1、神经退行性疾病干细胞移植治疗目前研究现状与未来展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望（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胎儿神经干细胞治疗帕金森氏病临床研究发展历程Evans JR,Mason SL,Barker RA.Prog Brain Res.2012;200:169-98 Lindvall O,et al，Nat Med.2008 May;14(5):501-3THSynucleinOverlayTransplanted fetal mesencephalic dopaminergic neurons(1
2、1-16 years)developed alpha-synuclein-positive Lewy bodies in grafted neuronsSynuclein-HostUbiquintin-HostSynuclein-Grafted NeuronsUbiquintin-Grafted NeuronsGrafted nigral Grafted nigral neurons were found to have Lewy were found to have Lewy body-like inclusions14 years after transplantation body-li
3、ke inclusions14 years after transplantation into the striatum of an individual with 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 Transplanted dopamine neurons in people with PD do not contain Lewy bodiesdo not c
4、ontain Lewy bodiesMendez,Isacson et al,NATURE MEDICINE VOLUME 14(5):507-509,2008Freed CR,Freed CR,J Nucl Med.2010 Jan;51(1):7-15-Long term Study-33 of the original trial participants who were followed for 2 years after transplantation and 15 of these subjects who were followed for 2 additional years
5、.-These results suggest that clinical benefit and graft viability are sustained up to 4 y after transplantation.Freed CR,Neurotherapeutics(2011)8:549 561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 Slo
6、an-Kettering Cancer CenterImproved Cell Therapy Protocol for Parkinsons Disease Based on Differentiation Efficiency and Safety of hESC-,Hipsc and Non-Human Primate iPSC-Derived DA NeuronsIsacson et al,Stem Cells.2013;31(8):1548-62.Dopamine release from transplanted neural stem cells in Parkinsonian
7、rat striatum in vivo.Twelve patients were treated with either unilateral or bilateral injections.6OHDAinduced Rat PD Model分化的胎脑神经干细胞移植治疗PDIncreasing host neurogenesis“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.To determine
8、 whether local mechanisms mediate BMSC neuroprotective actionset al Nat Med.（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Release of immuno-regulatory factors such as IL-2,6,8,10 to play immuno-modulation and attenuation of the inflammatory process,such as MSC.2012;200:169-98Improved Cell Thera
9、py Protocol for Parkinsons Disease Based on Differentiation Efficiency and Safety of hESC-,Hipsc and Non-Human Primate iPSC-Derived DA NeuronsIntraspinal stem cell transplantation in ALS:a phase I trial,2014Another required reoperation for wound dehiscence and infection.using ESCs and iPSCsiPS Cells
10、 Were Generated from PD patients and Normal ControlsUbiquintin-Grafted NeuronsIncreasing host neurogenesisgrafted allogeneic BMSCs to sites of severe,compressive spinal cord injury(SCI)in Sprague Dawley rats.Dopamine release from transplanted neural stem Dopamine release from transplanted neural ste
11、m cells in Parkinsonian rat striatum in vivo.cells in Parkinsonian rat striatum in vivo.Zhou z,et al,Proc Natl Acad Sci U S A.2014 Nov 4;111(44):15804-9iPSCDerived Dopamine Neurons function iPSCDerived Dopamine Neurons function after Transplantation in a NonHuman after Transplantation in a NonHuman
12、Primate Model of Parkinsons DiseasePrimate Model of Parkinsons Disease Cell Stem Cell.2015 Mar 5;16(3):269-74.Ole Isacson et al，Harvard Stem Cell InstituteStem cellbased Clinical Trials for(ALS)Nuralstem,In c.the first Phase I clinical trial for a stem cell-based treatment of ALS.Initiated in 2010 a
13、nd completed in 2013,involved the transplan-tation of human spinal cord-derived NSCs into the spinal cord of 15 late to mid-stage ALS patients“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.Summary on Molecular Mechanism of St
14、em Cell Transplantation for Neurological Diseasesto increase the functions of the endogenous neural stem cellsThis encouraging finding supports consideration of this delivery approach for neurodegenerative,oncologic,and traumatic spinal cord afflictions.暴露和部分横切脊髓外科手术。University,Atlanta,Georgia;Depar
15、tment of Neurology,Emory University,Atlanta,Georgia;Department of Neurology,University of Michigan,Ann Arbor,MichiganGlass,Feldman,E.RTPCR to Detect the MicroRNA Expression in Rat SCI ModelPatients are followed clinically and radiologically to assess potential toxicity of the procedure.Cells were ad
16、ministered 48 hours after the original injury.SC grafts should exhibit regulated release of dopamine in line with that of endogenous dopaminergic neurons.-Long term Study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Isacson et al,Stem Cells.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RealTime RTPCR to Detect the MicroRNA Expression in SCI
17、iPS Cells Were Generated from PD patients and Normal ControlsGlass,Feldman,E.Thus,allogeneic grafts of BMSCs do not appear to act through local mechanisms,and future clinical trials that acutely deliver BMSCs to actual sites of injury within days are unlikely to be beneficial.Ubiquintin-Grafted Neur
18、ons“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.This encouraging finding supports consideration of this delivery approach for neurodegenerative,oncologic,and traumatic spinal cord afflictions.Intraspinal stem cell transplan
19、tation in ALS:a phase I trial,2014 Glass,Feldman,E.L.,2012.Lumbar intraspinal injection of neural stem cells in patients with amyotrophic lateral sclerosis:results of a phase I trial in 12 patients.Stem Cells 30(6),1144 1151.Riley,J.,Feldman,E.L.,2014.“Intraspinal stem cell transplantation in ALS:a
20、phase I trial,cervical microinjection and final surgical safety outcomes”.Neurosurgery 74(1),77 87RESULTS: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 k
21、yphotic 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
22、 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.Intraspinal stem cell transplantation in ALS:a phase I tri
23、al,2014iPS Cells Were Generated from PD patients iPS Cells Were Generated from PD patients and Normal Controls and Normal Controls 6OHDAinduced Rat PD ModelHuman iPS cells Integrated to the Host Human iPS cells Integrated to the Host Brain of 6OHDAinduced Rat PD ModelBrain of 6OHDAinduced Rat PD Mod
24、elHan F,Wang W,Chen C,Duan J,et al Cytotherapy 2015 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PDPD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。RTPCR to Detect the MicroRNA Expression in Rat RTPCR to Detect the MicroRNA Expression in Rat SCI Model
25、SCI Model MiR-124MiR-124MiR-124MiR-127MiR-127MiR-127MiR-127MiR-124MiR-133aMiR-133aMiR-133aMiR-181aMiR-181aMiR-181aRealTime RTPCR to Detect the RealTime RTPCR to Detect the MicroRNA Expression in SCIMicroRNA 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
26、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):12641273Bone Marrow Stromal Cell Intraspinal Transplants Fail to Improve Motor Outcomes in a Severe Model of SCIJournal of Neurotrauma 2015，Tuszynski MHl To determine whether local mechanisms m
27、ediate BMSC neuroprotective actions l grafted allogeneic BMSCs to sites of severe,compressive spinal cord injury(SCI)in Sprague Dawley rats.l Cells were administered 48 hours after the original injury.Additional animals received allogeneic MSCs that were genetically modified to secrete BDNF,to furth
28、er determine whether a locally administered neurotrophic factor provides or extends neuroprotection.l two months post-injury in a clinically relevant model of severe SCI,BMSC grafts with or without BDNF secretion failed to improve motor outcomes.Thus,allogeneic grafts of BMSCs do not appear to act t
29、hrough local mechanisms,and future clinical trials that acutely deliver BMSCs to actual sites of injury within days are unlikely to be beneficial.Intraspinal Stem Cell Transplantation in Amyotrophic Lateral Sclerosis:A Phase I SafetyTrial,Technical Note,and Lumbar Safety OutcomesNEUROSURGERY VOLUME
30、71|NUMBER 2|AUGUST 2012Department of Neurosurgery,EmoryUniversity,Atlanta,Georgia;Department of Neurology,Emory University,Atlanta,Georgia;Department of Neurology,University of Michigan,Ann Arbor,Michigan神经干细胞移植方法神经干细胞移植方法 Each microinjection series comprised 5 injections(10mL/injection)separated by
31、 4 mm.Each injection:100 000 neural stem cells derived from a fetal spinal cord.Twelve patients were treated with either unilateral or bilateral injections.Patients are followed clinically and radiologically to assess potential toxicity of the procedure.Lumbar LaminectomyMicroinjection platform appl
32、icationMicroinjection 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 Clinical Trials using ESCs and iPSC
33、susing ESCs and iPSCsThere is also a report of one Japanese patient who received a transplant of asheet of iPSC-derived RPE Summary on Molecular Mechanism of Stem Cell Transplantation for Neurological Diseases Transplanted cells survive，differentiate to neurons,astrocytes,oligodendrocyte precursors(
34、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 cells Release of immuno-regulatory factors such as IL-2,6,8,10 to play immuno-modulation and attenuation of the inflammator
35、y 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 Ole Isacson et al，Harvard Stem Cell InstituteEach injection:100 000 neural stem cells derived from a fetal spinal cord.Cells
36、 were administered 48 hours after the original injury.sheet of iPSC-derived RPEOthers such as delaying the onset and prolonging survival of SOD1 ratsEach microinjection series comprised 5 injections(10mL/injection)separated by 4 mm.“Intraspinal stem cell transplantation in ALS:a phase I trial,cervic
37、al microinjection and final surgical safety outcomes”.Summary on Molecular Mechanism of Stem Cell Transplantation for Neurological DiseasesIntraspinal stem cell transplantation in ALS:a phase I trial,2014Glass,Feldman,E.University,Atlanta,Georgia;Department of Neurology,Emory University,Atlanta,Geor
38、gia;Department of Neurology,University of Michigan,Ann Arbor,MichiganAmyotrophic Lateral Sclerosis:Cells were administered 48 hours after the original injury.There is also a report of one Japanese patient who received a transplant of a,Feldman,E.Amyotrophic Lateral Sclerosis:2008May;14(5):504-6.“Int
39、raspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.Twelve patients were treated with either unilateral or bilateral injections.Mendez,Isacson et al,NATURE MEDICINE VOLUME 14(5):507-509,2008干细胞移植修复脊髓神经损伤Amyotrophic Lateral Sclerosis:O
40、le Isacson et al，Harvard Stem Cell InstituteIntraspinal stem cell transplantation in ALS:a phase I trial,2014“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.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Patients are followed clinically and radiolo
41、gically to assess potential toxicity of the procedure.Ole Isacson et al，Harvard Stem Cell InstituteThis include s the absence of tumor formation and GIDs（graf t-induced dyskinesia）throughout long-term follow-up periods.2012 September 14;150(6):12641273Ubiquintin-Grafted NeuronsIncreasing host neurog
42、enesisEvaluation System（Symptoms,Dopamine release,Levodopa-response),Unified Parkinson s Disea se Ra ting Scal e(UPDRS)，BiomarkersTrial,Technical Note,and Lumbar Safety Outcomes“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.,
43、Feldman,E.Adverse effects must be minimal.Grafted nigral neurons were found to have Lewy body-like inclusions14 years after transplantation into the striatum of an individual with PD“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcom
44、es”.“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.Dopamine release from transplanted neural stem cells in Parkinsonian rat striatum in vivo.分化的胎脑神经干细胞移植治疗PDiPS Cells Were Generated from PD patients and Normal Controlsto incr
45、ease the functions of the endogenous neural stem cells6OHDAinduced Rat PD Model2010 Jan;51(1):7-15“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.There is also a report of one Japanese patient who received a transplant of a-Lo
46、ng term StudyThere is also a report of one Japanese patient who received a transplant of aPatients are followed clinically and radiologically to assess potential toxicity of the procedure.Patients are followed clinically and radiologically to assess potential toxicity of the procedure.SC grafts shou
47、ld exhibit regulated release of dopamine in line with that of endogenous dopaminergic neurons.A Phase I Safety暴露和部分横切脊髓外科手术。“Intraspinal stem cell transplantation in ALS:a phase I trial,cervical microinjection and final surgical safety outcomes”.Zhou z,et al,Proc Natl Acad Sci U S A.Intraspinal Stem
48、 Cell Transplantation inCell Sources：Neural projenitors，MSC，hES cells，iPS cellsSC grafts should exhibit regulated release of dopamine in line with that of endogenous dopaminergic neurons.Lorenz Studer，et al Memorial Sloan-Kettering Cancer Center2012;200:169-98-Long term StudyCells were administered
49、48 hours after the original injury.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 o
50、f 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 （
