkedosteoblastsseveralmonthsafterbirth.WhenPTHisgiventothesemice,thenumberofosteoblastprecursorsincreasesintheseveraldaysafterPTHadministration,asindicatedbycountingofthecellsmarkedasexpressingthecollagenIIpromoter.Thus,boththenumberandthefateofearlycellsoftheosteoblastlineagecanbemanipulatedtoservethechangingneedsoftheorganism.

2

Henry M. Kronenberg

CURRICULUM VITAE

Education

1962-1966 B.A.HistoryandLiteratureHarvardCollege1966-1970 M.D.MedicineColumbiaUniversity

Position

1984-1997 Director,EndocrineClinicalLaboratories MassachusettsGeneralHospital1985- MedicalAdministrativeCommitteeMassachusetts GeneralHospital1985-1987 InterimChief,ThyroidUnitMassachusettsGeneral Hospital1986-1989 Director,EndocrineDivisionMassachusetts GeneralHospital1997-2000 Co-Chair,AcademicCouncil,Department ofMedicineMassachusettsGeneralHospital1998-2007 Director,EndocrineDivisionMassachusetts GeneralHospital2007 Chair,CartilageBiologyandPathology ConferenceGordonResearchConferences2009-2010 Director,EndocrineDivisionMassachusetts GeneralHospital2013- Director,EndocrineDivisionMassachusetts GeneralHospital

Award and Honors

2000 VanWyckLectureshipUniversityofNorthCarolina2002 R.L.CreussLecturerMcGillUniversity,Montreal2002 NovartisLecturerCanadianSocietyofEndocrinology& Metabolism2002 5thAnnualJohnG.Haddad,JrDistinguished MemorialLecturer UniversityofPennsylvaniaSchoolofMedicine2003 TransoceanicLecturePrize EuropeanFederationofEndocrineSocieties2003 WilliamF.NeumanAward TheAmericanSocietyforBoneandMineralResearch (ASBMR)2006 LouisV.AvioliMemorialLecturer TheAmericanSocietyforBoneandMineralResearch (ASBMR)2011 Michael&IreneKarlVisitingProfessor WashingtonUniversitySchoolofMedicine2011 2ndIainMacIntyreMemorialLecturer 4thNewYorkSkeletalBiologyandMedicineConference2011 GideonA.RodanAwardforExcellenceinMentoring TheAmericanSocietyforBoneandMineralResearch (ASBMR)2012 TheNinthTorstenN.WieselDistinguishedLecturer HospitalforSpecialSurgery,NewYork,NewYork2012 TriennialInternationalPrize AustrianBoneandMineralSociety2013 JohnBaxterMemorialLecturer

Publications

KobayashiT,LuJ,CobbBS,RoddaSJ,McmahonAP,1.SchipaniE,MerkenschlagerM,Kronenberg,HM.Dicer-dependentpathwaysregulatechondrocyteproliferationanddifferentiation.PNAS2008,105:1949-1954.PMCID:PMC2538863MakKK,KronenbergHM,ChuangPT,MackemS,Yang2.Y.IndianhedgehogsignalsindependentlyofPTHrPtopromotechondrocytehypertrophy.Development2008;135:1947-56.OnoN,NakashimaK,RittlingSR,SchipaniE,HayataT,3.EzuraY,SomaK,DenhardtDT,KronenbergHM,NodaM.Osteopontinnegativelyregulatesparathyroidhormonereceptorsignalinginosteoblasts.JBC2008;283:19400-19409.ProvotS,NachtrabG,ParuchJ,ChenAP,SilvaA,4.KronenbergHM.A-RafandB-RafaredispensablefornormalendochondralbonedevelopmentandPTHrPsuppressesERKactivationinhypertrophicchondrocytes.MolCellBiol.2008;28:344-357.PMCID:PMC2223319WuJY,PurtonLE,RoddaS,ChenM,WeinsteinLS,5.McMahonAP,ScaddenDT,KronenbergHM.OsteoblasticregulationofBlymphopoiesisismediatedbyGsx03B1-dependentsignalingpathways.PNAS2008;105:16976-16981.PMCID:PMC2579363AdamsGB,AlleyIR,ChugUI,ChabnerKT,JeansonNT,6.LoCelsoC,MarstersES,ChenM,WeinsteinLS,LinCP,KronenbergHM,ScaddenDT.HaematopoieticstemcellsdependonGalpha(s)-mediatedsignalingtoengraftbonemarrow.Nature2009;459(7243):103-7.PMCID:PMC2761017BaiXH,WangDW,KongL,ZhangY,LuanY,KobayashiT,7.KronenbergH,YuXP,LiuCJ.ADAMTS-7,adirecttargetofPTHrP,adverselyregulatesendochondralbonegrowthbyassociatingwithandinactivatingGEPgrowthfactor.MolCellBiol2009;29(15)4201-19.PMCID:PMC2715794ConenKL,NishimoriS,ProvotS,KronenbergHM.The8.transcriptionalcofactorLbhregulatesangiogenesisandendochondralboneformationduringfetalbonedevelopment.DevBiol20092009;332(2):348-58.PMCID:PMC2760734MicleaRL,KarperienM,BoschCAJ,vanderHorstG,van9.derValkMA,KobayashiT,KronenbergHM,RawadiG,AkcakayaP,LowikCWGM,FoddeR,WitJ-M,Robanus-MaandagECR.Adenomatouspilyposiscoli-mediatedcontrolofbeta-cateninisessentialforbothchondrogenicandosteogenicdifferentiationofskeletalprecursors.BMCDevelopmentalBiology2009;9:26.PMCID:PMC2678105TerauchiM,LiJ-Y,BediB,BaekK-H,TawfeekH,Galley10.S,GilbertL,NanesMS,ZayzafoonM,GuldbergR,LamarDL,SingerMA,LaneTF,KronenbergHM,WeitzmannMN,PacificiR.TlymphocytesamplifytheanabolicactivityofparathyroidhormonethroughWnt10bsignaling.CellMetab2009;10(3):229-40.PMCID:PMC2761855GuoJ,LiuM,YangD,BouxseinML,SaitoH,GalvinRJS,11.KuhstossSA,ThomasCC,SchipaniE,BaronR,Bringhurst

3

FR,KronenbergHM.SuppressionofWntsignalingbyDKK1attenuatesPTH-mediatedstromalcellresponseandnewboneformation.CellMetabolism2010;11:161-71.PMCID:PMC2819982.KamiyaN,KobayashiT,MochidaY,YuP,Yamauchi12.M,KronenbergH,MishinaY.WntinhibitorsDkk1andSostaredownstreamtargetsofBMPsignalingthroughtheType1Areceptor(BMPR1A)inosteoblasts.JBMR2010:25(2);200-10.PMCID:PMC3153381GuoJ,LiuM,YangD,BouxseinML,ThomasCC,Schipani13.E,BringhurstFR,KronenbergHM.PhospholipaseCsignalingviathePTH/PTHrPreceptorisessentialfornormalbonemodelingandremodeling.Endocrinology2010;151:3502-13.PMCID:PMC2940537MaesC,KobayashiT,SeligMK,TorrekensS,Roth14.SI,MackemS,CarmelietG,KronenbergHM.Osterix-expressingimmatureosteoblastprecursorsinvadegrowthcartilagealongwithbloodvessels.DevelopmentalCell2010;19:329-44.TawfeekH,BediB,LiJ-Y,AdamsJ,KobayashiT,15.WeitzmannMN,KronenbergHM,pacificiR.DisruptionofPTHreceptor1inTcellsprotectsagainstPTH-inducedboneloss.PLoSOne.2010;5(8):e12290.Publishedonline2010August.PMCID:PMC2924900HiraiT,ChaginA,KobayashiT,MackemS,Kronenberg16.H.PTH/PTHrPreceptorsignalingisrequiredformaintenanceofthegrowthplateinpostnatallife.PNAS2011;108(1):191-6.PMCID:PMC3017151MoisanA,RiveraMN,LotinunS,AkhavanfordS,Coffman17.EJ,CookEB,StoykovaS,MukherjeeS,SchoonmakerJA,BurgerA,KimWJ,KronenbergHM,BaronR,HaberDA,BardeesyN.TheWTXtumorsuppressorregulatesmesenchymalprogenitorcellfatespecification.DevCell2011;20(5):583-96.MorishitaM,OnoN,MiyaiK,NakagawaT,HanyuR,18.NagaoM,KamoiratanakulP,NotomiT,RittlingSR,DenhardtDT,KronenbergHM,EzuraY,HayataT,NakamotoT,NodaM.Osteopontindeficiencyenhancesparathyroidhormone/parathyroidhormonerelatedpeptidereceptor(PPR)signaling-inducedalterationintoothformationandodontoblasticmorphology.TissueCell2011;43(3):196-200.OnoN,NakashimaK,SchipaniE,HayataT,EzuraY,19.SomaK,KronenbergHM,NodaM.ConstitutivelyactivePTH/PTHrPreceptorspecificallyexpressedinosteoblastsenhancesboneformationinducedbybonemarrowablation.JCellPysiol2011.Doi:10.1002/jcp22986[Epubaheadofprint].PMID21866553WuJY,AarnisaloP,BastepeM,SinhaP,FulzeleK,Selig20.MK,ChenM,PoultonIJ,PurtonLE,SimsNA,WeinsteinLS,KronenbergHM.Gsαenhancescommitmentofmesenchymalprogenitorstotheosteoblastlineagebutrestrainsosteoblastdifferentiationinmice.JClinInvest2011:121(9):3492-504.PMCID:PMC3163961

Henry M. Kronenberg

HanyuR,WehbiVL,HayataT,MoriyaS,Feinstein21.TN,EzuraY,NagaoM,SaitaY,HemmiH,NotomiT,NakamotoT,SchipaniE,TakedaS,KanekoK,KurosawaH,KarsentyG,KronenbergHM,VilardagaJP,NodaM.Anabolicactionofparathyroidhormoneregulatedbytheβ2-adrenergicreceptor.ProcNatlAcadSciUSA2012;109(19):7433-8.KimSW,PajevicPD,SeligM,BarryKJ,YangJY,Shin22.CS,BaekWY,KimJE,KronenbergHM.IntermittentPTHadministrationconvertsquiescentliningcellstoactiveosteoblasts.JBoneMinerRes.2012may23[Epubaheadofprint].OhishiM,OnoW,OnoN,KhatriR,MarziaM,Baker23.EK,RootSH,WilsonTL,IwamotoY,KronenbergHM,AguilaHL,PurtonLE,SchipaniE.Anovelpopulationofcellsexpressingbothhematopoieticandmesenchymalmarkersispresentinthenormaladultbonemarrowandisaugmentedinamurinemodelofmarrowfibrosis.AmJPathol.2012;180(2):811-8.Epub2011Dec5.OnoN,NakashimaK,SchipaniE,HayataT,EzuraY,24.SomaK,KronenbergHM,NodaM.ConstitutivelyactivePTH/PTHrPreceptorspecificallyexpressedinosteoblastsenhancesboneformationinducedbybonemarrowablation.JCellPhysiol.2012;227(2):408-15.ParkD,SpencerJA,KohBI,KobayashiT,FujisakiJ,25.ClemensTL,LinCP,KronenbergHM,ScaddenDT.EndogenousbonemarrowMSCsaredynamic,fate-restrictedparticipantsinbonemaintenanceandregeneration.CellStemCell2012;10(3):259-72.SongL,LiuM,OnoN,BringhurstFR,KronenbergHM,26.GuoJ.Lostofwnt/β-cateninsignalingcausescellfateshiftofpreosteoblastsfromosteoblaststoadipocytes.JBoneMinerRes.2012;27(11):2344-58.FulzeleK,KrauseDS,PanaroniC,SainiV,BarryKJ,27.LiuX,LotinunS,BaronR,BonewaldL,FengJQ,ChenM,WeinsteinLS,WuJY,KronenbergHM,ScaddenDT,DivietiPajevicP.MyelopoiesisisregulatedbyosteocytesthroughGsα -dependentsignaling.Blood2013;121(6):930-9.GuoJ,SongL,LiuM,SegawaH,MiyamotoK,28.BringhurstFR,KronenbergHM,JüppnerH.ActivationofaNon-cAMP/PKAsignalingpathwaydownstreamofthePTH/PTHrPreceptorisessentialforasustainedhypophosphatemicresponsetoPTHinfusioninmalemice.Endocrinology2013;53(2):487-96.PozziS,FulcinitiM,YanH,ValletS,HomareE,PatelK,29.SantoL,CirsteaD,HideshimaT,SchirtzingeL,KuhstossS,AndersonKC,MunshiN,ScaddenD,KronenbergHM,RajeN.Invivoandinvitroeffectsofanovelanti-Dkk1neutralizingantibodyinmultiplemyeloma.Bone2013;53:487-496.

4

M E M O

5

Itwasbelievedthatcytokinesandhormonesaremainregulatorsofboneremodeling.However,thisviewhasbeenchallenged.Organnetworkhasbeenshowntoplayamajorroleinhomeostasis,recently.Boneisnottheexception.

Clinically,itiswellknownthatheadtraumaacceleratesfracturehealing.Advancesinmoleculargeneticsrevealedthatneuronsandneuropeptides,includingsympatheticnervoussystem,areintimatelyinvolvedinboneremodeling.

Semaphorin3A(Sema3A)isadiffusibleaxonalchemorepellentthatplaysanimportantroleinaxonguidance.PreviousstudieshavedemonstratedthatSema3Aisanosteo-anabolicautocrineand,accordingly,Sema3A-KOmicedevelopalowbonemassduetodecreasedboneformation.However,recently,wedemonstratedthatosteoblast-specificSema3A-KOmicehadnormalbonemass,eventhoughtheexpressionofSema3Ainbonewassubstantiallydecreased.Incontrast,micelackingSema3AinneuronshadlowbonemasssimilartoSema3A-KOmice,indicatingthatneuron-derivedSema3AisresponsiblefortheboneabnormalitiesindependentofthelocaleffectofSema3Ainbone.Indeed,sensoryinnervationsoftrabecularboneweresignificantlydecreasedinneuron-specificSema3A-KO.Moreover,ablatingsensorynervesdecreasedbonemassinwild-typemice,whereasitdidnotdeterioratelowbonemassphenotypeinneuronspecificSema3A-KOmice,furtherindicatingtheessentialroleofsensorynervoussysteminnormalbonehomeostasis.Thus,wedemonstratedthatsensorynervoussystemisalsoacriticalregulatorofboneremodeling.

Inthislecture,Iwouldliketodiscussnovelregulatorsofboneremodeling.

Control of bone metabolism via organ crosstalk

Shu Takeda, M.D., Ph.D.

ProfessorDepartmentofPhysiologyandCellBiology,GraduateSchoolofMedicalandDentalSciences,TokyoMedicalandDentalUniversity

6

Shu Takeda

CURRICULUM VITAE

Education

1992 M.D.UniversityofTokyo,SchoolofMedicine2002 Ph.D.UniversityofTokyo,GraduateSchoolofMedicine

Position

1999 PostdoctralFellow,BaylorCollegeofMedicine, Houston2003 ClinicalFellow,DeptofEndocrinorogyand MetabolismMishukuHospital,Tokyo2004 AssociateProfessor（Junior）,Departmentof OrthopaedicSurgery,21CenterofExcellence Program,GraduateSchool,TokyoMedicaland DentalUniversity2005-2008 AssociateProfessor（Senior）,Departmentof OrthopaedicSurgery,21CenterofExcellence Program,GraduateSchool,TokyoMedicaland DentalUniversity2008 AssociateProfessor,DepartmentofOrthopaedic Surgery,21CenterofExcellenceProgram, GraduateSchool,TokyoMedicalandDental University2009- AssociateProfessor,AssociateProfessor,Section ofNephrology,EndocrinologyandMetabolism, DepartmentofInternalMedicine,Schoolof Medicine,KeioUniversity2013-present Professor,Dept.ofPhysiologyandCellBiology, GraduateSchoolofMedicalandDentalSciences, TokyoMedicalandDentalUniversity2013-present PrincipleInvestigator,CREST,JapanScienceand TechnologyAgency

Award and Honors

1997 PresidentBookAwardofAmericanSocietyforBone andMineralResearch1997 YoungInvestigatorAwardofJapaneseSocietyforBone andMineralMetabolism2000 YoungInvestigatorAwardofAmericanSocietyfor BoneandMineralResearch2007 Correspondingauthorforthepresentation:Most OutstandingAbstractAwardforthe29thAmerican SocietyforBoneandMineralResearchannualmeeting2008 ResearchAwardofJapaneseSocietyforBoneand MineralMetabolism2010 ResearchAwardofJapaneseEndocrineSociety2011 JapanSocietyforthePromotionofSciencePrize2011 BestReviewerof“Bone”2012 AcademicAwardofJapaneseSocietyforBoneand MineralMetabolism2013 MemberoftheAmericanSocietyforClinical Investigation

Publications

Elefteriou,F.,J.D.Ahn,S.Takeda,M.Starbuck,X.Yang,1.X.Liu,H.Kondo,W.G.Richards,T.W.Bannon,M.Noda,K.Clement,C.Vaisse,andG.Karsenty2005LeptinregulationofboneresorptionbythesympatheticnervoussystemandCART.Nature434 :514-20.Fukushima,N.,R.Hanada,H.Teranishi,Y.Fukue,T.2.Tachibana,H.Ishikawa,S.Takeda,Y.Takeuchi,S.Fukumoto,K.Kangawa,K.Nagata,andM.Kojima2005Ghrelindirectlyregulatesboneformation.JBoneMinerRes20 :790-8.Kondo,H.,A.Nifuji,S.Takeda,Y.Ezura,S.R.Rittling,3.D.T.Denhardt,K.Nakashima,G.Karsenty,andM.Noda2005Unloadinginducesosteoblasticcellsuppressionandosteoclasticcellactivationtoleadtobonelossviasympatheticnervoussystem.JBiolChem280 :30192-200.Takeda,S.2005Centralcontrolofboneremodeling.4.BiochemBiophysResCommun328 :697-9.Kamekura,S.,Y.Kawasaki,K.Hoshi,T.Shimoaka,H.5.Chikuda,Z.Maruyama,T.Komori,S.Sato,S.Takeda,G.Karsenty,K.Nakamura,U.I.Chung,andH.Kawaguchi2006Contributionofrunt-relatedtranscriptionfactor2tothepathogenesisofosteoarthritisinmiceafterinductionofkneejointinstability.ArthritisRheum54 :2462-70.Ito,N.,S.Fukumoto,Y.Takeuchi,S.Takeda,H.Suzuki,6.T.Yamashita,andT.Fujita2007Effectofacutechangesofserumphosphateonfibroblastgrowthfactor(FGF)23levelsinhumans.JBoneMinerMetab25 :419-22.Nakamura,T.,Y.Imai,T.Matsumoto,S.Sato,K.Takeuchi,7.K.Igarashi,Y.Harada,Y.Azuma,A.Krust,Y.Yamamoto,H.Nishina,S.Takeda,H.Takayanagi,D.Metzger,J.Kanno,K.Takaoka,T.J.Martin,P.Chambon,andS.Kato2007EstrogenpreventsbonelossviaestrogenreceptoralphaandinductionofFasligandinosteoclasts.Cell130 :811-23.Okuda,N.,S.Takeda,K.Shinomiya,T.Muneta,S.Itoh,M.8.Noda,andY.Asou2007ED-71,anovelvitaminDanalog,promotesboneformationandangiogenesisandinhibitsboneresorptionafterbonemarrowablation.Bone40 :281-92.Sato,S.,R.Hanada,A.Kimura,T.Abe,T.Matsumoto,9.M.Iwasaki,H.Inose,T.Ida,M.Mieda,Y.Takeuchi,S.Fukumoto,T.Fujita,S.Kato,K.Kangawa,M.Kojima,K.Shinomiya,andS.Takeda2007CentralcontrolofboneremodelingbyneuromedinU.NatMed13 :1234-40.Takeda,S.,10. Leptinandbone ,inLeptin ,M.C.Henson,etal.,Editors.2007,Springer.p.1059-1068.Nojiri,Y.,S.Takeda,M.Enomoto,H.Nishitsuji,T.Masuda,11.S.Sotome,andK.Shinomiya2008Co-overexpressionofGDNFandGFRalpha1inducesneuraldifferentiationinneuralprogenitorcellsincomparisontobonemarrowstromalcells.JMedDentSci55 :121-8.Sato,S.,A.Kimura,J.Ozdemir,Y.Asou,M.Miyazaki,T.12.Jinno,K.Ae,X.Liu,M.Osaki,Y.Takeuchi,S.Fukumoto,H.Kawaguchi,H.Haro,K.Shinomiya,G.Karsenty,andS.Takeda2008ThedistinctroleoftheRunxproteins

7

Shu Takeda

inchondrocytedifferentiationandintervertebraldiscdegeneration:Findingsinmurinemodelsandinhumandisease.ArthritisRheum58 :2764-75.Takeda,S.2008Centralcontrolofboneremodelling.13. JNeuroendocrinol20 :802-7.Takeda,S.andP.Ducy,14. RegulationofBoneRemodelingbyCentralandPeripheralNervousSignals ,inPrincipleofBoneBiology ,J.P.Bilezikian,L.G.Raisz,andT.J.Martin,Editors.2008,Elsevier.p.1059-1068.Takeda,S.andG.Karsenty2008Molecularbasesofthe15.sympatheticregulationofbonemass.Bone42 :837-40.Tominaga,H.,S.Maeda,M.Hayashi,S.Takeda,S.Akira,16.S.Komiya,T.Nakamura,H.Akiyama,andT.Imamura2008CCAAT/Enhancer-bindingProtein{beta}PromotesOsteoblastDifferentiationbyEnhancingRunx2ActivitywithATF4.Mol.Biol.Cell19 :5373-5386.Inose,H.,H.Ochi,A.Kimura,K.Fujita,R.Xu,S.Sato,M.17.Iwasaki,S.Sunamura,Y.Takeuchi,S.Fukumoto,K.Saito,T.Nakamura,H.Siomi,H.Ito,Y.Arai,K.-i.Shinomiya,andS.Takeda2009AmicroRNAregulatorymechanismofosteoblastdifferentiation.ProcNatlAcadSciUSA106 :20794-20799.Suzuki,D.,A.Yamada,T.Amano,R.Yasuhara,A.Kimura,18.M.Sakahara,N.Tsumaki,S.Takeda,M.Tamura,M.Nakamura,N.Wada,T.Nohno,T.Shiroishi,A.Aiba,andR.Kamijo2009EssentialmesenchymalroleofsmallGTPaseRac1ininterdigitalprogrammedcelldeathduringlimbdevelopment.DevBiol335 :396-406.Takeda,S.2009Osteoporosis:Aneuroskeletaldisease?19. IntJBiochemCellBiol41 :455-9.Itoh,T.,S.Takeda,andY.Akao2010MicroRNA-20820.modulatesBMP-2-stimulatedmousepreosteoblastdifferentiationbydirectlytargetingV-etserythroblastosisvirusE26oncogenehomolog1.JBiolChem285 :27745-52.Kimura,A.,H.Inose,F.Yano,K.Fujita,T.Ikeda,S.Sato,21.M.Iwasaki,T.Jinno,K.Ae,S.Fukumoto,Y.Takeuchi,H.Itoh,T.Imamura,H.Kawaguchi,U.I.Chung,J.F.Martin,S.Iseki,K.Shinomiya,andS.Takeda2010Runx1andRunx2cooperateduringsternalmorphogenesis.Development137 :1159-67.Nishikawa,K.,T.Nakashima,S.Takeda,M.Isogai,M.22.Hamada,A.Kimura,T.Kodama,A.Yamaguchi,M.J.Owen,S.Takahashi,andH.Takayanagi2010Mafpromotesosteoblastdifferentiationinmicebymediatingtheage-relatedswitchinmesenchymalcelldifferentiation.JClinInvest120 :3455-65.Sato,T.,D.Chida,T.Iwaya,M.Usui,M.Hatori,T.Abe,23.S.Takeda,andT.Yoda2010Non-neuronalregulationandrepertoireofcholinergicreceptorsinorgans.BioMolecularConceptsinpress .Takeda,S.,24. SkeletalRegulationofEnergyMetabolism,inBoneandDevelopmentinTopicinBoneBiology ,F.Bronner,H.Roach,andM.Farach-Carson,Editors.2010,Springer.p.267-278.

Tanaka,M.,T.Suganami,S.Sugita,Y.Shimoda,M.25.Kasahara,S.Aoe,M.Takeya,S.Takeda,Y.Kamei,andY.Ogawa2010Roleofcentralleptinsignalinginrenalmacrophageinfiltration.EndocrJ57 :61-72.Meguro,S.,M.Tomita,T.Katsuki,K.Kato,H.Oh,A.26.Ainai,R.Ito,S.Takeda,T.Kawai,Y.Atsumi,H.Itoh,andH.Hasegawa2011Plasma25-hydroxyvitamindisindependentlyassociatedwithhemoglobinconcentrationinmalesubjectswithtype2diabetesmellitus.IntJEndocrinol2011 :362981.Nagao,M.,T.N.Feinstein,Y.Ezura,T.Hayata,T.Notomi,27.Y.Saita,R.Hanyu,H.Hemmi,Y.Izu,S.Takeda,K.Wang,S.Rittling,T.Nakamoto,K.Kaneko,H.Kurosawa,G.Karsenty,D.T.Denhardt,J.-P.Vilardaga,andM.Noda2011Sympatheticcontrolofbonemassregulatedbyosteopontin.ProceedingsoftheNationalAcademyofSciences .Ogata,N.,Y.Shinoda,N.Wettschureck,S.Offermanns,28.S.Takeda,K.Nakamura,G.V.Segre,U.-i.Chung,andH.Kawaguchi2011GαqSignalinOsteoblastsIsInhibitorytotheOsteoanabolicActionofParathyroidHormone.JournalofBiologicalChemistry286 :13733-13740.Hinoi,E.,Ochi,H.,Takarada,T.,Nakatani,E.,Iezaki,T.,29.Nakajima,H.,Fujita,H.,Takahata,Y.,Hidano,S.,Kobayashi,T.,Takeda,S.,andYoneda,Y.Positiveregulationofosteoclasticdifferentiationbygrowthdifferentiationfactor15upregulatedinosteocyticcellsunderhypoxia.JBoneMinerRes27 :938-49,2012Hanyu,R.,Wehbi,V.L.,Hayata,T.,Moriya,S.,Feinstein,30.T.N.,Ezura,Y.,Nagao,M.,Saita,Y.,Hemmi,H.,Notomi,T.,Nakamoto,T.,Schipani,E.,Takeda,S.,Kaneko,K.,Kurosawa,H.,Karsenty,G.,Kronenberg,H.M.,Vilardaga,J.-P.,andNoda,M.Anabolicactionofparathyroidhormoneregulatedbytheβ2-adrenergicreceptor.ProceedingsoftheNationalAcademyofSciences109 :7433-7438,2012Fujita,K.,Iwasaki,M.,Ochi,H.,Fukuda,T.,Ma,C.,31.Miyamoto,T.,Takitani,K.,Negishi-Koga,T.,Sunamura,S.,Kodama,T.,Takayanagi,H.,Tamai,H.,Kato,S.,Arai,H.,Shinomiya,K.,Itoh,H.,Okawa,A.,andTakeda,S.VitaminEdecreasesbonemassbystimulatingosteoclastfusion.NatMed18 :589-594,2012Iwasaki,M.,Piao,J.,Kimura,A.,Sato,S.,Inose,H.,Ochi,32.H.,Asou,Y.,Shinomiya,K.,Okawa,A.,andTakeda,S.Runx2haploinsufficiencyamelioratesthedevelopmentofOssificationofthePosteriorLongitudinalLigament.PLoSONE.7 :e43372,2012Fukuda,T.,Takeda,S.,Xu,R.,Ochi,H.,Sunamura,S.,Sato,33.T.,Shibata,S.,Yoshida,Y.,Gu,Z.,Kimura,A.,Ma,C.,Xu,C.,Bando,W.,Fujita,K.,Shinomiya,K.,Hirai,T.,Asou,Y.,Enomoto,M.,Okano,H.,Okawa,A.,andItoh,H.Sema3Aregulatesbone-massaccrualthroughsensoryinnervations.Nature2013

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Oralsquamouscellcarcinoma(OSCC)frequentlyinvadesthejaws,andthisinvasionisassociatedwithaworseprognosis.Themechanismunderlyingtheboneinvasionremainspoorlyunderstood.Wehistopathologicallyinvestigatedmandibularinvasionpatternsin97casesofprimaryOSCC,andshowedthatinallcasesvaryingamountsoffibrousconnectivetissuesintervenedbetweenthetumorcellsandthebone.ImmunohistochemistryrevealedRANKLexpressioninthefibroblasticcellsthatwereadjacenttotheosteoclastsintheareaofboneresorption.Theseresultssuggestthatthefibrousstromaisinvolvedinosteoclasticboneresorption.Toexplorethemolecularmechanism,weconductedmicroarrayanalysesusing43humanOSCCspecimens,andrevealedthatmanyofspecimensoverexpressedPTHrPmRNA,butafewoverexpressedIL-6 mRNA.ImmunohistochemicalanalysisrevealedthatIL-6wasexpressednotonlyincancercellsbutalsoinfibroblastsattumor-boneinterface.Conditionedmedia(CM)derivedfromthecultureoforalcancercelllinesstimulatedRanklexpressioninstromalcellsandosteoclastformation.AntibodiesagainstbothhumanPTHrPandmouseIL-6receptorsuppressedRankl inST2cellsandosteoclastformationinducedbyCM.XenograftsofOSCCcellsontotheperiostealregionoftheparietalboneinathymicmicepresentedsimilarhistologyandexpressionprofilesofRANKLandIL-6asobservedinbone-invasivehumanOSCCspecimens.TheseresultsindicatethatOSCCprovidesasuitablemicroenvironmentforosteoclastformationnotonlybyproducingIL-6andPTHrPbutalsobystimulatingstromalcellstosynthesizeIL-6.RecentreportsindicatedthesynthesisofRANKLbyOSCCcellsaswellasthetumorstromalcells.Indeed,HSC3andHO-1-N-1,humanOSCCcelllines,expressedRANKLandstimulatedRankl expressioninUAMS-32murineosteoblasticcellline.WediscriminatedtherolesofRANKLsynthesizedbystromalcellsandcancercellsincancer-associatedboneresorptionbyusingspecies-specificRANKLantibodiesagainstmurineRANKLandhumanRANKL,respectively.OsteoclastogenesisinducedbytheconditionedmediumofHSC3andHO-1-N-1cellsinaco-cultureofmurinebonemarrowcellsandUAMS-32cellswasinhibitedbytheadditionofantibodiesagainsteithermouseorhumanRANKL.HSC3-inducedbonedestructionwasgreatlyinhibitedbytheadministrationofanti-mouseRANKLantibodyinaxenograftmodel.HO-1-N-1-inducedbonedestructionwasinhibitedbytheadministrationofeitheranti-mouseoranti-humanRANKLantibody.BonedestructioninducedbythetransplantationofhumanRANKL-overexpressingcells(HSC3-R2)wasgreatlyinhibitedbytheinjectionofanti-humanRANKLantibody.ThepresentstudyrevealedthatRANKLproducedbybothstromalandcancercellsisinvolvedinoralcancer-inducedosteoclasticboneresorption.Theseresultsprovideimportantinformationforunderstandingthecellularandmolecularbasisofcancer-associatedbonedestructionandthemechanismofactionunderlyingRANKLantibody(denosumab)therapy.ToexplorethemoleculesthatsynthesizedbyOSCCcellsandstimulateosteoclasticboneresorption,weestablishedtwoclonalcelllines,HSC3-C13andHSC3-C17,fromthematernaloralcancercellline,HSC3.TheconditionedmediumfromHSC3-C13cellsshowedthehighestinductionofRankl expressioninthemousestromalcelllinesST2andUAMS-32ascomparedtothatinmaternalHSC3cellsandHSC3-C17cells,whichshowedsimilaractivity.TheconditionedmediumfromHSC3-C13cellssignificantlyincreasedthenumberofosteoclastsinaco-culturewithmousebonemarrowcellsandUAMS-32cells.XenografttumorsgeneratedfromtheseclonalcelllinesintotheperiostealregionoftheparietalboneinathymicmiceshowedthatHSC3-C13cellscausedextensivebonedestructionandasignificantincreaseinosteoclastnumbersascomparedtoHSC3-C17cells.GeneexpressionwascomparedbetweenHSC3-C13andHSC3-C17cellsbyusingmicroarrayanalysis,whichshowedthatCXCL2genewashighlyexpressedinHSC3-C13cellsascomparedtoHSC3-C17cells.ImmunohistochemicalstainingrevealedthelocalizationofCXCL2inhumanOSS.TheincreaseinosteoclastnumbersinducedbytheHSC3-C13-conditionedmediumwasdose-dependentlyinhibitedbyadditionofanti-humanCXCL2-neutralizingantibodyinaco-culturesystem.RecombinantCXCL2increasedtheexpressionofRankl inUAMS-32cells.TheseresultsindicatethatCXCL2isinvolvedinbonedestructioninducedbyoralcancer.ThisisthefirstreportshowingtheroleofCXCL2incancer-associatedbonedestruction.

Bone destruction by oral cancer

Akira Yamaguchi, D.D.S., Ph.D.

ProfessorDepartmentofOralPathology,GraduateSchoolofMedicalandDentalSciences,TokyoMedicalandDentalUniversity

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Akira Yamaguchi

CURRICULUM VITAE

Education

1974 D.D.S.TokyoDentalCollege1980 Ph.D.TokyoMedicalandDentalUniversity

Position

1980-1988 AssistantProfessor,DepartmentofOralPathology, SchoolofDentistry,ShowaUniversity1985-1987 VisitingAssistantProfessor,SchoolofDentistry, WashingtonUniversity,Si.Loius,MO,USA1987-1988 VisitingAssistantProfessor,OrthopedicSurgery, SchoolofMedicine,St.LouisUniversity1988-1998 AssociateProfessor,DepartmentofOralPathology, SchoolofDentistry,ShowaUniversity1998-2004 ProfessorandChair,DepartmentofOral Pathology,UniversitySchoolofDentistry2004- ProfessorandChair,DepartmentofOralPathology GraduateSchoolofMedicalandDentalSciences TokyoMedicalandDentalUniversity2011- Councilmember,ScienceCouncilofJapan

Award and Honors

1993 TheJapaneseSocietyforBoneandMineralResearch: AcademicAward1996 TheJapaneseSocietyofPathology:PathologyResearch Award2005 TheJapanesePathologyAward

Publications

YamaguchiA,KatagiriT,IkedaT,WozneyJM,1.RosenV,WangEA,KahnAJ,SudaT,YoshikiS:Recombinanthumanbonemorphogeneticprotein-2stimulatesosteoblasticmaturationandinhibitsmyogenicdifferentiationinvitro.JCellBiol 111:681-687,1991KatagiriT,YamaguchiA,KomakiM,AbeE,TakahashiN,2.IkedaT,RosenV,WozneyJM,Fujisawa-SeharaA,SudaT:Bonemorphogeneticprotein-2convertsthedifferentiationpathwayofC2C12myoblastsintotheosteoblastlineage.JCellBiol 127:1755-1766,1994IshizuyaT,YokoseS,HoriM,NodaT,SudaT,Yoshiki3.S,YamaguchiA:Parathyroidhormoneexertsdisparateeffectsonosteoblastdifferentiationdependingonexposuretimeinratosteobllasticcells.JClinInvest 99:2961-2970,1997KomoriT,YagiH,NomuraS,YamaguchiA,SasakiK,4.DeguchiK,ShimuzuY,BronsonRT,GaoY-H,InadaM,SatoM,OkamotoR,TominagaK,KitamuraY,YoshikiS,KishimotoT:TargeteddisruptionofCbfa1resultsincompletelackofboneformationowingtothematurationarrestofosteoblasts.Cell 89:755-764,1997KintoN,IwamotoM,Enomoto-IwamotoM,NojiS,Ohuchi5.H,YoshidaH,KataokaH,WadaY,GaoY-H,TkahashiHE,YoshikiS,YamaguchiA:FibroblastsexpressingSonichedgehoginduceosteoblastdifferentiationandectopicboneformation.FEBSLetters404:319-323,1997YamaguchiA,KomoriT,SudaT:RegulationofOsteoblast6.DifferentiationMediatedbyBMPs,HedgehogsandCbfa1.EndocrineReviews21:393-411,2000YuasaT,KataokaH,KintoN,IwamotoM,Enomoto-7.IwamotoM,IemuraS,UenoN,ShibataY,KurosawaH,YamaguchiA:SonichedgehogisinvolvedinosteoblastdifferentiationbycooperatingwithBMP-2.JCell.Physiol 193:225-232,2002NobtaM,TsukazakiT,ShibataY,ChangX,MoriishiT,8.SakanoS,ShindoH,YamaguchiA:CriticalregulationofBMP-inducedosteoblasticdifferentiationbyDelta1/Jagged1-activatedNotch1signaling.JBiolChem280:15842-15848,2005LiuT,GaoY,SakamotoK,MinamizatoT,FurukawaK,9.TsukazakiT,ShibataY,BesshoK,KomoriT,YamaguchiA:BMP-2promotesdifferentiationofosteoblastsandchondroblastsinRunx2-deficientcelllines.JCellPhysiol211:728-735,2007MinamizatoT,SakamotoK,LiuT,KokuboH,KatsubeK,10.PerbalB,NakamuraS,YamaguchiA:CCN3/NOVinhibitsBMP-2-inducedosteoblastdifferentiationbyinteractingwithBMPandNotchsignalingpathways.BiochemBiophResCo354:567-573,2007IshikuroM,SakamotoK,KayamoriK,IzumoT,Yamaguchi11.A:Roleoffibrousstromainboneinvasionbygingivalsquamouscellcarcinomas.BONE43:621-627,2008KayamoriK,SakamotoK,NakashimaT,TakayanagiT,12.

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MoritaK,OmuraK,NguyenST,MikiY,IimuraT,HimenoA,AkashiT,Yamada-OkabeH,OgataE,YamaguchiA:RolesofIL-6andPTHrPinosteoclastformationassociatedwithoralcancers:ThesignificanceofIL-6synthesizedbystromalcellsinresponsetocancercells.AmerJPathol 176,968-980,2010SakamotoK,AragakiT,KawachiH,KatsubeK,MikiY,13.TakizawaT,OmuraK,MoritaK,OkadaN,YamaguchiA:Down-regulationofkeratin4andkeratin13expressioninoralsquamouscellcarcinomaandepithelialdysplasia:aclueforhistopathogenesis.Histopathol 58:531-542,2011OueE,LeeJW,SakamotoK,IimuraT,AokiK,KayamoriK,14.MichiY,YamashiroM,YamaguchiA:CXCL2synthesizedbyoralsquamouscellcarcinomaisinvolvedincancer-associatedbonedestruction.BiochemBiophResCo424:456-461,2012WatanabeT,TamamuraY,HoshinoA,MakinoY,15.NishimuraR,KamiokaH,YonedaT,AmagasaT,YamaguchiA,IimuraT:IncreasingparticipationofSclerostininpostnatalbonedevelopmentrevealedbythree-dimensionalimmunofluorescencemorphometry.BONE51:447-458,2012SakamotoK,FujiiT,KawachiH,MikiY,OmuraK,Morita16.K,KayamoriK,KhanomR,KatsubeK,YamaguchiA:ReductionofNOTCH1expressionpertainstomaturationabnormalitiesofkeratinocytesinsquamousneoplasms.LabInvest 92:688-702:2012MatsushitaY,SakamotoK,TamamuraY,ShibataY,17.MinamizatoT,KiharaT,ItoM,KatsubeK,HiraokaS,KosekiH,HaradaK,YamaguchiA:CCN3proteinparticipatesinboneregenerationasaninhibitoryfactor.JBiolChem288:19973-19985,2013SatoK,LeeJW,SakamotoK,IimuraT,KayamoriK,18.YasudaH,ShindohM,ItoM,OmuraK,YamaguchiA:RANKLsynthesizedbybothstromalcellsandcancercellsplaysacrucialroleinosteoclasticboneresorptioninducedbyoralcancer.AmerJPathol 182:1890-1899,2013

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Thebonemarrowisoneoftheprimarylymphoidorgansthatharbortheimmunecells.Underthesemicroenvironments,bonecellsinteractnotonlywithotherbonecellsbutalsowithimmunecellsincludinghematopoieticstemcells.Thus,allthecellsinthebonemarrowarecommunicatingwitheachothertomaintainbothskeletalandimmunesystems(1,2).However,themoleculesthatmediatethecommunicationamongcellsinthebonehavebeenpoorlyidentified.Wehavetriedtoexplorethemolecularbasisforcellularcommunicationinthebonemarrowandfoundthatosteocyte-derivedRANKLplaysacrucialroleinadultosteoclastogenesis(3),osteoclast-derivedSema4Dinhibitsboneformation(4)andosteoblast-derivedSema3Afunctionsasapotentosteoprotectivecytokineinhibitingosteoclastogenesisandpromotingosteoblastogenesis(5).Assemaphorins,majoraxonguidancemolecules,haveemergedasbonecellcommunicationfactors,wearefacinganewstageofboneresearchincludingthebone,immuneandneuralsystems.WehavelongworkedontheinteractionbetweenTcellsandosteoclasts,andrecentlyfoundtheoriginofosteoclastogenicTh17cellstobeFoxp3positiveTregs(6).TherecentadvancesinosteoimmunologyfieldincludingtheconversionofTregtopathogenicTcellswillbediscussed.

1. TakayanagiH.NatRevImmunol7,292-304(2007),2. TakayanagiH.NatRevRheumatol5(12),667-76(2009),3. NakashimaT.etal.NatMed17,1231-34(2011),4. Negishi-KogaT.etal.NatMed17,1473-1480(2011),5. HayashiM.etal.Nature485,69-74(2012)6. KomatsuN.etal.NatMed,inpress

Forefront of osteoimmunology

Hiroshi Takayanagi, M.D., Ph.D.

ProfessorDepartmentofImmunology,GraduateSchoolofMedicineandFacultyofMedicine,TheUniversityofTokyo

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Hiroshi Takayanagi

CURRICULUM VITAE

Education

1990 M.D.TheUniversityofTokyo,Tokyo,Japan2001 Ph.D.(Medicine)TheUniversityofTokyo,Tokyo,Japan

Award and Honors

2002 AmershamBiosciencesandSciencePrizeforYoung Scientists2002 NovartisJapanRheumatologyPrize2004 JapanCollegeofRheumatologyAward2004 FullerAlbrightAward,ASBMR2005 JapanSocietyforthePromotionofSciencePrize2005 JapanAcademyMedal2006 InternationalResearchPrize,AustrianSocietyforBone andMineralResearchandLudwigBoltzmannInstitute ofOsteology2008 AcademicAwardoftheMochidaMemorialFoundation2009 InouePrizeforScience2009 JSBMRDistinguishedScientistAward2011 IBMS-BONEHerbertA.FleischAward

Publications

TakayanagiHetal.1. Nature408,600-605(2000)TakayanagiHetal.2. Nature416,744-749(2002)TakayanagiHetal.3. DevCell 3,889-901(2002)KogaTetal.4. Nature428,758-763(2004)TakayanagiH.5. NatRevImmunol 7,292-304(2007)AsagiriMetal.6. Science319,624-627(2008)ShinoharaMetal.7. Cell 132,794-806(2008)Takayanagi,H8. NatRevRheumatol 5(12),667-76(2009)OkamotoKetal.9. Nature464,1381-5(2010)NakashimaTetal.10. NatMed17,1231-34(2011)Negishi-Koga,T.,etal.11. NatMed17,1473-1480(2011)Hayashi,M.,etal.12. Nature485,69-74(2012)TakayanagiHetal.13. CellMetab17(2):249-60.(2013)

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Osteoarthritis(OA)affectsover200millionpeopleworldwide.JointsmostfrequentlyaffectedbyOAarethehands,hips,spineandknees,wherebreakdownandsubsequentlossofarticularcartilageresultsinpain,stiffnessandswellingandleadstodecreasedjointfunctionandlossofjointmobility.ThekneejointisparticularlysusceptibletoOAandmeniscalinjuriesarethemostcommonkneeinjury.Treatmentofmeniscalinjurieshasevolveddramaticallyinthepast30yearsinresponsetoanincreasedunderstandingoftheroleperformedbythemeniscuswithinthekneejoint,andtoday,surgicalproceduresareaimedatrepairingorreplacingdamagedmenisci,dependingonthesizeoftheinjuryanditslocation.Unfortunately,damagedmeniscaltissuerarelyregainsthestructuralintegrityandmechanicalstrengthofnormalmeniscus,andrecentdataindicatethatsurgicalrepairofmeniscaltearscannotreliablypreventtheprogressionofdegenerativechangesandclinicalsymptomsthatpresagethedevelopmentofkneeOA.Particularlystrikingisthelackofoptionsforreversingtheprogressivedebilitationofthekneeinyoung,healthyindividuals.

Evidencefromrecentstudieslinksthesignalingpathwaysdirectingembryonicdevelopmentwiththosethatmediateadulttissueregeneration,wheretheyinfluencethemigration,proliferationanddifferentiationoflocallyresidingtissue-specificstem/progenitorcells.Inthisway,thespecificityofthereparativeresponsegeneratedbyatissueisdependentontheactivationoftheresidenttissue-specificstem/progenitorcellpopulationandnotonthesignalsthataffecttheirbehavior.Foradultmusculoskeletaltissues,resident-specificstem/progenitorcellshavebeenidentifiedandcharacterizedinmuscle,tendonandligament,articularcartilageandbone.Asagroup,thesetissue-specificstemcellspossesssomeofthecharacteristicsofmesenchymalstemcells(MSC)butmoreimportantly,exhibitcharacteristictissue-specificidentities.Basedonthehypothesisthatcellsnecessaryformeniscalrepairarethosethatexpressacharacteristicgenesignaturethatisdefinedduringmeniscalmorphogenesiswecombinedlaser-capturemicro-dissection(LCM)ofembryonicjointtissueswiththeproductionoftissue-specificlibrariesandcomparativegenearrayanalysistouncoverthegenesignatureofmeniscusformingcellsandthesignalingpathwaysthatregulateexpressionofthesegenes.Currenteffortsareaimedatusinginformationgainedfromthestudyofmeniscusdevelopmentinmicetoquerythereparativepotentialofcellscollectedfromadultmouseandhumanmeniscus.

Regenerative potential of the knee meniscus

Vicki Rosen, Ph.D.

ProfessorDepartmentofDevelopmentalBiology,HarvardSchoolofDentalMedicine

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Vicki Rosen

CURRICULUM VITAE

Education

1975 B.S.,Biology,StateUniversityofNewYorkatStony Brook1981 Ph.D.,CellBiologyandPhysiology,Universityof Connecticut

Position

1984-1988 StaffScientistII,BMPDiscoveryResearch, GeneticsInstitute1986-1988 PrincipalScientist,BMPDiscoveryResearch, GeneticsInstitute1988-1998 SeniorScientist,BMPDiscoveryResearch, GeneticsInstitute1998-2001 Director,TissueGrowthandRepair,Genetics Institute2001 DistinguishedResearchScientist,Genetics Institute/WyethResearch2001-2005 SeniorMemberoftheStaff,TheForsythInstitute2001-present ProfessorofDevelopmentalBiology, HarvardSchoolofDentalMedicine2005-present Chair,DepartmentofDevelopmentalBiology, HarvardSchoolofDentalMedicine

Award and Honors

2001 HonoraryMA,HarvardUniversity2008 KappaDeltaAward,co-recipient,OrthopedicResearch Society2010 MarshallUristAwardinTissueRepair,Orthopedic ResearchSociety2010 BasicScienceChair,2010ASBMRMeeting2012 Harvard-AustraliaFoundationFellowship2012 RaineMedicalResearchFoundationMedal

Publications

LanchoneyTF,OlmsteadEA,ShoreEM,GannonFA,1.RosenV,ZasloffMA,andKaplanFS.Characterizationofbonemorphogeneticprotein4receptorinfibrodysplasiaossificansprogressiva.ClinOrthopRelRes1998;346:38-45.ChenD,JiX,HarrisMA,FengJQ,KarsentyG,Celeste2.AJ,RosenV,MundyGR,andHarrisSE..Differentialrolesforbonemorphogeneticprotein(BMP)receptortypeIBandIAindifferentiationandspecificationofmesenchymalprecursorcellstoosteoblastandadipocytelineages.JCellBiol 1998;142:295-305.FuruyaK,NifujiA,RosenV,andNodaM.Effectsof3.GDF7/BMP12onproliferationandalkalinephosphataseexpressioninratosteoblasticosteosarcomaROS17/2.8cells.JCellBiochem1999;72:177-180.LeeM-H,JaavedA,KimH-J,ShinH-I,GutierrezS,Choi4.J-Y,RosenV,SteinJL,vanWijnenAJ,SteinGS,LianJB,andRyooH-M.TransientupregulationofCBFA1inresponsetobonemorphogeneticprotein-2andtransforminggrowthfactorß1inC2C12myogeniccellscoincideswithsupressionofthemyogenicphenotypebutisnotsufficientforosteoblastdifferentiation.JCellBiochem1999;73:114-125.NeuhausH,RosenV,andThiesRS.Heartspecific5.expressionofmouseBMP-10,anovelmemberoftheTGF-ßsuperfamily.MechDev1999;80:181-184.GamerLW,WolfmanNM,CelesteAJ,HattersleyG,6.HewickR,andRosenV.AnovelBMPexpressedindevelopingmouselimb,spinalcord,andtailbudisapotentmesoderminducerinXenopusembryos.DevBiol 1999;208:222-232.ValcourtU,RonziereM-C,WinklerP,RosenV,Herbage7.D,andMallein-GerinF.1999.Differenteffectsofbonemorphogeneticproteins2,4,12,and13ontheexpressionofcartilageandbonemarkersintheMC615chondrocytecellline.ExpCellRes251:264-274.LadherRHK,ChurchVL,AllenS,RobsonL,Abdelfattah8.A,BrownNA,HattersleyG,RosenV,LuytenFP,DaleL,andFrancis-WestPH.CloningandexpressionoftheWntantagonistsSfrp-2andFrzbduringchickdevelopment.DevBiol 2000;218:183-198.WestonAD,RosenV,andUnderhillTM.Regulationof9.skeletalprogenitordifferentiationbytheBMPandretinoidsignalingpathways.JCellBiol 2000;148:679-690.YiSE,DaluiskiA,PedersonR,RosenV,LyonsKM.The10.typeIBMPRIBisrequiredforchondrogenesisinthemouselimb.Development2000;127(3):621-30.DailuskiA,EngstrandT,BahamondeM,TompkinsonK,11.GamerLW,RosenV,andLyonsKM.BMP3isanegativeregulatorofbonedensity.NatureGenet2001;27:84-88.GamerLW,CoxKA,SmallC,andRosenV.Gdf11isa12.negativeregulatorofchondrogenesisandmyogenesisinthedevelopingchicklimb.DevBiol 2001;229:407-420.BanerjeeC,JavedA,ChoiJ-Y,GreenJ,RosenV,van13.

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Vicki Rosen

WijnenAJ,SteinJL,LianJB,andSteinGS.DifferentialregulationofthetwoprincipalRunX/Cbfa1N-terminalisoformsinresponsetoBMP-2duringdevelopmentoftheosteoblastphenotype.Endocrinol 2001;142:4026-4039.SchweitzerR,ChyungJH,MurtaughLC,BrentAE,14.RosenV,OlsenEN,LassarA,andTabinCJ.AnalysisofthetendoncellfateusingScleraxis,aspecificmarkerfortendonsandligaments.Develop2001;128:3855-3886.ThomopoulosS,HattersleyG,RosenV,MertensM,Galatz15.L,WilliamsGR,andSoslowskyLJ.Thelocalizedexpressionofextracellularmatrixcomponentsinhealingtendoninsertionsites:aninsituhybridizationstudy.JOrthopRes2002;20:454-463.Gamer,LW,Nove,J,andRosen,V.ReturnoftheChalones.16.DevCell 2003;4(3):143-144.Gamer,LW,Nove,J,Levin,M,andRosen,V.BMP-3isa17.novelinhibitorofBMP-4andactivininXenopus.DevBiol 2005;285:156-168.ChenH,YongW,RenS,ShenW,HeY,CoxKA,ZhuW,18.LiW,SoonpaaM,PayneRM,FrancoD,FieldLJ,RosenV,WangY,andShouW.OverexpressionofBMP10inmyocardiumdisruptscardiacpostnatalhypertrophicgrowth.JBiolChem2006;281:27481-27491.BandyopadhyayA,TsujiK,HarfeBD,RosenV,andTabin19.CJ.GeneticanalysisoftherolesofBMP2,BMP4andBMP7inlimbpatterningandskeletogenesis.PLOSGenetics 2006;2:e216TsujiK,BandyopadhyayA,HarfeBD,CoxKA,Kakar20.S,GerstenfeldL,EinhornT,TabinCJ,andRosen,V.BMP2activity,althoughdispensableforboneformation,isrequiredfortheinitiationoffracturehealing.NatGenet .2006;38:1424-1429.TsujiK,BandyopadhyayA,HarfeBD,CoxKA,TabinCJ,21.andRosenV.BMP4isdispensableforskeletogenesisandfracturehealinginthelimb.JBoneJointSurg .2008;90(Suppl1):14-18.GamerLW,HoV,CocK,andRosenV.Expressionand22.functionofBMP3duringchicklimbdevelopment.DevDyn2008;237:1691-1698.GamerLW,CoxK,CarloJM,andRosenV.23.OverexpressionofBMP3inthedevelopingskeletonaltersendochondralboneformationresultinginspontaneousribfractures.DevDyn .2009;238:2374-2381.AlamN,St-ArnaudR,LauzierD,RosenV,andHamdyRC.24.AreendogenousBMPsnecessaryforbonehealingduringdistractionosteogenesis?ClinOrthopRelatRes .2009;467:3190-3198.TsujiK,CoxK,GamerL,GrafD,EconomidesA,and25.RosenV.ConditionaldeletionofBMP7fromthelimbskeletondoesnotaffectboneformationorfracturerepair.JOrthopRes .2010;28:384-389.StankovicKM,AdachiO,TsujiK,KristiansenAG,Adams26.JC,RosenV,andMcKennaMJ.Differencesingeneexpressionbetweentheoticcapsuleandotherbones.Hear

Res ,2010;265:83-9.PMID:20146935LoweryJW,PazinD,IntiniG,KobabuS,ChappiusV,27.CapeloLandRosenV.TheroleofBMP2signalingintheskeleton.CritrevEukaryotGeneExpr2011;21:177-185.PMID:22077155LoweryJWandRosenV.Allele-specificRNAinterference28.inFOPsilencingtheFOPgene.GeneTher2012;19:701-2.PMID:22130446GamerLW,TsujiK,CoxK,CapeloLP,LoweryJ,BeppuH,29.RosenV.“BMPR-IIisdispensableforformationofthelimbskeleton”Genesis .2011Sep:49(9):719-24.PMID:2153880KokabuS,GamerL,CoxK,LoweryJW,TsujiK,30.EconomidesA,KatagiriT,RosenV.“BMP3suppressesosteoblastdifferentiationofbonemarrowstromalcellsviainteractionwithAcvr2b.”JMolEndocrinol .2012Jan;26(1):87-94.PMID:22074949ChappuisV,GamerL,CoxK,LoweryJW,BosshardtDD31.andRosenV.PeriostealBMP2activitydrivesbonegrafthealing.Bone2012;51:800-809.PMID:22846673Pazin,DE,GamerLW,CoxKA,RosenV.“Molecular32.profilingofsynovialjoints:useamicroarrayanalysistoidentifyfactorsthatdirectthedevelopmentofthekneeandelbow.”DevDyn2012;241(11):1816-26.PMID:22972626LoweryJW,LavigneAW,KokabuS,andRosenV.33.ComparativegenomicsidentifiesthemouseBmp3promoterandanupstreamevolutionaryconservedregion(ECR)inmammals.PLoSOne2013;8:e57840PMID:23451274WhitmanM,RosenV,BrivanlouAH,GroppeJC,Sebald34.WandMuellerT.Regardingthemechanismofactionofaproposedpeptideagonistofthebonemorphogeneticproteinreceptoractivin-likekinase3.NatureMed2013;19:809-10.PMID:23836213KokabuS,NguyenT,OhteS,SatoT,KatagiriT,Yoda35.TandRosenV.TLE3,transducing-likeenhancerofsplit3,suppressesosteoblastdifferentiationofbonemarrowstromalcells.BiochemBiophysResCommun2013;438:205-10.SchminkeB,MuhammadH,BodeC,SadowskiB,Gerter36.R,GersdorffN,BurgersR,Monsonego-OrnanE,RosenV,andMiosgeN.Adiscoidindomainreceptor1knock-outmouseasanovelmodelforosteoarthritisofthetemporomandibularjoint.CellMolLifeSci 2013;Aug4PMID:23912900PazinDE,GamerLW,CapeloLP,CoxKAandRosenV.37.Genesignatureoftheembryonicmeniscus.JOrthopRes2013;Sept21PMID:24108661WuX,ChimSM,KuekV,LimBS,ChowST,ZhaoJ,Yang38.S,RosenV,TicknerJandXuJ.HtrA1isupregulatedduringRANKL-inducedosteoclastogenesis,andnegativelyregulatesosteoblastdifferentiationandBMP2-inducedSmad1/5/8,ERKandp38phosphorylation.FEBSLetters ,inpress.

20

M E M O

21

miRNAisamemberof〜 22ntnon-cordingRNAandplayacriticalroleinthetissueororgandevelopmentandisalsoassociatedwithhumandiseases.WerecentlyfoundthatmiR-146ishighlyexpressedinrheumatoidarthritissynoviumtoregulateinflammationandthatmiR-140ishighincartilageandimportantfortissuehomeostasisagainstcartilagedegradation.Furthermore,nowwearepreparingseriesofgenomewideanalysistoexaminethemolecularmechanismsofinflammatorygeneexpressionbynon-codingRNAatthelevelofmRNAstabilityortranslation.Here,weintroduceourrecentresultsbynextgenerationsequencingtechniques,whichrevealedseriesofmiRNAsinvolvedinarthritispathogenesis.TogeneratemiRNAknockoutmiceinhigh-throughputway,weappliedTALENsystemtodeletemiRNAgeneinmicegenome,whichsuccessfullyleadsustoanalyzemiRNAfunctionsinvivo.Wealsoshowourstrategy;cellbasedcomprehensivegenescreeningtoscreennewmoleculesthatregulatesinflammatorygeneexpression.CombinationofabovemethodsmayprovidethenovelaspectofRNAregulatorysystemandshouldpromoteourunderstandingofinflammatorydiseasespathogenesis.

Combinatorial approach to identify generegulation of inflammatory signalsat RNA level

Hiroshi Asahara, M.D., Ph.D.

ProfessorDepartmentofSystemsBioMedicine,GraduateSchoolofMedicalandDentalSciences,TokyoMedicalandDentalUniversity

22

Hiroshi Asahara

CURRICULUM VITAE

Education and Position

1992 GraduatefromOkayamaUniversity,Japan1992 Resident,DepartmentofOrthopaedicSurgery, OkayamaSaiseikaiGeneralHospital,Japan1994 GraduateFellow,DepartmentofNeuroscience, InstituteofMolecularandCellularmedicine, OkayamaUniversityMedicalSchool,Japan1995 GraduateFellow,DivisionofRheumatologyand Immunology,InstituteofMedicalScience, St.UniversitySchoolofMedicine,Japan1997 AssistantProfessor,DepartmentofOrthopaedic Surgery,OkayamaUniversityMedicalSchool, Japan1997 PostdoctoralResearchFellow,DepartmentofCell Biology,HarvardMedicalSchool,MA,USA2000 StaffScientist,PeptideBiologyLaboratory,The SalkInstituteforBiologicalStudies,CA,USA2001 Scientist(PrincipalInvestigator),JST,Precursory ResearchforEmbryonicScienceandTechnology, Japan2002 AssistantProfessor,DepartmentofMolecular andExperimentalMedicine,TheScrippsResearch Institute,CA,USA2004 Head,DepartmentofRegenerativeBiologyand Medicine,NationalResearchInstituteforChild HealthandDevelopment,Japan2010 DepartmentHead,DepartmentofSystems BioMedicine,NationalResearchInstituteforChild HealthandDevelopment,Japan2011-pres. VisitingHead,DepartmentofSystemsBioMedicine, NationalResearchInstituteforChildHealthand Development,Japan2011-pres. Professor,DepartmentofSystemsBioMedicine, GraduateSchoolandFacultyofMedicine,Tokyo MedicalandDentalUniversity,Japan

Publications

NakasaT,MiyakiS,OkuboA,HashimotoM,Nishida1.K,OchiM,AsaharaH.ExpressionofmicroRNA-146inrheumatoidarthritissynovialtissue.ArthritisRheum.58(5):1284-1292.(2008)NagataY,NakasaT,MochizukiY,IshikawaM,MiyakiS,2.ShibuyaH,YamasakiK,AdachiN,AsaharaH,OchiM.Inductionofapoptosisinthesynoviumofarthriticmicebyintra-articularinjectionofdouble-strandedmicroRNA-15a.ArthritisRheum.60(9):2677-2683.(2009)MiyakiS,NakasaT,OtsukiS,GroganSP,Higashiyama3.R,InoueA,KatoY,SatoT,LotzMK,AsaharaH.MicroRNA-140isexpressedindifferentiatedhumanarticularchondrocytesandmodulatesIL-1responses.ArthritisRheum.60(9):2723-2730.(2009)ItoT,YadavN,LeeJ,FurumatsuT,YamashitaS,Yoshida4.K,TaniguchiN,HashimotoM,TsuchiyaM,OzakiT,LotzM,BedfordMT,AsaharaH.ArgininemethyltransferaseCARM1/PRMT4regulatesendochondralossification.BMCDevBiol .9:47.(2009)AbeY,OkaA,MizuguchiM,IgarashiT,IshikawaS,5.AburataniH,YokoyamaS,AsaharaH,NagaoK,YamadaM,MiyashitaT.EYA4,deletedinacasewithmiddleinterhemisphericvariantofholoprosencephaly,interactswithSIX3bothphysicallyandfunctionally.HumMutat .30(10):E946-955.(2009)KatoY,MiyakiS,YokoyamaS,OmoriS,InoueA,Horiuchi6.M,AsaharaH.Real-timefunctionalimagingformonitoringmiR-133duringmyogenicdifferentiation.IntJBiochemCellBiol .41(11):2225-2231.(2009)KumagishiK,NishidaK,YamaaiT,MomotaR,MiyakiS,7.HirohataS,NaitoI,AsaharaH,NinomiyaY,OhtsukaA.Adisintegrinandmetalloproteinasewiththrombospondinmotifs9(ADAMTS9)expressionbychondrocytesduringendochondralossification.ArchHistolCytol .72(3):175-185.(2009)YokoyamaS,ItoY,Ueno-KudohH,ShimizuH,Uchibe8.K,AlbiniS,MitsuokaK,MiyakiS,KisoM,NagaiA,HikataT,OsadaT,FukudaN,YamashitaS,HaradaD,MezzanoV,KasaiM,PuriPL,HayashizakiY,OkadoH,HashimotoM,AsaharaH.AsystemsapproachrevealsthatthemyogenesisgenomenetworkisregulatedbythetranscriptionalrepressorRP58.DevCell .17(6):836-848.(2009)ItoY,ToriuchiN,YoshitakaT,Ueno-KudohH,SatoT,9.YokoyamaS,NishidaK,AkimotoT,TakahashiM,MiyakiS,AsaharaH.TheMohawkhomeoboxgeneisacriticalregulatoroftendondifferentiation.ProcNatlAcadSciUSA.107(23):10538-10542.(2010)OtsukiS,GroganSP,MiyakiS,KinoshitaM,AsaharaH,10.LotzMK.TissueneogenesisandSTRO-1expressioninimmatureandmaturearticularcartilage.JOrthopRes .28(1):96-102.(2010)HigashiyamaR,MiyakiS,YamashitaS,YoshitakaT,11.

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Hiroshi Asahara

LindmanG,ItoY,SashoT,TakahashiK,LotzM,AsaharaH.CorrelationbetweenMMP-13andHDAC7expressioninhumankneeosteoarthritis.ModRheumatol .20(1):11-17.(2010)MizutaniR,NakamuraK,YokoyamaS,SanbeA,Kusakawa12.S,MiyamotoY,ToriiT,AsaharaH,OkadoH,YamauchiJ,TanoueA.Developmentalexpressionofsortingnexin3inthemousecentralnervoussystem.GeneExprPatterns .11(1-2):33-40.(2010)MiyakiS,SatoT,InoueA,OtsukiS,ItoY,YokoyamaS,13.KatoY,TakemotoF,NakasaT,YamashitaS,TakadaS,LotzMK,Ueno-KudoH,AsaharaH.MicroRNA-140playsdualrolesbothincartilagedevelopmentandhomeostasis.GenesDev .24(11):1173-1185.(2010)OtsukiS,HansonSR,MiyakiS,GroganSP,KinoshitaM,14.AsaharaH,WongCH,LotzMK.ExtracellularsulfatasessupportcartilagehomeostasisbyregulationBMPandFGFsignalingpathways.ProcNatlAcadSciUSA.107(22):10202-10207.(2010)AsadaM,RauchA,ShimizuH,MaruyamaH,MiyakiS,15.ShibamoriM,KawasomeH,IshiyamaH,TuckermannJ,AsaharaH.DNAbinding-dependentglucocorticoidreceptoractivitypromotesadipogenesisviaKrüppel-likefactor15geneexpression.LabInvest .91(2):203-15.(2011)AwazawaM,UekiK,InabeK,YamauchiT,KubotaN,16.KanekoK,KobayashiM,IwaneA,SasakoT,OkazakiY,OhsugiM,TakamotoI,YamashitaS,AsaharaH,AkiraS,KasugaM,KadowakiT.AdiponectinEnhancesInsulinSensitivitybyIncreasingHepaticIRS-2ExpressionviaaMacrophage-DerivedIL-6-DependentPathway.CellMetab .13(4):401-12.(2011)WadaS,KatoY,OkutsuM,MiyakiS,SuzukiK,Yan17.Z,SchiaffinoS,AsaharaH,UshidaT,AkimotoT.TranslationalsuppressionofatrophicregulatorsbymicroRNA-23aintegratesresistancetoskeletalmuscleatrophy.JBiolChem.286(44):38456-65.(2011)MiyakiS,AsaharaH.MacroviewofmicroRNAfunction18.inosteoarthritis.NatureRevRheumatol .8(9):543-52.(2012)(Review)FukamiM,TsuchiyaT,TakadaS,KanbaraA,AsaharaH,19.IgarashiA,KamiyamaY,NishimuraG,OgataT.ComplexgenomicrearrangementintheSOX95'regioninapatientwithPierreRobinsequenceandhypoplasticleftscapula.AmJMedGenetA.158A(7):1529-34.(2012)YamashitaS,MiyakiS,KatoY,YokoyamaS,SatoT,20.BarrionuevoF,AkiyamaH,SchererG,TakadaS,AsaharaH.L-Sox5andSox6proteinsenhancechondrogenicmiR-140microRNAexpressionbystrengtheningdimericSox9activity.JBiolChem287(26),22206-22215(2012)UchibeK,ShimizuH,YokoyamaS,KubokiT,Asahara21.H.Identificationofnoveltranscription-regulatinggenesexpressedduringmurinemolardevelopment.DevDyn .241(7):1217-26.(2012)FurumatsuT,AsaharaH.Chromatinassemblyandinvitro22.

transcriptionanalysesforevaluationofindividualproteinactivitiesinmulticomponenttranscriptionalcomplexes.MethodsMolBiol .977:193-202.(2013)(Review)GibsonG,AsaharaH.microRNAsandcartilage.23. JOrthopRes .Sep;31(9):1333-44.(2013)(Review)WatanabeT,OyamaT,AsadaM,HaradaD,ItoY,24.InagawaM,SuzukiY,SuganoS,KatsubeK,KarsentyG,KomoriT,KitagawaM,AsaharaH.MAML1enhancesthetranscriptionalactivityofRunx2andplaysaroleinbonedevelopment.PLOSGenetics .9(1):e1003132.(2013)MatsukawaT,SakaiT,YonezawaT,HiraiwaH,Hamada25.T,NakashimaM,OnoY,IshizukaS,NakaharaH,LotzMK,AsaharaH,IshiguroN.MicroRNA-125bregulatestheexpressionofaggrecanase-1(ADAMTS-4)inhumanosteoarthriticchondrocytes.ArthritisResTher .15(1):R28.(2013)HasegawaA,NakaharaH,KinoshitaM,AsaharaH,Koziol26.J,LotzMK.Cellularandextracellularmatrixchangesinanteriorcruciateligamentsduringhumankneeagingandosteoarthritis.ArthritisResTher .15(1):R29(2013)RakoczyJ,Fernandez-ValverdeSL,GlazovEA,Wainwright27.EN,SatoT,TakadaS,CombesAN,KorbieDJ,MillerD,GrimmondSM,LittleMH,AsaharaH,MattickJS,TaftRJ,WilhelmD.MicroRNAs-140-5p/140-3pmodulateLeydigcellnumbersinthedevelopingmousetestis.BiolReprod .88(6):143.(2013)NakaharaH,HasegawaA,OtabeK,AyabeF,Matsukawa28.T,OnizukaN,ItoY,OzakiT,LotzMK,AsaharaH.Transcriptionfactormohawkandthepathogenesisofhumananteriorcruciateligamentdegradation.ArthritisRheum.65(8):2081-9.(2013)YoshitakaT,KawaiA,MiyakiS,NumotoK,KikutaK,29.OzakiT,LotzM,AsaharaH.AnalysisofmicroRNAsexpressionsinchondrosarcoma.JOrthopRes .inpressShimizuH,KuboA,UchibeK,HashimotoM,YokoyamaS,30.TakadaS,MitsuokaK,AsaharaH,TheAEROSystem:a3D-likeApproachforRecordingGeneExpressionPatternsintheWholeMouseEmbryo,PLOSOne .inpressTakadaS,SatoT,ItoY,YamashitaS,KatoT,Kawasumi31.M,Kanai-AzumaM,IgarashiA,KatoT,TamanoM,AsaharaH.TargetedgenedeletionofmiRNAsinmicebyTALENsystem,PLOSOne .inpressOnizukaN,ItoY,InagawaM,NakaharaH,TakadaS,32.LotzMK,ToyamaY,AsaharaH.TheMohawkhomeoboxtranscriptionfactorregulatesthedifferentiationoftendonsandvolarplates.JOrthopScience .inpressKatoT,MiyataK,SonobeM,YamashitaS,TamanoM,33.MiuraK,KanaiY,MiyamotoS,SakumaT,YamamotoT,InuiM,KikusuiT,AsaharaH*,andTakadaS*.ProductionofSryknockoutmouseusingTALENviaoocyteinjection.ScientificRep .inpress(*equallycontributedcorrespondingauthors)

24

M E M O

25

Infantilehemangiomasarethemostcommontumorsofinfancy.Typically,theyappearwithinafewdays/weeksafterbirthandgrowrapidlyduringafewmonths(proliferativephase).Afteraboutayeartheystarttoinvolute(involutionphase)andafteraperiodofseveralmonths/years,theyarereplacedbyfibro-fattytissue.Hemangiomasaremorecommoninfemalesthaninmalesandtheyoccurmostfrequentlyintheheadandneckregionofaffectedinfants.Ourstudieshavedemonstratedthattherapidproliferationofinfantilehemangiomasiscausedbydefectsintheregulationofvascularendothelialgrowthfactorreceptor1(VEGFR1)inendothelialcellsofthetumor.WhennormalendothelialcellsarestimulatedbyVEGForactivatedbybindingtoextracellularmatrix,expressionoftheVEGFdecoyreceptorVEGFR1isstimulatedandVEGFR2-dependentsignalingconstrained.Incontrast,hemangiomaendothelialcellsintheirproliferativephaseexhibitconstitutivelowlevelsofVEGFR1expressionandhighlevelsofVEGF-dependentVEGFR2signaltransduction.Thisisaconsequenceofdefectsthatresultinlocallossoffunctionofacellsurfaceintegrin-likereceptorknownasAnthraxtoxinreceptor1(ANTXR1)/Tumorendothelialmarker8(TEM8)inhemangiomaendothelialcells.MicethatarehomozygousforTem8nullalleleshavechangesindermalbloodvesselsthataresimilartothoseseeninhemangiomatumors.Inaddition,themiceexhibitgrowthretardation,bonelossandcraniofacialdefectssimilartothoseseeninpatientswiththerecessiveGAPOsyndrome.Patientswiththissyndromearehomozygousforloss-of-functionmutationsinTEM8andexhibitgrowthretardation,alopecia,pseudo-anodontiaandopticatrophy,aswellasothercraniofacialdefects,bonelossandhemangioma.TEM8isthereforeacriticalregulatorofvascularendothelialandosteoblasticfunctionsinadditiontocontrollingtheactivitiesofgrowthplatesandsynchondrosesattheskullbase.

References:1. Jinnin,M.,etal.,SuppressedNFAT-dependentVEGFR1expressionandconstitutiveVEGFR2signalingininfantile

hemangioma .Nat.Med.,2008.14(11):p.1236-1246.

Craniofacial dysmorphogenesis andinfantile hemangioma – the antrax toxin receptor connection

Bjorn R. Olsen, M.D., Ph.D.

ProfessorDepartmentofOralandDevelopmentalBiology,HarvardSchoolofDentalMedicine

26

Bjorn R. Olsen

CURRICULUM VITAE

Education

1967 Ph.D.UniversityofOslo,Norway1967 M.D.UniversityofOsloMedicalSchool,Norway

Position

1990-1993 Chairman,PrograminCellandDevelopmental Biology,HarvardMedicalSchool,Boston,MA1993- HerseyProfessorofCellBiology,Departmentof CellBiology,HarvardMedicalSchool,Boston,MA1996-2002 ProfessorofOralBiology,HarvardSchoolof DentalMedicine,Boston,MA1996-2002 Chairman,Harvard-ForsythDepartmentofOral Biology,HarvardSchoolofDentalMedicine, Boston,MA2002- ProfessorofOralandDevelopmentalBiology, HarvardSchoolofDentalMedicine,Boston,MA2002-2005 Chairman,DepartmentofOralandDevelopmental Biology,HarvardSchoolofDentalMedicine, Boston,MA2005- DeanforResearch,HarvardSchoolofDental Medicine,Boston,MA

Award and Honors

2000 HonoraryDoctorofScienceDegree,Universityof MedicineandDentistryofNewJersey2000 HonoraryDoctorofScienceDegree,UniversityofOslo, Norway2001 DistinguishedFacultyAward,HarvardSchoolofDental Medicine2006 H.C.JacobæusPrizeandlecturer,H.C.Jacobæus’ ForelæsningerFoundation,Sweden2006 Member,ScanBaltAcademy2006 SeniorResearchPrize,AmericanSocietyofMatrix Biology2007 Co-chairman,GordonResearchConference“Cartilage Biology&Pathology”2009 Chairman,GordonResearchConference“Cartilage Biology&Pathology”2009 Co-chairman,GordonResearchConference“Bonesand Teeth”2010 IADRDistinguishedScientistAwardforCraniofacial BiologyResearch2010 ISMBDistinguishedInvestigatorAward2010 AmericanAssociationfortheAdvancementofScience Fellow2011 HenryGrayAward,AmericanAssociationof Anatomists2011 Chairman,GordonResearchConference“Bonesand Teeth”2011 Fellow,AmericanAssociationofAnatomists2011 HonoraryDoctorDegree,OkayamaUniversity,Japan

Publications

Sun,X-D,Jeng,L,Bolliet,C,Olsen,BR,Spector,M.Non-1.viralendostatinplasmidtransfectionofmesenchymalstemcellsviacollagenscaffolds.Biomaterials ,200930:1222-1231.Hou,B,Kolpakova-Hart,E,Fukai,N,Wu,K,Olsen,BR.2.Thepolycystickidneydisease1(Pkd1)geneisrequiredfortheresponsesofosteochondroprogenitorcellstomidpalatalsutureexpansioninmice.Bone ,2009,44:1121-1133.PMC2680722Faye,C,Chautard,E,Olsen,BR,Ricard-Blum,S.Thefirst3.draftoftheendostatininteractionnetwork.J.Biol.Chem.2009,284:22041-22047.PMC2755928Galli,C,Fu,Q,Wang,WF,Olsen,BR,Manolagas,SC,Jilka,R,4.andO’Brien,CA.CommitmenttotheosteoblastlineageisnotrequiredforRANKLgeneexpression.JBiolChem2009,284:12654-12662.PMC2675994Olsen,BR.Geneticsofhumanskeletaldisease.In:5. TheSkeletalSystem,ed.OlivierPourquié.ColdSpringHarborLaboratoryPress.2009,pp.317-340.Boye,E,Jinnin,M,Olsen,BR.Infantilehemangioma–6.challenges,newinsightsandtherapeuticpromise.JCraniofSurg ,2009,20:1-7.PMID:19190505.Boye,E,Olsen,BR.Signalingmechanismsininfantile7.hemangioma.CurrOpinHematol .2009,16:202–208.PMC2895461Xu,L,PolurI,LimC,ServaisJM,DobeckJ,LiY,Olsen8.BR.Early-onsetosteoarthritisofmousetemporomandibularjointinducedbypartialdiscectomy.OsteoarthrCartilage2009,17:917-922.PMC2941347Gebauer,JM,Keene,DR,Olsen,BR,Sorokin,LM,Paulsson,9.M,Wagener,R.MouseAMACO,akidneyandskinbasementmembraneassociatedmoleculethatmediatesRGD-dependentcellattachment.MatrixBiol 2009,28:456-462.(NoBROgrant)OlsenBR.Whenprimaryciliafail.10. MatrixBiol 2009;28:443-4.Review.PMID:19939339.Mukherjee,PM,Wang,CJ,Chen,I-P,Jafarov,T,Olsen,11.BR,Yueki,Y,Reichenberger,EJ.CherubismgeneSh3bp2isimportantforoptimalboneformation,osteoblastdifferentiationandfunction.AmJOrthodDentofacialOrthop.2010,138:140.e1-140.e11.PMC3268358Besschetnova,TY,Kolpakova-Hart,E,Guan,Y,Zhou,J,12.Olsen,BR,Shah,JV.Identificationofsignalingpathwaysregulatingprimaryciliumlengthandflow-mediatedadaptation.Curr.Biol.,2010,20:182-187.PMC2990526.Harnagea,C,Vallie`res,M,Pfeffer,CP,Wu,D,Olsen,BR,13.Pignolet,A,Legare,F,Gruverman,A.Two-dimensionalnanoscalestructuralandfunctionalimaginginindividualcollagentype-Ifibrils.BiophysJ ,2010,98:3070-3077.PMC2884257Nicolae,C,Olsen,BR.Unexpectedmatrixdiseasesand14.noveltherapeuticstrategies.CellTissueRes.2010,339:155–165.ReviewJinnin,M,Ishihara,T,Boye,E,Olsen,BR.Recentprogress15.

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Bjorn R. Olsen

instudiesofinfantilehemangioma.JDerm.2010,37:283-298.Review.Faye,C,Inforzato,A,Bignon,M,Hartmann,DJ,16.Muller,L,Ballut,LOlsen,BR,Day,AJ,Ricard-Blum,S.Transglutaminase-2:anewendostatinpartnerintheextracellularmatrixofendothelialcells.Biochem.J .2010,427:467-475.PMCID:PMC2876729.Jeng,L,Olsen,BR,Spector,M.Engineeringendostatin-17.producingcartilaginousconstructsforcartilagerepairusingnon-viraltransfectionofchondrocyte-andmesenchymalstemcell(msc)-seededcollagenscaffolds.TissEngPartA,2010,16:3011-3021.PMC3189205Clarkin,C,Olsen,BR.Onbone-formingcellsandblood18.vesselsinbonedevelopment.Preview.CellMetabol ,2010,12:314-316.Preview.PMC3031181Medici,D,Shore,EM,Lounev,VY,Kaplan,FS,Kalluri,R,19.Olsen,BR.Conversionofvascularendothelialcellsintomultipotentstem-likecells.NatMed2010,16:1400-1406.PMC3209716Mauney,J,Olsen,BR,VollochV.Matrixremodelingas20.stemcellrecruitmentevent:anovelinvitromodelforhomingofhumanbonemarrowstromalcellstothesiteofinjuryshowscrucialroleofextracellularcollagenmatrix.MatrixBiol .2010,29:657-663.Medici,D,Olsen,BR.Transformingbloodvesselsintobone.21.CellCycle2011,10:362-363.Freefulltexthttp://www.landesbioscience.com/journals/cc/article/14519/PfefferCP,OlsenBR,GanikhanovF,LegareF.Imaging22.skeletalmuscleusingharmonicgenerationandcoherentanti-StokesRamanscatteringmicroscopy.BiomedOptExpress2011;27:1366-76.PMC3087593BerendsenA,OlsenBR.Tankyraselosesitsgripon23.SH3BP2incherubism.Cell 2011;147:1222-3.PMID:22153068.Jeng,L,Olsen,BR,Spector,M.Engineeringendostatin-24.expressingcartilaginousconstructsusinginjectablebiopolymerhydrogels.ActaBiomat ,2012,8:2203-2212.PMID:22370447(noNIHgrant).Ionescu,I,Kozhemyakina,E,Nicolae,C,Kaestner,KH,25.Olsen,BR,Lassar,AB.FoxAfamilymembersarecrucialregulatorsofthehypertrophicchondrocytedifferentiationprogram.DevCell 2012;22:927-39.PMC3356573Papadaki,ME,Lietman,SA,Levine,MA,Olsen,BR,Kaban26.LB,Reichenberger,EJ.Cherubism:BestclinicalPractice..OrphanetJRareDis .2012,7Suppl1:S6.PMC3359956Reichenberger,EJ,Levine,MA,Olsen,BR,Papadaki,ME,27.Lietman,SA.TheroleofSH3BP2inthepathophysiologyofcherubism.OrphanetJRareDis .2012,7Suppl1:S5.PMC3359958Jumlongras,D,Lachke,SA,O'Connell,DJ,Aboukhalil,28.A,Li,X,Choe,SE,Ho,JW,Turbe-Doan,A,Robertson,EA,Olsen,BR,Bulyk,ML,Amendt,BA,andMaas,RL.AnEvolutionarilyConservedEnhancerRegulatesBmp4ExpressioninDevelopingIncisorandLimbBud.PLoS

ONE2012;7:e38568.PMC3373496Medici,D,Olsen,BR.Theroleofendothelial-mesenchymal29.transitioninheterotopicossification.JBMR2012;27:1619-22.PMID:22806925.Review.KatebiN,Kolpakova-HartE,LinCY,OlsenBR.Themouse30.palateanditscellularresponsestomidpalatalsutureexpansionforces.OrthoCraniofacRes2012;15:148-158.NIHMS512089Medici,D.andOlsen,B.R.(2012).Transformationof31.VascularEndothelialCellsintoMultipotentStem-LikeCells:RoleoftheActivin-LikeKinase-2Receptor.StemCellsandCancerStemCells:TherapeuticApplicationsinDiseaseandInjury.Springer.Volume8,Chapter19Medici,D,Olsen,BR.Rapamycininhibitsproliferationof32.hemangiomaendothelialcellsbyreducingHIF-1-dependentexpressionofVEGF.PLoSONE,2012;7(8):e42913.PMC3416768Liu,Y,Berendsen,A,Jia,S,Lotinun,S,Baron,R,Ferrara,33.N,Olsen,BR.IntracellularVEGFregulatesthebalancebetweenosteoblastandadipocytedifferentiation.JCI ,2012;122:3101-3112.PMC3428080Adams,A,McBratney-Owen,B,Newby,B,Bowen,M,34.Olsen,BR,Warman,ML.PresphenoidalsynchondrosisfusioninDBA/2Jmice.MammalGenome,2013,24:54-62.PMC3560942Olsen,BR.Matrixmoleculesandtheirligands.In:35.PrinciplesofTissueEngineering.R.G.(Lanza,R,Langer,R,andVacanti,J.ed).4nded.,inpress,AcademicPress,pp.Duncan,MD,Yang,C,Tanjore,H,Boyle,PM,Keskin,D,36.Sugimoto,H,Zeisberg,M,Olsen,BR,Kalluri,R.TypeXVIIIcollagenisessentialforsurvivalduringacuteliverinjury.DisModelMech ,2013Apr4.Epubaheadofprint.Freefulltext.Toh,WS,Foldager,CB,Olsen,BR,Spector,M.Basement37.membranemoleculeexpressionattendanttochondrogenesisbynucleuspulposuscellsandmesenchymalstemcells.JOrthopRes ,2013,Epubaheadofprint.(noNIHgrant)Volloch,V,Olsen,BR.Whycellularstresssuppresses38.adipogenesisinskeletaltissue,butisineffectiveinadiposetissue:Controlofmesenchymalcelldifferentiationviaintegrinbindingsitesinextracellularmatrices.MatBiol,2013Jun18.doi:pii:S0945-053X(13)00091-7.10.1016/j.matbio.2013.06.001.[Epubaheadofprint].NIHMSID#504634Berendsen,AD,Olsen,BR.Osteoblast-adipocytelineage39.plasticityintissuedevelopment,maintenanceandpathology.Review.CellMolLifeSci.inpress.NIHMS514666Olsen,BR,Ehrlich,YH,Yalak.Extracellularprotein40.phosphorylationrevisited.Submittedforpublication.Opinion.Liu,Y,Olsen,BR.DistinctVEGFfunctionsduringbone41.developmentandhomeostasis.Submittedforpublication.Berendsen,AD,Olsen,BR.HowVascularEndothelial42.GrowthFactorA(VEGF)RegulatesDifferentiationofMesenchymalStemCells.JHC,Submittedforpublication.Review.NIHBonegrantsupport.

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M E M O

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Accordingtoourpreviousstudies,thenumberofmesenchymalstemcells(MSCs)insynovialfluidincreaseinkneeswithanteriorcruciateligamentinjury(Rheumatology2008),meniscusinjury(ClinOrthopRelatRes2014),andosteoarthritis(JOrthopRes2011).ThemorphologyandgeneprofilesinsynovialfluidMSCsweremoresimilartothoseinsynovialMSCsthaninbonemarrowMSCs.Principalcomponentanalysisofgeneprofilesforvariousmesenchymaltissues-derivedMSCsandchondrocytesdemonstratedthatMSCsfromintraarticulartissuesandchondrocyteswereclosertoeachotherthanMSCsfromextraarticulartissues(JOrthopRes2008).SynoviummaybeareservoirforMSCstocontributetotheintraarticulartissuerepair.Afterintraarticulartissues,suchascartilage,meniscus,andligament,areinjured,MSCsmaybemobilizedfromsynoviumtosynovialfluid,adheretotheinjuredsite,andcontributetotherepair.However,thenumberofMSCsislimited,therefore,theinjuredtissuescannotbehealedinthenaturalcourse.TransplantationofenoughnumberofsynovialMSCstotheinjuredtissuesmaypromoteintraarticulartissuehealing.

MSCsareattractivecellsourceforcartilageandmeniscusregeneration.OurinvitroandinvivochondrogenicassaydemonstratedthatsynovialandbonemarrowMSCshadahigherchondrogenicabilitythanadiposeandmuscleMSCs(ArthritisRheum2005,CellTissueRes2007,CellTissueRes2008).HumansynovialMSCsexpandedmoreinhumanserumthanbonemarrowMSCs(ArthritisRheum2008).Inrat,rabbit,andpigstudies,transplantationofsynovialMSCspromotedcartilageandmeniscusregeneration(StemCells2007,StemCells2009,Cytotherapy2012,OsteoarthritisCartilage2012,JBoneJointSurgAm2012,BiochemBiophysResCommun2013).

Currentcelltherapyforcartilageandmeniscusregenerationrequiresinvasiveprocedures.WehavedevelopedanovelimplantationprocedurewithsynovialMSCs.CartilageormeniscusdefectisfilledwithsynovialMSCsuspensionfor10minutes.Accordingtoourinvitroandinvivostudies,morethan60%cellsadheredtothedefect,andpromotedcartilageandmeniscusregeneration(ArthritisResTher2008,JOrthopRes2013).

Wearecurrentlydoingclinicaltrialforcartilageregeneration.AllpatientshavetheircartilagedefectsfilledwithsynovialMSCsarthroscopically.FavorableresultsareobtainedbyMRIimaginginmanycases,bysecondlookarthroscopies,andbybiopsies.Ourmethodhassuchadvantagesthatnoperiostealcoverageorscaffoldwererequiredandthattransplantationispossiblearthroscopically.Wearealsotryingtoregenerateosteoarthritisofthekneewithosteotomyormeniscuscentralization(ArthroscTech2012)byusingsynovialMSCs.WearegoingtostartanotherclinicaltrialformeniscustreatmentwithsynovialMSCs.

Cartilage and meniscus regeneration withsynovial stem cells

Ichiro Sekiya, M.D., Ph.D.

ProfessorCenterforStemCellandRegenerativeMedicine,TokyoMedicalandDentalUniversity

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Ichiro Sekiya

CURRICULUM VITAE

Education

1984-1990 MD,SchoolofMedicine,TokyoMedicalandDental University1996-2000 PhD,SchoolofMedicine,TokyoMedicaland DentalUniversity

Position

1990-1996 OrthopedicSurgeryat10hospitalsinJapan2000-2000 CenterforGeneTherapy,MCPHahnemann University,Philadelphia,USA2000-2002 CenterforGeneTherapy,TulaneUniversity,New Orleans,USA2002-2008 TokyoMedicalandDentalUniversity2008-2011 AssociateProfessor,CartilageRegeneration TokyoMedicalandDentalUniversity2011-2013 Professor,CartilageRegeneration TokyoMedicalandDentalUniversity2013- Director/Professor CenterforStemCellandRegenerativeMedicine TokyoMedicalandDentalUniversity2013- Professor AppliedRegenerativeMedicine TokyoMedicalandDentalUniversity

Publications

Transplantationofneuronalcellsinducedfromhuman1.mesenchymalstemcellsimprovesneurologicalfunctionsafterstrokewithoutcellfusion.XuH,MikiK,IshibashiS,InoueJ,SunL,EndoS,SekiyaI,MunetaT,InazawaJ,DezawaM,MizusawaH.JNeurosciRes.2010;88(16):3598-609.RepairofolecranonfracturesusingfiberWirewithout2.metallicimplants:reportoftwocases.NimuraA,NakagawaT,WakabayashiY,SekiyaI,OkawaA,MunetaT.JOrthopSurgRes.2010Oct12;5:73.EffectofCTslicethicknessonaccuracyofimplant3.positioninginnavigatedtotalhiparthroplasty.HirasawaN,MatsubaraM,IshiiK,HagioS,OkudaN,SekiyaI,MunetaT.ComputAidedSurg.2010Nov2.Intradiscaltransplantationofsynovialmesenchymalstem4.cellspreventsintervertebraldiscdegenerationthroughsuppressionofmatrixmetalloproteinase-relatedgenesinnucleuspulposuscellsinrabbits.MiyamotoT,MunetaT,TabuchiT,MatsumotoK,SaitoH,TsujiK,SekiyaI.ArthritisResTher.2010;12(6):R206.Hipacetabulardysplasiaandjointlaxityoffemaleanterior5.cruciateligament-injuredpatients.YamazakiJ,MunetaT,JuYJ,MoritoT,OkuwakiT,SekiyaI.AmJSportsMed.2011Feb;39(2):410-4.Radiographicdescriptionoffemoraltunnelplacement6.expressedasintercondylarclocktimeindouble-bundleanteriorcruciateligamentreconstruction.YamazakiJ,MunetaT,KogaH,SekiyaI,JuYJ,MoritoT,YagishitaK.KneeSurgSportsTraumatolArthrosc.2011Mar;19(3):418-23.Ketoprofenabsorptionbymuscleandtendonaftertopical7.ororaladministrationinpatientsundergoinganteriorcruciateligamentreconstruction.SekiyaI,MoritoT,HaraK,YamazakiJ,JuYJ,YagishitaK,MochizukiT,TsujiK,MunetaT.AAPSPharmSciTech.2010Mar;11(1):154-8.BMP-7inhibitscartilagedegenerationthroughsuppression8.ofinflammationinratzymosan-inducedarthritis.TakahashiT,MunetaT,TsujiK,SekiyaI.CellTissueRes.2011May;344(2):321-32.High-flexPosteriorCruciate-RetainingvsPosterior9.Cruciate-SubstitutingDesignsinSimultaneousBilateralTotalKneeArthroplastyAProspective,RandomizedStudy.YagishitaK,MunetaT,JuYJ,MoritoT,YamazakiJ,SekiyaI.JArthroplasty.2012Mar;27(3):368-74.Effectsofdifferentinitialbundletensioningstrategieson10.theoutcomeofdouble-bundleACLreconstruction:acohortstudy.MunetaT,KogaH,JuYJ,YagishitaK,SekiyaI.SportsMedArthroscRehabilTherTechnol.2011Jul28;3:15.Surgicalmanagementofgrade3medialkneeinjuries11.combinedwithcruciateligamentinjuries.KogaH,MunetaT,YagishitaK,JuYJ,SekiyaI.KneeSurgSportsTraumatolArthrosc.2012Jan;20(1):88-94.Humanmesenchymalstemcellsinsynovialfluidincrease12.inthekneewithdegeneratedcartilageandosteoarthritis.

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SekiyaI,OjimaM,SuzukiS,YamagaM,HorieM,KogaH,TsujiK,MiyaguchiK,OgishimaS,TanakaH,MunetaT.JOrthopRes.2012Jun;30(6):943-9.TheEffectofGraftFixationAnglesonAnteroposterior13.andRotationalKneeLaxityinDouble-BundleAnteriorCruciateLigamentReconstruction:EvaluationUsingComputerizedNavigation.KogaH,MunetaT,YagishitaK,JuYJ,SekiyaI.AmJSportsMed.2012Mar;40(3):615-23.Epub2011Nov22.Arthroscopic,histologicalandMRIanalysesofcartilage14.repairafteraminimallyinvasivemethodoftransplantationofallogeneicsynovialmesenchymalstromalcellsintocartilagedefectsinpigs.NakamuraT,SekiyaI,MunetaT,HatsushikaD,HorieM,TsujiK,KawarasakiT,WatanabeA,HishikawaS,FujimotoY,TanakaH,KobayashiE.Cytotherapy.2012Mar;14(3):327-38.Implantationofallogenicsynovialstemcellspromotes15.meniscalregenerationinarabbitmeniscaldefectmodel.HorieM,DriscollMD,SampsonHW,SekiyaI,CaroomCT,ProckopDJ,ThomasDB.JBoneJointSurgAm.2012Apr18;94(8):701-12.Hyaluronaninjectiontherapyforathleticpatientswith16.patellartendinopathy.MunetaT,KogaH,JuYJ,MochizukiT,SekiyaI.JOrthopSci.2012Apr17.Remnantvolumeofanteriorcruciateligamentcorrelates17.preoperativepatients'statusandpostoperativeoutcome.MunetaT,KogaH,JuYJ,HorieM,NakamuraT,SekiyaI.KneeSurgSportsTraumatolArthrosc.2012Apr28.Intra-articularinjectionofhumanmesenchymalstem18.cells(MSCs)promoteratmeniscalregenerationbybeingactivatedtoexpressIndianhedgehogthatenhancesexpressionoftypeIIcollagen.HorieM,ChoiH,LeeRH,RegerRL,YlostaloJ,MunetaT,SekiyaI,ProckopDJ.OsteoarthritisCartilage.2012Oct;20(10):1197-207.Isolationandcharacterizationofmultipotential19.mesenchymalcellsfromthemousesynoviumFutamiI,IshijimaM,KanekoH,TsujiK,Ichikawa-TomikawaN,SadatsukiR,MunetaT,Arikawa-HirasawaE,SekiyaI,KanekoK.PLoSOne.2012;7(9):e45517.Incidenceandriskfactorsformedialtibialstresssyndrome20.andtibialstressfractureinhighschoolrunners.YagiS,MunetaT,SekiyaI.KneeSurgSportsTraumatolArthrosc.2012Aug9.Osteopontinlevelinsynovialfluidisassociatedwiththe21.severityofjointpainandcartilagedegradationafteranteriorcruciateligamentruptureYamagaM,TsujiK,MiyatakeK,YamadaJ,AbulaK,JuYJ,SekiyaI,MunetaT.PLoSOne.2012;7(11):e49014Propertiesandusefulnessofaggregatesofsynovial22.mesenchymalstemcellsasasourceforcartilageregeneration.SuzukiS,MunetaT,TsujiK,IchinoseS,MakinoH,UmezawaA,SekiyaI.ArthritisResTher.2012Jun7;14(3):R136.Arthroscopiccentralizationofanextrudedlateral23.

meniscus.KogaH,MunetaT,YagishitaK,WatanabeT,MochizukiT,HorieM,NakamuraT,OkawaA,SekiyaI.ArthroscTech.2012Oct22;1(2):e209-12.IntraoperativeJointGapsAffectPostoperativeRangeof24.MotioninTKAsWithPosterior-stabilizedProstheses.WatanabeT,MunetaT,SekiyaI,BanksSA.ClinOrthopRelatRes.2013Apr;471(4):1326-33.HumanYKL39(chitinase3-likeprotein2),anosteoarthritis-25.associatedgene,enhancesproliferationandtypeIIcollagenexpressioninATDC5cells.MiyatakeK,TsujiK,YamagaM,YamadaJ,MatsukuraY,AbulaK,SekiyaI,MunetaT.BiochemBiophysResCommun.2013Feb1;431(1):52-7.Comparativesequentialmorphologicalanalysesduringin26.vitrochondrogenesisandosteogenesisofmesenchymalstemcellsembeddedincollagengels.IchinoseS,TagamiM,MunetaT,MukohyamaH,SekiyaI.MedMolMorphol.2013Mar;46(1):24-33.Thekinematicanalysisoffemalesubjectsafterdouble-27.bundleanteriorcruciateligamentreconstructionduringsingle-legsquatting.YamazakiJ,MunetaT,JuYJ,KogaH,MoritoT,SekiyaI.JOrthopSci.2013Mar;18(2):284-9.EffectofPosterolateralBundleGraftFixationAngles28.onGraftTensionCurvesandLoadSharinginDouble-BundleAnteriorCruciateLigamentReconstructionUsingaTranstibialDrillingTechnique.KogaH,MunetaT,YagishitaK,JuYJ,MochizukiT,HorieM,NakamuraT,OkawaA,SekiyaI.Arthroscopy.2013Mar;29(3):529-38.Isolationandcharacterizationofhumanmesenchymalstem29.cellsderivedfromshouldertissuesinvolvedinrotatorcufftears.UtsunomiyaH,UchidaS,SekiyaI,SakaiA,MorideraK,NakamuraT.AmJSportsMed.2013Mar;41(3):657-68.Synovialstemcellsinmusculoskeletalregeneration.Atesok30.K,DoralMN,BilgeO,SekiyaI.JAmAcadOrthopSurg.2013Apr;21(4):258-9.5.IntraarticularInjectionofSynovialStemCellsPromotes31.MeniscalRegenerationinaRabbitMassiveMeniscalDefectModel.HatsushikaD,MunetaT,HorieM,KogaH,TsujiK,SekiyaI.JOrthopRes.2013Sep;31(9):1354-9.Transplantationofaggregatesofsynovialmesenchymal32.stemcellsregeneratesmeniscusmoreeffectivelyinaratmassivemeniscaldefect.KatagiriH,MunetaT,TsujiK,HorieM,KogaH,OzekiN,KobayashiE,SekiyaI.BiochemBiophysResCommun.2013Jun14;435(4):603-9.Effectoffemoraltunnelpositionongrafttensioncurves33.andkneestabilityinanatomicdouble-bundleanteriorcruciateligamentreconstruction.KogaH,MunetaT,YagishitaK,WatanabeT,MochizukiT,HorieM,NakamuraT,SekiyaI.KneeSurgSportsTraumatolArthrosc.2013Sep24.TransplantationofAchillestendontreatedwithBMP-734.promotesmeniscusregenerationinaratmassivemeniscusdefectmodel.OzekiN,MunetaT,KogaH,KatagiriH,OtabeK,OkunoM,TsujiK,KobayashiE,MatsumotoK,SaitoH,SaitoT,SekiyaI.ArthritisRheum.2013Jul29.

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M E M O

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Thepaceoffracturerepairslowswithaging,butthecriticalmediatorsinthisprocessarenowknown.Herewedeterminedtherelativecontributionofmesenchymalprogenitorcellage,hematopoieticcellage,andcirculatingfactorsoninvivoboneregenerationandinvitroosteoblastdifferentiation.Exposuretoyouthfulcirculationbyheterochronicparabiosisreversedtheslowedfracturerepaircharacteristicofaginginmice.Thisrejuvenationresultedinincreasedmineralizationandosteoblasticactivityofoldmesenchymalprogenitorcells.Todetermineifcirculatingcellsplayedaroleinthisrejuvenation,weexaminedtheabilityofoldandyoungbonemarrowcellsinwhichwecoulddeletecellswithosteogenicpotentialtoenhancetheosteogeniccapacityofoldbonemarrowcells.Conditionedmediafromanadherentpopulationofyoungcellsrejuvenatedtheosteogeneiccapacityofoldcells.Similarly,engraftmentofyounghematopoieticcellsintooldanimalsrescuedfracturerepairandosteogenicpotentialinamechanismthatdidnotrequireosteoblastsfromthedonoranimal.β-cateninsignaling,apathwayimportantinboneregenerationandosteoblastdifferentiation,wasshowntobemodulatedduringrejuvenation.Reductionofβ-cateninsignalingduringearlyfracturerepairimprovedboneregenerationinoldmice.Thesedatademonstratethatthecirculatorysystemcarrieswithinita“youthfactor"thatisabletorejuvenatebonerepairandosteoblastdifferentiationthroughmodulationofβ-catenin.Thisdataraisesthepossibilitythatagentsthatmodulateβ-catenincanimprovethequalityofbonerepairintheagingpopulation.

Fracture repair and beta-catenin in agingand disease : implications for an approachto therapy

Benjamin A. Alman, M.D.

ProfessorDepartmentofOrthopaedicSurgery,UniversityofToronto

34

Benjamin A. Alman

CURRICULUM VITAE

Education

HighSchool NortheastHighschool,Philadelphia, PennsylvaniaUSA1978HighSchoolDiplomaCollege UniversityofPennsylvania,Philadelphia, Pennsylvania,USA1982.BSc,Material ScienceandEngineering(SummacumLaude)ProfessionorgraduateSchool JeffersonMedicalCollege,ThomasJefferson University,Philadelphia,Pennsylvania,USA1986 MD

Position

2002-present UniversityofToronto,Toronto,Ontario,Canada, Member,CollaborativeProgramin DevelopmentalBiology2002-present UniversityofToronto,Toronto,Ontario,Canada, Member,CollaborativePrograminMolecular Medicine2002-present TheResearchInstituteoftheHospitalforSick Children,Toronto,Ontario,Canada,Senior Scientist,PrograminDevelopmentalandStem CellBiology2004-present UniversityofToronto,Toronto,Ontario,Canada, ViceChairResearch,Surgery2004-present UniversityofToronto,Toronto,Ontario,Canada, Professor,Surgery2004-present UniversityofToronto,Toronto,Ontario,Canada, Professor,LaboratoryMedicineandPathobiology2005-present TheHospitalforSickChildren,Toronto,Ontario, Canada,Head,DivisionofOrthopaedics2006–present UniversityofToronto,Toronto,Ontario,Canada, A.J.LatnerProfessorandChairofOrthopaedics2011-present UniversityofToronto,Toronto,Ontario,Canada, InterimDirector,TorontoMusculoskeletal Centre(Extra-DepartmentalUnit)

Award and Honors

2013 2013ASEAwardforExcellenceinInnovation, AssociationforSurgicalEducation,LosAngles, California,UnitedStates.(Distinction)Awardedtothe TorontoOrthopaedicBootCamp.Annualawardfor exemplaryperformanceinsurgicaleducationwiththe intenttorecognizenovelideasand/ormethodsfor improvingteachingandlearning.2012 FellowshipintheCanadianAcademyofHealth Sciences,CanadianAcademyofHealthSciences, Ottawa,Ontario,Canada.FellowsoftheAcademy areelectedonthebasisoftheirdemonstrated leadership,creativity,distinctivecompetenciesand commitmenttoadvanceacademichealthsciences.

Publications

ChenY,AlmanBA.Wntpathway,anessentialroleinbone1.regeneration.JournalofCellularBiochemistry.2009:106(3):353-362.GurungA,UddinF,HillRP,FergusonPC,AlmanBA.2.Beta-cateninisamediatoroftheresponseofthefibroblaststoirradiation.AmericanJournalofPathology.2009:174(1):248-255.GiampietroPF,DunwoodieSL,KusumiK,Pourquie3.O,TassyO,OffiahAC,CornierAS,AlmanBA,BlankRD,RaggioCL,GlurichL,TurnpennyPD.Progressintheunderstandingofthegeneticetiologyofvertebralsegmentationdisordersinhumans.AnnalsNYAcademyofScience.2009:1151:38-67.LavoieJF,BiernaskieJA,ChenY,BagliD,AlmanB,4.KaplanDR,MillerFD.Skin-derivedprecursorsdifferentiateintoskeletogeniccelltypesandcontributetobonerepair.StemCellsDevelopment.2009:18(6):893-906.HoL,AlmanB.Protectingthehedgerow:p53and5.hedgehogpathway.CellCycle.2010:9(3):506-511.LipaJE,QiuW,HuangN,AlmanBA,PangCY.6.Pathogenesisofradiation-inducedcapsularcontractureintissueexpanderandimplantbreastreconstruction.PlasticReconstructionSurgery.2010:125(2):437-445.SzauterKM,OrdasA,LaxerRM,PopeE,Wherrett7.D,AlmanB,MinkM,BoydCD,CsiszarK,HinekA.Anovelfibroticdisorderassociatedwithincreaseddermalfibroblastproliferationanddownregulationofgenesofthemicrofibrillarnetwork.BritishJournalofDermatology.2010:163(5):1102-1115.OffiahA,AlmanB,CorneirAS,GiampietroPF,TassyO,8.WadeA,TurnpennyPD,ICVAS(InternationalConsortiumforVertebralAnomaliesandScoliosis).Pilotassessmentofaradiologicclassificationsystemforsegmentationdefectsofthevertebrae.AmericanJournalofMedicalGeneticsPartA.2010:152A(6):1357-1371.BovéeJVMG,HogendoornPC,WunderJS,AlmanBA.9.Cartilagetumorsandbonedevelopment:Molecularpathologyandhintsintotherapeutictargets.NatureReviewsCancer.2010:10(7):481-488.AlmanBA,WedgeJ.RobertBruceSalter,C.C.,MD,10.FRCSC.Dec15,1924-May10,2010.JournalofChildren'sOrthopaedics.2010:4(4):275-276.Amini-NikS,KraemerD,CowanML,GunaratneK,11.NadesanP,AlmanBA,MillerJD.Ultrafastmid-IRlaserscalpel:Proteinsignalsofthefundamentallimitstominimallyinvasivesurgery.PLoSONE.2010:5(9):pii:e13053.WuC,AminiNikS,NadesanP,StanfordWL,AlmanBA.12.Aggressivefibromatosis(desmoidtumor)isderivedfrommesenchymalprogenitorcells.CancerResearch.2010:70(19):7690-7698.RockelJS,AlmanBA.Don'thedgeyourbets:Hedgehog13.signalingasacentralmediatorofenchondralbone

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Benjamin A. Alman

developmentandcartilagediseases.JournalofOrthopaedicResearch.2011:29(6):810-815.SonnadaraRR,VanVlietA,SafirO,AlmanB,FergusonP,14.KraemerW,ReznickR.OrthopaedicBootcamp:Examiningtheeffectivenessofanintensivesurgicalskillscourse.Surgery.2011:149(6):745-749.HongH,NadesanP,PoonR,AlmanBA.Testosterone15.regulatescellproliferationinaggressivefibromatosis(desmoidtumour).BritishJournalofCancer.2011:104(9):1452-1458.BielefeldKA,Amini-NikS,WhetstoneH,PoonR,Youn16.A,WangJ,AlmanBA.Fibronectinandbeta-cateninactinaregulatoryloopindermalfibroblaststomodulatecutaneoushealing.JournalofBiologicalChemistry.2011:286(31):27687-27697.HsuSC,ZhangX,YuC,LiZJ,WunderJS,HuiCC,Alman17.BA.Kif7promotesHedgehogsignalingingrowthplatechondrocytesbyrestrictingtheinhibitoryfunctionofSufu.Development.2011:138(17):3791-3801.Amini-NikS,GlancyD,BoimerC,WhetstoneH,Keller18.C,AlmanBA.Pax7expressingcellscontributetodermalwoundrepairtegulatingscarsizethroughabeta-cateninmediatedprocess.StemCells.2011:29(9):1371-1379.Robanus-MaandagE,BoschC,Amini-NikS,Knijnenburg19.J,SzuhaiK,CerveraP,PoonR,EcclesD,RadiceP,GiovanniniM,AlmanBA,TejparS,DevileeP,FoddeR.Familialadenomatouspolyposis-associateddesmoidsdisplaysignificantlymoregeneticchangesthansporadicdesmoids.PLoSONE.2011:6(9):e24354.AtesokKI,AlmanBA,SchemitschEH,PeyserA,Mankin20.H.Osteoidosteomaandosteoblastoma.JournaloftheAmericanAcademyofOrthopaedicSurgeons.2011:19(11):678-689.AlmanBA,KelleySP,NamD.Healthyself:Using21.endogenousregenerationtorepairbone.TissueEngineering.PartBRev.2011:17(6):431-436.FattahA,CypelT,DonnerEJ,WangF,AlmanBA,Zuker22.RM.Thefirstsuccessfullowerextremitytransplantation:6-yearfollow-upandimplicationsforcorticalplasticity.AmericanJournalofTransplant.2011:11(12):2762-2767.WangCY,WeiQ,HanI,SatoS,Ghanbari-AzarnierR,23.WhetstoneH,PoonR,HuJ,ZhengF,ZhangP,WangW,WunderJS,AlmanBA.Hedgehogandnotchsignalingregulateself-renewalofundifferentiatedpleomorphicsarcoma.CancerResearch.2012:72(4):1013-1022.ZolkipliZ,SherlockM,BiggarWD,TaylorG,Hutchison24.JS,PeliowskiA,AlmanBA,LingSC,TeinI.Abnormalfattyacidmetabolisminspinalmuscularatrophymaypredisposetoperioperativerisks.EuropeanJournalofPediatricNeurology.2012:16(5):549-553.SparrowDB,ChapmanG,SmithAJ,MattarMZ,Major25.JA,O'ReillyVC,SagaY,ZackaiEH,DormansJP,AlmanBA,McGregorL,KageyamaR,KusumiK,DunwoodieSL.Amechanismforgene-environmentinteractioninthe

etiologyofcongenitalscoliosis.Cell.2012:149(2):295-306.PoonR,HongH,WeiX,PanJ,AlmanBA.Ahigh26.throughputscreenidentifiesNefopamastargetingcellproliferationinβ-catenindrivenneoplasticandreactivefibroproliferativedisorders.PLoSONE.2012:7(5):e37940.McAdamLC,MayoAL,AlmanBA,BiggarWD.The27.Canadianexperiencewithlong-termdeflazacorttreatmentinDuchennemusculardystrophy.ActaMyol.2012:31(1):16-20.SonnadaraRR,GarbedianS,SafirO,NousiainenM,Alman28.B,FergusonP,KraemerW,ReznickR.OrthopaedicBootCampII:Examiningtheretentionratesofanintensivesurgicalskillscourse.Surgery.2012:151(6):803-807.NamD,MauE,WangY,WrightD,SilkstoneD,Whetstone29.H,WhyneC,AlmanB.T-lymphocytesenableosteoblastmaturationviaIL-17Fduringtheearlyphaseoffracturerepair.PLoSONE.2012:7(6):e40044.AliSA,AlmanB.RNAextractionfromhumanarticular30.cartilagebychondrocyteisolation.AnalyticalBiochemistry.2012:429(1):39-41.BlackKP,AlmanBA,LevineWN,NestlerSP,PinneySJ.31.OrthopaedicResidentEducation-It'sawholenewgame:

“IfI'mgoingtobeaspinesurgeon,whydoIneedtolearnhowtoreconstructananteriorcruciateligament?”AOAcriticalissues.JournalofBoneandJointSurgery(Am).2012:94(13).MaezawaY,BinnieM,LiC,ThornerP,HuiC-C,Alman32.B,TaketoMM,QuagginSE.AnewCredrivermouseline,Tcf21/Pod1-Cre,targetsmetanephricmesenchyme.PLoSONE.2012:7(7):e40547.TsoiKM,DaiQ,AlmanBA,ChanWC.Arequantumdots33.toxic?Exploringthediscrepancybetweencellcultureandanimalstudies.AccChemRes.2012:Aug1(Epubaheadofprint).BielefeldKA,Amini-NikS,AlmanBA.Cutaneous34.woundhealing:Recruitingdevelopmentalpathwaysforregeneration.CellularandMolecularLifeSciences.2012:Oct4(Epubaheadofprint).HsuC,ZhangX,ChengS,WunderJS,HuiCC,Alman35.BA.Suppressoroffused(Sufu)mediatestheeffectofParathyroidhormone-likehormone(Pthlh)onchondrocytedifferentiationinthegrowthplate.JournalofBiologicalChemistry.2012:287(43):36222-36228.HoL,AliSA,Al-JazraweM,KandelR,WunderJS,36.AlmanBA.PrimaryciliaattenuateHedgehogsignallinginneoplasticchondrocytes.Oncogene.2012:Dec17(Epubaheadofprint)..CampM,MattinglyDA,GrossAE,NousiainenMT,Alman37.BA,McKneallyMF.Patients'viesonsurgeons'financialconflictsofinterest.JournalofBoneandJointSurgery(Am).2013:95(2):e91-98.AlmanBA,FergusonP,KraemerW,NousiainenMT,38.ReznickRK.Competency-basededucation:anewmodelforteachingorthopaedics.InstrCourseLect.2013;62:565-9.

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Currenttherapeuticstrategiesofcatabolicbonediseasessuchasosteoporosisandmetastatictumorsinbonetargetosteoclastsinordertoreduceornormalizetheelevatedboneresorptionactivity.BothbisphosphonatesandhumanizedmonoclonalantibodyagainstRANKLaresuchanti-resorptiveagentsachievingthistherapeuticgoalalbeitthroughdifferentpharmacokinetics.

Osteonecrosisofthejaw(ONJ)inmaxillaandmandiblehasemerged,however,asararebutseveresideeffectreportedbetween0.5%to18%amongthosepatientswhoweretreatedwithanti-resoptiveagents.Whileclinicalpresentationsvarysignificantly,ONJsymptomscommonlydemonstratethelackofresolutioninchronicinflammationoforalmucosa.Oralmucosaiscomposedofstratifiedepitheliumandthinconnectivetissue,andconsideredoneofthemostprotectivetissuebarriersfromenvironmentalstresses,chemicaldamageandbacterialinfection.BarriertissuesareknowntocontainTcellsexpressingacanonicalgammadeltaTcellreceptor.ImmunosurveillanceandpromptresponsetoinjurybythebarriertissuearethoughttoinvolvegammadeltaTcells.

OurrecentstudiesusingTcrd-H2BEGFPmicedemonstratedtheprolongedretentionofgammadeltaTcellsingingival/palataltissueaftermaxillarymolarextractionwhenmicewereinjectedwithzoledronate(ZOL).Furthermore,thedevelopmentofONJ-likelesionwassignificantlymodulatedinZOL-treatedTcrd-/-mice.Thus,wepostulatethatthecloseapproximationbetweenjawboneandoralbarrierimmunitymay,inpart,contributetothepathogenesisofONJ.

ItwasnotedthatanimalmodelsofONJ-likelesionrevealedanunusualco-localizationofinflammatorycellswithZOL-affectedosteoclasts.OsteoclastsaredifferentiatedfromthemonocyticlineageofmyeloidimmunecellswiththepresenceofM-CSFandRANKL.Oncedifferentiated,osteoclastshavebeenconsideredtofunctionasbonecells.However,theuniqueandconsistentobservationonthedistinctassociationbetweenZOL-affectedosteoclastsandlymphocytesaswellasneutrophilsmaysuggestayetuncoveredfunctionofosteoclastsasimmunecells.

OurpreliminarycharacterizationofosteoclastsderivedfromhumanCD14+monocytessuggestedthesecretionofasetofcytokines.Typically,osteoclasticcytokinesexhibitedasimilarprofileofmyeloid-derivedsuppressorcells(MDSC).MDSCnotonlysuppresseslymphocytescontributingtotheresolutionofinflammation;butfurthersupportstissuerepairbyinducingvascularformationandsecretinggrowthfactors.Thispresentationwillhighlightanewconceptofosteoclastactivitiesasanimmunecells,inparticular,withMDSC-likecharacteristics.Contributionsofosteoclaststophysiologicalboneremodelingandcouplingwithosteoblasts,aswellaspathologicalcontributionstoONJandmetastatictumorsinbonemarrowwillbediscussed.

Osteoclast as MDSC (myeloid-derived suppressor cell) and its role inONJ and tumor bone metastasis

Ichiro Nishimura, D.D.S., D.M.Sc., D.M.D., F.A.P.

ProfessorWeintraubCenterforReconstructiveBiotechnologyUCLASchoolofDentistry

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Ichiro Nishimura

CURRICULUM VITAE

Education

1981 D.D.S.TokyoDentalCollege,Tokyo,Japan1986 D.M.Sc.HarvardUniversity,Cambridge, MA(Ph.D.equivalent)1986 CertificatePostgraduateProsthodontics(BoardEligible) HarvardSchoolofDentalMedicine,Boston,MA1993 D.M.D.HarvardSchoolofDentalMedicine,Boston,MA

Position

1990-93 FacultyMember,CommitteeofCell& DevelopmentalBiologyHarvardMedicalSchool, Boston,MA1994-97 AssociateProfessorofProstheticDentistry HarvardSchoolofDentalMedicine,Boston,MA1997-99 AssociateProfessor(withtenure) AdvancedProsthodontics,Biomaterialsand HospitalDentistry UCLASchoolofDentistry,LosAngeles,CA1999-PresentProfessor(withtenure) AdvancedProsthodontics,Biomaterialsand HospitalDentistry UCLASchoolofDentistry,LosAngeles,CA2000-PresentProfessorofOralBiology(JointAppointment) UCLASchoolofDentistry,LosAngeles,CA2012-PresentAffiliateProfessorofBioengineering DepartmentofBioengineering UCLAHenrySamueliSchoolofEngineeringand AppliedScience

Award and Honors

2000 FellowofBiotechnology:PolicyIssuesandRegulatory FrameworksSalzburgSeminar,Salzburg,Austria2000 AppreciationofServiceU.S.DepartmentofHealthand HumanServices,PublicHealthService,NIH,National InstituteofDentalCraniofacialResearch2004 DistinguishedScientistAwardinProsthodonticsand ImplantologyInternationalAssociationforDental Research2004 TheodoreM.HesburghCertificateofExcellencefor FacultyDevelopmenttoEnhanceUndergraduate TeachingandLearningUCLAFreshmanCluster ProgramFacultymemberinBiotechnologyandSociety TIAA-CREF2005 AppreciationofServiceU.S.DepartmentofHealthand HumanServices,PublicHealthService,NIH,National InstituteofDentalCraniofacialResearch

Publications

YukiI、LeeD,MurayamaY,ChiangA,VintersHV,1.NishimuraI,WangJCC,IshiiA,BenjaminWM,VinuelaF:Thrombosisandhealingintheexperimentalaneurysmmodel,partII:Thrombusorganizationinthesettingofbioabsorbablepolymers,JNeurosurg ,107:109-120,2007.NishimuraI:Nanotechnologyanditsapplicationsindental2.implant,ClinResDent ,4:29-35,2007.ThakorD,SpigelmanI,TabataY,NishimuraI.:3.Subcutaneousperipheralinjectionofcationizedgelatin/DNApolyplexesasaplatformfornoninvasivegenetransfertosensoryneurons,MolecularTherapy ,15:2124-2131,2007.EgusaH,IidaK,KobayashiM,LinT,ZhuM,ZukP,Hedrik4.M,NishimuraI:Down-regulationofextracellularmatrix-relatedgeneclusterduringosteogenicdifferentiationofbonemarrowandadiposetissue-derivedstromalcells.TissueEng ,13:2589-2600,2007.OhJ,WangCJ,PooleM,KimE,DavisRC,NishimuraI,5.PaeEK:Thegenomesegmentonmousechromosome12determinesmaxilla,JDentRes ,86:1203-1206,2007.WangCJ,IidaK,EgusaH,HokugoA,JewettA,Nishimura6.I:Trabecularbonedeteriorationincol9a1+/- miceassociatedwithenlargedosteoclastsadheredtocollagenIX-deficientbone,JBoneMinerRes ,23(6):837-849,2008.OhJ-H,NishimuraI.:Copynumbervariationinthehuman7.genomeanditsimplicationforhumandisease.JKoreanAcadMaxillofacPlasticReconstrucSurg ,30:205-212,2008.ThakorDK,LinA,MatsukaY,MeyerE,RuangsriS,8.NishimuraI,SpigelmanI:IncreasedperipheralnerveexcitabilityandlocalNaV1.8mRNAup-regulationinpainfulneuropathy,MolPain ,5(1):14,2009.[Openaccess][Highlyaccessedarticle ]KellyJ,LinA,WangCJ,ParkS,NishimuraI:Theeffect9.ofvitaminDinsufficiencyonimplantosseointegration.JProsthodont ,18(6):473-478,2009.LinA,WangCJ,KellyJ,GubbiP,NishimuraI:Theroleof10.titaniumimplantsurfacemodificationwithhydroxyapatititenanoparticlesinprogressiveearlybone-implantfixationinvivo .IntJOralMaxiofacImplants ,24(5):808-816,2009.NishimuraI,ThakorD,LinA,RuangsriS,Spigelman11.I:Molecularstrategiesfortherapeutictargetingofprimarysensoryneuronsinchronicpainsyndromes,InTranslationalPainResearch:FromMousetoMan ,Ed.KrugerL,LightA.pp139-161,CRCPress,BocaRaton,FL,2009.LinA,HokugoA,ChoiJ,NishimuraI:Smallcoiled-coil12.cytoskeleton-associatedmolecule,FGFR1OP2/wit3.0facilitatesfibroblast-drivenwoundclosure.AmJPathol ,176(1):108-121,2010.HokugoA,ChristensenR,ChungEM,SungEC,Felsenfeld13.AL,SayerJ,GarrettN,AdamsJS,NishimuraI:Increasedprevalenceofbisphosphonate-relatedosteonecrosisofthejawwithvitaminDdeficiencyinrats.JBoneMinRes ,

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Ichiro Nishimura

25(6):1337-49,2010.Nishimura,I:Implantinterfacetobonetissue:Biomimetic14.surfacefunctionalizationthroughnanotechnology,InInterfaceOralHealthScience2009 ,Ed,FukumotoT,IwamoriM,SuzukiO,pp60-66,SpringerJapan,Tokyo,Japan,2010.YukiI,UchiyamaN,MurayamaY,NienYL,LeeD,15.EbaraM,IshiiA,ChiangA,VintersHV,NishimuraI,WuBM,VinuelaF:Intravasculartissuereactionsinducedbyvarioustypesofbioabsorbablepolymericmaterials:correlationbetweenthedegradationprofilesandcorrespondingtissuereactions.Neuroradiology ,52(11):1017-24,2010.LinA,HokugoA,NishimuraI:Woundclosureandwound16.management:Anewtherapeuticmoleculartarget.CellAdhesion&Migration ,4(3):396-399,2010.MussanoF,LeeK,ZukP,TranL,CacalanoN,Jewett17.A,CarossaS,NishimuraI:Differentialeffectofionizingradiationexposureonmultipotentanddifferentiation-restrictedbonemarrowmesenchymalstemcells.JCellBiochem,111(2):322-32,2010.NishimuraI.:GEM21S:Futureperspectivesofdental18.biologics(commissionedreview:inJapanese).DentalDiamond ,2010.CohenM,NishimuraI,TamplenM,HokugoA,Beumer19.J,SteinbergM,ShuJD,AbemayerE,NabiliV:Animalmodelofradiogenicbonedamagetostudymandibularosteoradionecrosis,AmJOtolaryngol ,32(4):291-300,2011.MengattoCM,MussanoF,HondaY,ColwellC,NishimuraI:20.Circadianrhythmandcartilageextracellularmatrixgenesinosseointegration:Agenome-widescreeningofimplantfailurebyvitaminDdeficiency.PloSOne ,6(1):e15848,2011.SuwanwelaJ,LeeJ,LinA,UcerC,DevlinH,Sinsheimer21.J,GarrettN,NishimuraI:AGeneticAssociationStudyofSingleNucleotidePolymorphismsinFGFR1OP2/wit3.0andLong-termAtrophyofEdentulousMandible.PloSOne ,6(1):e16204,2011.TempletonM,TrappK,NishimuraI,ArminB,SteinbergM,22.BeumerJ,AbemayorE,NabiliV:Standardizedanalysisofmandibularosteoradionecrosisinaratmodel,OtolaryngolHeadNeckSurg ,145(3):404-10,2011.ButzF,OgawaT,NishimuraI:Interfacialshearstrength23.ofendosseousimplants,IntJOralMaxillofacImp ,26(4):746-51,2011.MawardiH,GiroG,KajiyaM,OhtaK,AlmazrooaS,24.AlshwaimiE,WooS-B,NishimuraI,KawaiT:Aroleoforalbacteriainbisphosphonate-inducedosteonecrosisofjaw,JDentRes ,90(11):1339-45,2011.RuangsriS,LinA,MulpuriY,LeeK,SpigelmanI,25.NishimuraI:AxonalNav1.8mRNAaccumulationandsciaticnerveinjury-inducedpainfulneuropathyinrats,JBiolChem,286(46):39836-47,2011.HondaY,DingX,MussanoF,WibergA,HoCM,26.NishimuraI:Applicationoffeedbacksystemcontrol(FSC)

toidentifytheoptimizedosteogenicdrugcocktails,IEEE,10.1109/MHS.2011.6102147,2011.ArminBB,HokugoA,NishimuraI,TamplenM,Beumer27.JIII,SteinbergML,AbemayorE,NabiliV.Brachytherapy-mediatedbonedamageinaratmodelinvestigatingmaxillaryosteoradionecrosis,ArchOtolaryngolHeadNeckSurg ,138(2):167-171,2012.KahenasaN,SungE,NabiliV,KellyJ,GarretN,Nishimura28.I:Resolutionofpainandcompletehealingofmandibularosteoradionecrosisusingpentoxifyllineandtocopherol,acasereport,OralSurgOralMedOralPatholOralRadiolEndodontol ,113(4):e18-e23,2012.ChengW,NishimuraI:HighThroughputScreeningof29.BiologicallyFunctionalSmallMoleculesforModulatingtheExpressionofFGFR1OP2/wit3.0inFibroblasts,JCalDentAssoc ,40(12):929-31,934-7,2012.HokugoA,SunS,ParkS,McKennaC,NishimuraI:30.Equilibrium-dependentbisphosphonateinteractionwithcrystallinebonemineralexplainstheanti-resorptivepharmacokineticsandtheprevalenceofosteonecrosisofthejawinaratmodel,Bone ,53(1):59-68,2013.HaJ,OsherSJ,NishimuraI:Mathematicalfiltering31.minimizedmetallichalationoftitaniumimplantinmicroCTimage,JCalDentAssoc ,41(1):41-5,2013.DevlinH,NishimuraI(Editors):OralandCranialImplants32.–RecentResearchDevelopment,Springer,Heidelberg,Germany,2013.NishimuraI:Geneticbackgroundofimplantfailure,in33. OralandCranialImplants–RecentResearchDevelopments ,DevlinH,NishimuraI,Eds.Springer,Heidelberg,Germany,2013.ParkS,NishimuraI:Biosphosphonatesanddentalimplant34.treatment,inOralandCranialImplants–RecentResearchDevelopments ,DevlinH,NishimuraI,Eds.Springer,Heidelberg,Germany,2013.NishimuraI:Geneticnetworksinosseointegration,35. JDentRes ,inpress,2013.HondaY,DingX,MussanoF,WibergA,HoCM,36.NishimuraI:Guidingosteogenicfateofmouseandhumanmesenchymalstemcellsthroughfeedbacksystemcontrol,ScientificReports ,Accepted,2013.KellyJA,MallyaS,NabiliV,NishimuraI:Currentclinical37.managementofintraoralosteoradionecrosis,JProsthetDent ,Submitted,2013.NishimuraI:Acquisitionandmaintenanceof38.osseointegration,InImplantSurgery ,BeumerJIIIEd.Quintessence,Chicago,IL,Submitted,2013.NishimuraI,GarrettN,BeumerJ:Edentulouspatients,39.Patternsofboneresorptionandclinicaloutcomes:osseointegratedimplants,InImplantSurgery ,BeumerJIIIEd.Quintessence,Chicago,IL,Submitted,2013.

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P o s t e r S e s s i o n

Retreat 2014

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Poster Session List

Name Grade

MaChengshan 4

NurmaaDashzeveg 4

BijayaHaobam 4

RumanaKhanom 4

DuartePuertoCarolinaLizeth 4

ZayarLin 4

JunpeiShirakawa 4

YukihikoHashida 4

Md.SofiqulIslam 4

MayumiOgita 4

NadilaWali 4

TadashiHosoya 4

NaokiKimura 4

WaruneePluemsakunthai 4

ChengXu 4

GerardoJoseJovesMendez 4

MasayoshiUezono 4

DawudAbduweli 3

GuJie 3

SuphanantachatSupreda 3

LiHui 3

SurapornsawasdThunyaporn 3

ThanitPrasitsak 3

RajapaksheMudiyanselageAnupamaRasadariRajapakshe 3

ThanatvarakornOrnnicha 3

YusukeMatsuo 3

YokoYoshihashi 3

MaheswariKuppusamy 3

MohannadIssaMichaelNassar 3

KenjiOgura 3

KahaerAbula 3

TakayukiYamada 3

UeharaDanielaTiaki 2

Nuylan,MichelleLoyola 2

AlaaAbdulahadTurkistani 2

ALSAYED,EHABZAKIE 2

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Name Grade

HuaQiao 2

Khunkar,SaharJameelM 2

KenchiTakenaka 2

KongKalyan 2

BabaBista 2

MakiriKawasaki 2

NatsukaUmezawa 1

JunYamada 4

TakashiTaniyama 4

Munetaka　Iwata 4

ArataYuki 4

MadokaOnuma 4

NobutakeOzeki 4

YuMatsukura 3

YusukeNakagawa 3

HidetoshiKaburagi 2

MasanoriSaito 2

SatoshiSumiya 2

KatsuakiYanagisawa 2

MakikoInoue 2

MioUdo 2

ShinpeiKondo 2

RyusukeSaito 2

ToshiyukiOhara 2

MikioShioda 1

KaoriNakamura 1

HiroakiYasuda 1

JinyingPiao

KojiFujita

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ゴミ箱

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