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 (11-16 years) developed alpha-synuclein-posit
2、ive Lewy bodies in grafted neuronsSynuclein-HostUbiquintin-HostSynuclein-Grafted NeuronsUbiquintin-Grafted NeuronsGrafted Grafted nigralnigral neurons were found to have were found to have LewyLewy body-like inclusions14 years after transplantation body-like inclusions14 years after transplantation
3、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 contain Lewy bodiesMendez, Isacson et al, ,
4、 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. - These results suggest that
5、 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 Sloan-Kettering Cance
6、r 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 Dopamine release from transplanted
7、 neural stem 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):15804-9iPSC-Derived Dopamine Neurons iPSC-Derived Dopamine Neurons function after Transplantation in a Non-function after Transplantation in
8、 a Non-Human Primate Model of Parkinsons Human Primate Model of Parkinsons DiseaseDisease Cell Stem Cell. 2015 Mar 5;16(3):269-74. Ole Isacson et al，Harvard Stem Cell InstituteStem cell-based Clinical Trials for (ALS) Nuralstem, In c . the first Phase I clinical trial for a stem cell-based treatment
9、 of ALS. Initiated in 2010 and 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 Glass, Feldman, E.L., 2012. Lumbar intraspinal injection of neural stem cells in patients with amyotrophic lateral sclerosis: re
10、sults 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 phase I trial, cervical microinjection and final surgical safety outcomes”. Neurosurgery 74 (1), 77 87RESULTS: Unilateral cervical (group D, n = 3)
11、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 a
12、nd 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 suppo
13、rts consideration of this delivery approach for neurodegenerative, oncologic, and traumatic spinal cord afflictions. Intraspinal stem cell transplantation in ALS: a phase I trial, 2014IPS CELLS WERE GENERATED FROM PD IPS CELLS WERE GENERATED FROM PD PATIENTS AND NORMAL CONTROLS PATIENTS AND NORMAL C
14、ONTROLS 6-OHDA-induced Rat PD ModelHUMAN IPS CELLS INTEGRATED TO THE HUMAN IPS CELLS INTEGRATED TO THE HOST BRAIN OF 6-OHDA-INDUCED RAT HOST BRAIN OF 6-OHDA-INDUCED RAT PD MODELPD MODELHan 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
15、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 RT-PCR to Detect the MicroRNA Expression in Rat SCI Model Rat SCI Model MiR-124MiR-124MiR-124MiR-127MiR-127MiR-127MiR-127MiR-124MiR-133aMiR-133aMiR-133aMiR-181aMiR-1
16、81aMiR-181aReal-Time RT-PCR to Detect the Real-Time RT-PCR to Detect the MicroRNA Expression in SCIMicroRNA Expression in SCI051015202530MiR-1MiR-17MiR-21MiR-30d系列系列1d0d7d1d1400.20.40.60.811.21.41.61.82MiR-124MiR-127MiR-133aMiR-181ad0d7d1dd7d1d14d0d7d1dd7d1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
17、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): 12641273Bone Marrow Stromal Cell Intraspinal Transplants Fail to Improve Motor Outcomes in a Severe Model of SCIJournal of Neurotrauma 2015， Tuszynski MHl
18、 To determine whether local mechanisms mediate 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
19、genetically modified to secrete BDNF, to further 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,
20、 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. Intraspinal Stem Cell Transplantation in Amyotrophic Lateral Sclerosis: A Phase I SafetyTrial, Technic
21、al Note, and Lumbar Safety OutcomesNEUROSURGERY VOLUME 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
22、 microinjection series comprised 5 injections (10mL/injection) separated by 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 pot
23、ential toxicity of the procedure.Lumbar LaminectomyMicroinjection platform applicationMicroinjection 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
24、outcomes”. Neurosurgery 74 (1), 77 87 Clinical Trials Clinical Trials using ESCs and iPSCsusing 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 T
25、ransplanted 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 cells Release of immuno-
26、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
27、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 dopam
28、inergic 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 （Sympto
29、ms, 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/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
