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 PPT课件.ppt

韩发彬，韩发彬，MD，PhD泰山医学院聊城临床学院泰山医学院聊城临床学院干细胞与再生医学实验室干细胞与再生医学实验室2015，10，31神经退行性疾病神经退行性疾病干细胞移植干细胞移植治疗：治疗：目前目前研究研究现状与未来展望现状与未来展望Stem Cell Sources for Treatment of Neurological Diseases(AD,PD,ALS,MS,Stroke,SCI)FabinHan,etal,JournalofNeurorestoratology2015:3112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.Prog Brain Res.2012;200:169-98Lindvall O,et al，Nat Med.2008May;14(5):501-3THSynucleinOverlayTransplanted fetal mesencephalic dopaminergic neurons(11-16 years)developed alpha-synuclein-positive Lewy bodies in grafted neuronsSynuclein-HostUbiquintin-HostSynuclein-GraftedNeuronsUbiquintin-GraftedNeuronsGrafted nigral neurons were found to have Lewy body-like inclusions14 years after transplantation into the striatum of an individual with PDOlanow CW.et al Nat Med.2008May;14(5):504-6.Transplanted dopamine neurons in people with PD do not contain Lewy bodiesMendez,Isacsonetal,NATUREMEDICINEVOLUME14(5):507-509,2008Freed CR,J Nucl Med.2010Jan;51(1):7-15-LongtermStudy-33oftheoriginaltrialparticipantswhowerefollowedfor2yearsaftertransplantationand15ofthesesubjectswhowerefollowedfor2additionalyears.-Theseresultssuggestthatclinicalbenefitandgraftviabilityaresustainedupto4yaftertransplantation.FreedCR,Neurotherapeutics(2011)8:549561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 CenterImprovedCellTherapyProtocolforParkinsonsDiseaseBasedonDifferentiationEfficiencyandSafetyofhESC-,HipscandNon-HumanPrimateiPSC-DerivedDANeuronsIsacsonetal,StemCells.2013;31(8):1548-62.Dopamine release from transplanted neural stem cells in Parkinsonian rat striatum in vivo.Zhouz,etal,ProcNatlAcadSciUSA.2014Nov4;111(44):15804-9iPSC-Derived Dopamine Neurons function after Transplantation in a Non-Human Primate Model of Parkinsons DiseaseCellStemCell.2015Mar5;16(3):269-74.OleIsacsonetal，HarvardStemCellInstituteStem cell-based Clinical Trials for(ALS)Nuralstem,Inc.thefirstPhaseIclinicaltrialforastemcell-basedtreatmentofALS.Initiatedin2010andcompletedin2013,involvedthetransplan-tationofhumanspinalcord-derivedNSCsintothespinalcordof15latetomid-stageALSpatientsGlass,Feldman,E.L.,2012.Lumbarintraspinalinjectionofneuralstemcellsinpatientswithamyotrophiclateralsclerosis:resultsofaphaseItrialin12patients.StemCells30(6),11441151.Riley,J.,Feldman,E.L.,2014.“IntraspinalstemcelltransplantationinALS:aphaseItrial,cervicalmicroinjectionandfinalsurgicalsafetyoutcomes”.Neurosurgery74(1),7787RESULTS:Unilateral cervical(group D,n=3)and cervical plus thoracolumbar(group E,n=3)microinjections to the ventral horn have been completed in ambulatory patients.One patient developed a postoperative kyphotic deformity prompting completion of a laminoplasty in subsequent patients.Another required reoperation for wound dehiscence and infection.The solitary patient with bulbar amyotrophic lateral sclerosis required perioperative reintubation.CONCLUSION:Delivery of a cellular payload to the cervical or thoracolumbar spinal cord was well tolerated by the spinal cord in this vulnerable population.This encouraging finding supports consideration of this delivery approach for neurodegenerative,oncologic,and traumatic spinal cord afflictions.IntraspinalstemcelltransplantationinALS:aphaseItrial,2014iPS Cells Were Generated from PD patients and Normal Controls 6-OHDA-induced Rat PD ModelHuman iPS cells Integrated to the Host Brain of 6-OHDA-induced Rat PD ModelHanF,WangW,ChenC,DuanJ,etalCytotherapy2015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。RT-PCR to Detect the MicroRNA Expression in Rat SCI Model MiR-124MiR-124MiR-124MiR-127MiR-127MiR-127MiR-127MiR-124MiR-133aMiR-133aMiR-133aMiR-181aMiR-181aMiR-181aReal-Time RT-PCR to Detect the MicroRNA Expression in SCI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):1264Cell.2012 September 14;150(6):126412731273BoneMarrowStromalCellIntraspinalTransplantsFailtoImproveMotorOutcomesinaSevereModelofSCIJournal of Neurotrauma 2015，Tuszynski MHlTodeterminewhetherlocalmechanismsmediateBMSCneuroprotectiveactionslgraftedallogeneicBMSCstositesofsevere,compressivespinalcordinjury(SCI)inSpragueDawleyrats.lCellswereadministered48hoursaftertheoriginalinjury.AdditionalanimalsreceivedallogeneicMSCsthatweregeneticallymodifiedtosecreteBDNF,tofurtherdeterminewhetheralocallyadministeredneurotrophicfactorprovidesorextendsneuroprotection.ltwomonthspost-injuryinaclinicallyrelevantmodelofsevereSCI,BMSCgraftswithorwithoutBDNFsecretionfailedtoimprovemotoroutcomes.Thus,allogeneicgraftsofBMSCsdonotappeartoactthroughlocalmechanisms,andfutureclinicaltrialsthatacutelydeliverBMSCstoactualsitesofinjurywithindaysareunlikelytobebeneficial.IntraspinalStemCellTransplantationinAmyotrophicLateralSclerosis:APhaseISafetyTrial,TechnicalNote,andLumbarSafetyOutcomesNEUROSURGERYVOLUME71|NUMBER2|AUGUST2012DepartmentofNeurosurgery,EmoryUniversity,Atlanta,Georgia;DepartmentofNeurology,EmoryUniversity,Atlanta,Georgia;DepartmentofNeurology,UniversityofMichigan,AnnArbor,Michigan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.LumbarLaminectomyMicroinjection platform applicationPostoperative imaging progressionRiley,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 87 Clinical Trials using ESCs and iPSCsThereisalsoareportofoneJapanesepatientwhoreceivedatransplantofasheetofiPSC-derivedRPESummary 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 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。致谢致谢/Acknowledgements北京大学生命科学研究中心康新江博士北京大学生命科学研究中心康新江博士干细胞与再生医学实验室全体科研人员：干细胞与再生医学实验室全体科研人员：王伟，张男，陈超，李森，卢现杰，段婧，吴士超，宋昊，王伟，张男，陈超，李森，卢现杰，段婧，吴士超，宋昊，宋娜，陈清法，刘延明宋娜，陈清法，刘延明干细胞与再生医学实验室研究团队干细胞与再生医学实验室研究团队Neural Differentiation of hUCB-MSC in VitrohUCB-MSC细胞移植细胞移植改善改善6-OHDA诱导诱导PD大鼠大鼠不对称运动行为不对称运动行为Differentiation of UC-MSC cells into Neurons and Dopamine Neurons