神经退行性疾病干细胞移植治疗目前研究现状与未来展望培训讲义课件

神神经经退行性疾病干退行性疾病干细细胞移植治胞移植治疗疗目前研究目前研究现现状与未来展状与未来展望望神神经经退行性疾病干退行性疾病干细细胞移植治胞移植治疗疗目前研究目前研究现现状与未来展望状与未来展望1（优选）神经退行性疾病干细胞移植治疗目前研究现状与未来展望（优选优选）神）神经经退行性疾病干退行性疾病干细细胞移植治胞移植治疗疗目前研究目前研究现现状与未来展望状与未来展望胎儿神经干细胞治疗帕金森氏病临床研究发展历程胎儿神经干细胞治疗帕金森氏病临床研究发展历程EvansJR,MasonSL,BarkerRA.ProgBrainRes.2012;200:169-98胎儿神胎儿神经经干干细细胞治胞治疗疗帕金森氏病帕金森氏病临临床研究床研究发发展展历历程程EvansJRLindvallO,et al，NatMed.2008May;14(5):501-3THSynucleinOverlayTransplanted fetal mesencephalic dopaminergic neurons(11-16 years)developed alpha-synuclein-positive Lewy bodies in grafted neuronsLindvallO,etal，NatMed.200Synuclein-HostUbiquintin-HostSynuclein-GraftedNeuronsUbiquintin-GraftedNeuronsGrafted nigral neurons were found to have Lewy body-like inclusions14 years after transplantation into the striatum of an individual with PDOlanowCW.etalNatMed.2008May;14(5):504-6.Synuclein-HostUbiquintin-HostSTransplanted dopamine neurons in people with PD do not contain Lewy bodiesMendez,Isacsonetal,NATUREMEDICINEVOLUME14(5):507-509,2008TransplanteddopamineneuronsFreed CR,JNuclMed.2010Jan;51(1):7-15-LongtermStudy-33oftheoriginaltrialparticipantswhowerefollowedfor2yearsaftertransplantationand15ofthesesubjectswhowerefollowedfor2additionalyears.-Theseresultssuggestthatclinicalbenefitandgraftviabilityaresustainedupto4yaftertransplantation.FreedCR,Neurotherapeutics(2011)8:549561FreedCR,JNuclMed.2010Jan人体胚胎干细胞分化的多巴胺神经元移植人体胚胎干细胞分化的多巴胺神经元移植改善小鼠，大鼠和猴子帕金森氏病的运动障碍改善小鼠，大鼠和猴子帕金森氏病的运动障碍22/29 DECEMBER 2011|VOL 480|NATURE|547，Lorenz Studer，et al Memorial Sloan-Kettering Cancer Center人体胚胎干人体胚胎干细细胞分化的多巴胺神胞分化的多巴胺神经经元移植元移植改善小鼠，大鼠和猴子帕改善小鼠，大鼠和猴子帕ImprovedCellTherapyProtocolforParkinsonsDiseaseBasedonDifferentiationEfficiencyandSafetyofhESC-,HipscandNon-HumanPrimateiPSC-DerivedDANeuronsIsacsonetal,StemCells.2013;31(8):1548-62.ImprovedCellTherapyProtocolDopaminereleasefromtransplantedneuralstemcellsinParkinsonianratstriatuminvivo.Twelvepatientsweretreatedwitheitherunilateralorbilateralinjections.6OHDAinducedRatPDModel分化的胎脑神经干细胞移植治疗PDIncreasinghostneurogenesis“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.TodeterminewhetherlocalmechanismsmediateBMSCneuroprotectiveactionsetalNatMed.（优选）神经退行性疾病干细胞移植治疗目前研究现状与未来展望胎儿神经干细胞治疗帕金森氏病临床研究发展历程Releaseofimmuno-regulatoryfactorssuchasIL-2,6,8,10toplayimmuno-modulationandattenuationoftheinflammatoryprocess,suchasMSC.2012;200:169-98ImprovedCellTherapyProtocolforParkinsonsDiseaseBasedonDifferentiationEfficiencyandSafetyofhESC-,HipscandNon-HumanPrimateiPSC-DerivedDANeuronsIntraspinalstemcelltransplantationinALS:aphaseItrial,2014Anotherrequiredreoperationforwounddehiscenceandinfection.usingESCsandiPSCsiPSCellsWereGeneratedfromPDpatientsandNormalControlsUbiquintin-GraftedNeuronsIncreasinghostneurogenesisgraftedallogeneicBMSCstositesofsevere,compressivespinalcordinjury(SCI)inSpragueDawleyrats.Dopamine release from transplanted neural stem cells in Parkinsonian rat striatum in vivo.Zhouz,etal,ProcNatlAcadSciUSA.2014Nov4;111(44):15804-9DopaminereleasefromtransplaiPSCDerived Dopamine Neurons function after Transplantation in a NonHuman Primate Model of Parkinsons DiseaseCellStemCell.2015Mar5;16(3):269-74.OleIsacsonetal，HarvardStemCellInstituteiPSCDerivedDopamineNeuronsfStem cellbased Clinical Trials for(ALS)Nuralstem,Inc.thefirstPhaseIclinicaltrialforastemcell-basedtreatmentofALS.Initiatedin2010andcompletedin2013,involvedthetransplan-tationofhumanspinalcord-derivedNSCsintothespinalcordof15latetomid-stageALSpatientsStemcellbasedClinicalTrials“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.SummaryonMolecularMechanismofStemCellTransplantationforNeurologicalDiseasestoincreasethefunctionsoftheendogenousneuralstemcellsThisencouragingfindingsupportsconsiderationofthisdeliveryapproachforneurodegenerative,oncologic,andtraumaticspinalcordafflictions.暴露和部分横切脊髓外科手术。University,Atlanta,Georgia;DepartmentofNeurology,EmoryUniversity,Atlanta,Georgia;DepartmentofNeurology,UniversityofMichigan,AnnArbor,MichiganGlass,Feldman,E.RTPCRtoDetecttheMicroRNAExpressioninRatSCIModelPatientsarefollowedclinicallyandradiologicallytoassesspotentialtoxicityoftheprocedure.Cellswereadministered48hoursaftertheoriginalinjury.SCgraftsshouldexhibitregulatedreleaseofdopamineinlinewiththatofendogenousdopaminergicneurons.-LongtermStudy神经退行性疾病干细胞移植治疗目前研究现状与未来展望Isacsonetal,StemCells.神经退行性疾病干细胞移植治疗目前研究现状与未来展望RealTimeRTPCRtoDetecttheMicroRNAExpressioninSCIiPSCellsWereGeneratedfromPDpatientsandNormalControlsGlass,Feldman,E.Thus,allogeneicgraftsofBMSCsdonotappeartoactthroughlocalmechanisms,andfutureclinicaltrialsthatacutelydeliverBMSCstoactualsitesofinjurywithindaysareunlikelytobebeneficial.Ubiquintin-GraftedNeurons“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.Thisencouragingfindingsupportsconsiderationofthisdeliveryapproachforneurodegenerative,oncologic,andtraumaticspinalcordafflictions.IntraspinalstemcelltransplantationinALS:aphaseItrial,2014Glass,Feldman,E.L.,2012.Lumbarintraspinalinjectionofneuralstemcellsinpatientswithamyotrophiclateralsclerosis:resultsofaphaseItrialin12patients.StemCells30(6),11441151.Riley,J.,Feldman,E.L.,2014.“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.Neurosurgery74(1),7787“IntraspinalstemcelltransplRESULTS:Unilateralcervical(groupD,n=3)andcervicalplusthoracolumbar(groupE,n=3)microinjectionstotheventralhornhavebeencompletedinambulatorypatients.Onepatientdevelopedapostoperativekyphoticdeformitypromptingcompletionofalaminoplastyinsubsequentpatients.Anotherrequiredreoperationforwounddehiscenceandinfection.Thesolitarypatientwithbulbaramyotrophiclateralsclerosisrequiredperioperativereintubation.CONCLUSION:Deliveryofacellularpayloadtothecervicalorthoracolumbarspinalcordwaswelltoleratedbythespinalcordinthisvulnerablepopulation.Thisencouragingfindingsupportsconsiderationofthisdeliveryapproachforneurodegenerative,oncologic,andtraumaticspinalcordafflictions.IntraspinalstemcelltransplantationinALS:aphaseItrial,2014RESULTS:Unilateralcervical(iPS Cells Were Generated from PD patients and Normal Controls iPSCellsWereGeneratedfrom6OHDAinduced Rat PD Model6OHDAinducedRatPDModelHuman iPS cells Integrated to the Host Brain of 6OHDAinduced Rat PD ModelHanF,WangW,ChenC,DuanJ,etalCytotherapy2015HumaniPScellsIntegratedto分化的胎脑神经干细胞移植治疗分化的胎脑神经干细胞移植治疗PD分化的胎分化的胎脑脑神神经经干干细细胞移植治胞移植治疗疗PD建立大鼠建立大鼠SCISCI损伤模型损伤模型A.A.暴露和暴露和部分部分横切脊髓外科手术。横切脊髓外科手术。B.T7 B.T7 横断损伤产生后肢横断损伤产生后肢瘫痪。瘫痪。C.C.无脊髓损伤的正常大鼠。无脊髓损伤的正常大鼠。建立大鼠建立大鼠SCI损伤损伤模型模型A.暴露和部分横切脊髓外科手暴露和部分横切脊髓外科手术术。BRTPCR to Detect the MicroRNA Expression in Rat SCI Model MiR-124MiR-124MiR-124MiR-127MiR-127MiR-127MiR-127MiR-124MiR-133aMiR-133aMiR-133aMiR-181aMiR-181aMiR-181aRTPCRtoDetecttheMicroRNAERealTime RTPCR to Detect the MicroRNA Expression in SCIRealTimeRTPCRtoDetecttheM干细胞移植修复脊髓神经损伤干细胞移植修复脊髓神经损伤干干细细胞移植修复脊髓神胞移植修复脊髓神经损伤经损伤移植神经干细胞分化的神经轴索与宿主脊髓神经细移植神经干细胞分化的神经轴索与宿主脊髓神经细胞及其树突形成突触连接胞及其树突形成突触连接 Lu P et al Lu P et al，Cell.2012 September 14;150(6):12641273Cell.2012 September 14;150(6):12641273移植神移植神经经干干细细胞分化的神胞分化的神经轴经轴索与宿主脊髓神索与宿主脊髓神经细经细胞及其胞及其树树突形成突突形成突BoneMarrowStromalCellIntraspinalTransplantsFailtoImproveMotorOutcomesinaSevereModelofSCIJournalofNeurotrauma2015，TuszynskiMHTodeterminewhetherlocalmechanismsmediateBMSCneuroprotectiveactionsgraftedallogeneicBMSCstositesofsevere,compressivespinalcordinjury(SCI)inSpragueDawleyrats.Cellswereadministered48hoursaftertheoriginalinjury.AdditionalanimalsreceivedallogeneicMSCsthatweregeneticallymodifiedtosecreteBDNF,tofurtherdeterminewhetheralocallyadministeredneurotrophicfactorprovidesorextendsneuroprotection.twomonthspost-injuryinaclinicallyrelevantmodelofsevereSCI,BMSCgraftswithorwithoutBDNFsecretionfailedtoimprovemotoroutcomes.Thus,allogeneicgraftsofBMSCsdonotappeartoactthroughlocalmechanisms,andfutureclinicaltrialsthatacutelydeliverBMSCstoactualsitesofinjurywithindaysareunlikelytobebeneficial.BoneMarrowStromalCellIntraIntraspinalStemCellTransplantationinAmyotrophicLateralSclerosis:APhaseISafetyTrial,TechnicalNote,andLumbarSafetyOutcomesNEUROSURGERYVOLUME71|NUMBER2|AUGUST2012DepartmentofNeurosurgery,EmoryUniversity,Atlanta,Georgia;DepartmentofNeurology,EmoryUniversity,Atlanta,Georgia;DepartmentofNeurology,UniversityofMichigan,AnnArbor,MichiganIntraspinalStemCellTranspla神经干细胞移植方法神经干细胞移植方法Eachmicroinjectionseriescomprised5injections(10mL/injection)separatedby4mm.Eachinjection:100000neuralstemcellsderivedfromafetalspinalcord.Twelvepatientsweretreatedwitheitherunilateralorbilateralinjections.Patientsarefollowedclinicallyandradiologicallytoassesspotentialtoxicityoftheprocedure.神神经经干干细细胞移植方法胞移植方法EachmicroinjectionLumbarLaminectomyLumbarLaminectomyMicroinjection platform applicationMicroinjectionplatformapplicPostoperative imaging progressionPostoperativeimagingprogressRiley,J.,Feldman,E.L.,2014.“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.Neurosurgery 74(1),77 87Riley,J.,Feldman,E.L.,2014 Clinical Trials using ESCs and iPSCsClinicalTrialsThereisalsoareportofoneJapanesepatientwhoreceivedatransplantofasheetofiPSC-derivedRPEThereisalsoareportofoneSummary on Molecular Mechanism of Stem Cell Transplantation for Neurological DiseasesTransplanted cells survive，differentiate to neurons,astrocytes,oligodendrocyte precursors(hESC,hiPSC,NSC)and release neurological transmittors such as dopamine,Ach.Release of neurotrophic factors(GDNF,GDNE,IGF,)to increase the functions of the endogenous neural stem cellsRelease of immuno-regulatory factors such as IL-2,6,8,10 to play immuno-modulation and attenuation of the inflammatory process,such as MSC.The transplanted cells formed synapse with host cells.Others such as delaying the onset and prolonging survival of SOD1 rats Increasing host neurogenesis SummaryonMolecularMechanismOleIsacsonetal，HarvardStemCellInstituteEachinjection:100000neuralstemcellsderivedfromafetalspinalcord.Cellswereadministered48hoursaftertheoriginalinjury.sheetofiPSC-derivedRPEOtherssuchasdelayingtheonsetandprolongingsurvivalofSOD1ratsEachmicroinjectionseriescomprised5injections(10mL/injection)separatedby4mm.“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.SummaryonMolecularMechanismofStemCellTransplantationforNeurologicalDiseasesIntraspinalstemcelltransplantationinALS:aphaseItrial,2014Glass,Feldman,E.University,Atlanta,Georgia;DepartmentofNeurology,EmoryUniversity,Atlanta,Georgia;DepartmentofNeurology,UniversityofMichigan,AnnArbor,MichiganAmyotrophicLateralSclerosis:Cellswereadministered48hoursaftertheoriginalinjury.ThereisalsoareportofoneJapanesepatientwhoreceivedatransplantofa,Feldman,E.AmyotrophicLateralSclerosis:2008May;14(5):504-6.“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.Twelvepatientsweretreatedwitheitherunilateralorbilateralinjections.Mendez,Isacsonetal,NATUREMEDICINEVOLUME14(5):507-509,2008干细胞移植修复脊髓神经损伤AmyotrophicLateralSclerosis:OleIsacsonetal，HarvardStemCellInstituteIntraspinalstemcelltransplantationinALS:aphaseItrial,2014“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.神经退行性疾病干细胞移植治疗目前研究现状与未来展望Patientsarefollowedclinicallyandradiologicallytoassesspotentialtoxicityoftheprocedure.OleIsacsonetal，HarvardStemCellInstituteThisincludestheabsenceoftumorformationandGIDs（graft-induceddyskinesia）throughoutlong-termfollow-upperiods.2012September14;150(6):12641273Ubiquintin-GraftedNeuronsIncreasinghostneurogenesisEvaluationSystem（Symptoms,Dopaminerelease,Levodopa-response),UnifiedParkinsonsDiseaseRatingScale(UPDRS)，BiomarkersTrial,TechnicalNote,andLumbarSafetyOutcomes“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.,Feldman,E.Adverseeffectsmustbeminimal.GraftednigralneuronswerefoundtohaveLewybody-likeinclusions14yearsaftertransplantationintothestriatumofanindividualwithPD“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.DopaminereleasefromtransplantedneuralstemcellsinParkinsonianratstriatuminvivo.分化的胎脑神经干细胞移植治疗PDiPSCellsWereGeneratedfromPDpatientsandNormalControlstoincreasethefunctionsoftheendogenousneuralstemcells6OHDAinducedRatPDModel2010Jan;51(1):7-15“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.ThereisalsoareportofoneJapanesepatientwhoreceivedatransplantofa-LongtermStudyThereisalsoareportofoneJapanesepatientwhoreceivedatransplantofaPatientsarefollowedclinicallyandradiologicallytoassesspotentialtoxicityoftheprocedure.Patientsarefollowedclinicallyandradiologicallytoassesspotentialtoxicityoftheprocedure.SCgraftsshouldexhibitregulatedreleaseofdopamineinlinewiththatofendogenousdopaminergicneurons.APhaseISafety暴露和部分横切脊髓外科手术。“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.Zhouz,etal,ProcNatlAcadSciUSA.IntraspinalStemCellTransplantationinCellSources：Neuralprojenitors，MSC，hEScells，iPScellsSCgraftsshouldexhibitregulatedreleaseofdopamineinlinewiththatofendogenousdopaminergicneurons.LorenzStuder，etalMemorialSloan-KetteringCancerCenter2012;200:169-98-LongtermStudyCellswereadministered48hoursaftertheoriginalinjury.今后干细胞治疗神经退行性疾病的临床研究今后干细胞治疗神经退行性疾病的临床研究需要考虑的问题需要考虑的问题1.Cell Sources：Neural projenitors，MSC，hES cells，iPS cells 2.SC grafting should be conducted to ensure 100,000 dopaminergic neurons（PD）survive per transplantation site.3.SC grafts should exhibit regulated release of dopamine in line with that of endogenous dopaminergic neurons.4.By reestablishing the striatal dopaminergic system,grafts should show the capacity to restore functional connectivity within the basal ganglia and at extra-striatal loci.5.Long-lasting and significant symptom-relief must be achieved （over 2-3 years）.6.Evaluation System（Symptoms,Dopamine release,Levodopa-response),Unified Parkinson s Disea se Ra ting Scal e(UPDRS)，Biomarkers7.Adverse effects must be minimal.This include s the absence of tumor formation and GIDs（graf t-induced dyskinesia）throughout long-term follow-up periods.8.Sample Size：over 50-100 patients。OleIsacsonetal，HarvardStem