成髓鞘细胞研究和应用进展PPT课件

成髓鞘细胞研究和应用进展,安逸生活 pk 艰苦卓绝创业高原如此艰苦的环境：勇于担当，发挥引领作用以苦为乐，人生需要有一股精神,特朗普：我拒绝政治正确， 我只做正确的事情精英政治,2016.10.11. 中英脱髓鞘伦敦会议,要点,一些基本概念与影像学检查儿童脱髓鞘疾病临床分类成髓鞘细胞分类神经修复的路径各种细胞近期研究一览,Year 1977 2016 40年 demyelination remyelination Pubmed文献量趋势图,髓鞘构成成分,CNS 髓鞘中60 70% 为水分固体成分中脂类占70%, 蛋白质占30%构成髓鞘蛋白质主要两种成分：蛋白脂蛋白( PLP) 和髓鞘碱性蛋白(MBP)髓鞘相关糖蛋白(MAG) 髓鞘少突胶质细胞糖蛋白(MOG)Wolfram 蛋白脂DM 准大小蛋白其他微量蛋白,髓鞘功能,提供轴突与周围组织电绝缘，避免干扰相邻轴突之间轴突与其他结构之间加快动作电位传递：“跳跃式传导”机制引导受损轴突：再生,Myelin-related cells (Schwann cells and oligodendrocytes) co-operate with the axon in the formation and maintenance of myelin sheaths.,中枢神经系统(CNS)有髓神经纤维与周围(PNS)相似结构有髓鞘和郎飞结髓鞘外无基膜（神经膜）相邻神经纤维有时融合（箭头）,儿童发育,髓鞘化过程主要集中在：出生后18个月内（1.5年）1岁半接近成人oligodendrocyte少突胶质细胞：大多数细胞5-10年成熟，存活50年。每年减少1-300 OLS,The final stage of oligodendrocyte development is myelination. Unlike the peripheral nervous system where there is a strict size-dependent bias for myelination (only axons 1 um diameter enjoy the privilege), in the CNS axons as small as 300 nm are sometimes myelinated. Nevertheless, CNS myelination is also influenced by axonal size. Surprisingly, cultured oligodendrocytes will begin myelinating even synthetic axon-like tubes lacking the usual neuron-glia signaling cues. In this type of blind myelination, only tubes with a diameter of 400 nm were myelinated, suggesting this mode of myelination may be particularly important for larger axons.Moreover, once differentiating, it appears that oligodendrocytes only have a very narrow window of opportunity to select which adjacent axons to myelinate (5 h in the developing zebrafish and 12 h in myelinating cultures of rodent derived cells), irrespective of the total number of sheaths being made. This implies that the signaling pathways mediating this process are likely to be relatively robust. Although in vitro studies have implicated several key pathways, to date no one single molecule has been shown to be indispensable for myelination of CNS axons in vivo, highlighting the great degree of redundancy in the control of this vital process.Despite this, recent in vivo studies have revealed a great deal about how these various signaling pathways converge to control the extent or timing of myelination, if not overall myelination per se.,髓鞘脱失（脱髓鞘）一种very常见的临床神经病理变化,多指数T2/磁化转移MRI,provides an important step for understanding typical myelin development as well as providing the ability to identify when and where white matter abnormalities occur in neurodevelopmental disorders.myelin water volume fraction (MVF)髓磷脂体积分数,myelin water fraction (MWF)髓磷脂水分,脱髓鞘疾病分类（一）,儿童脱髓鞘病：周围性 和 中枢性周围性脱髓鞘病：多以双下肢或四肢瘫痪为首发中枢性脱髓鞘病：以视力障碍、肢体无力、发热、抽搐、头痛等为首发,脱髓鞘疾病分类（二）,经典病种：多发性硬化、急性播散性脑脊髓炎、慢性炎性脱髓鞘性多神经根神经病其他病种：缺血缺氧性脑病、脑性瘫痪、脊髓损伤、脑外伤、脑白质营养不良等神经创伤、变性、遗传和血管性疾病,Remyelination can occur in the damaged central nervous system (CNS),Since the discovery in the 1960s that remyelination can occur in the damaged CNS (Bunge et al. 1961) there has been much progress in understanding the cellular and molecular biology of oligodendroglia and the factors that regulate their propagation, migration, differentiation, maturation, and ability to myelinate nerve axons,脊髓损伤,脊髓挫伤1个月后，64的存活纤维发生脱髓鞘尽管有些轴突保持着解剖上的完整，但已无生理功能损伤部位远端的轴突出现华勒变性Franklin RJ, et al.1997,Spinal cord injury is accompanied by chronic progressive demyelination,Totoiu MO, Keirstead HS. 2005. Extent of demyelination and remyelination up to 450 days following contusive spinal cord injury in adult rats1 day post injury: the overall number of demyelinated axons peaked7-14 days post injury: declinedthen progressively increased up to 450 days post injuryOligodendrocyte and Schwann cell remyelinated axons appeared by 14 days post injuryremyelinated axons were present from 14 to 450 days post injury, remyelination was incomplete spinal cord injury is accompanied by chronic progressive demyelination,oligodendrocyte progenitor cell proliferationgeneration of new oligodendrocytesformation of thinner myelin,Domingues et al.2016.,Domingues et al.2016.,First research on transplantation of myelin-forming cells into the demyelinated spinal cord The cells survive, migrate, and find axons that need myelin insulation, and remyelinate them thereby restoring ability to conduct impulsesStephen Waxman and Jeffery Kocsis,细胞移植髓鞘化修复再生治疗脱髓鞘病理的神经修复重要策略和研究方向,嗅鞘细胞少突胶质细胞雪旺细胞多能诱导干（iPS）细胞神经干细胞胚胎干细胞脐带血/脐带间充质细胞骨髓间充质细胞脂肪干细胞皮肤干细胞,Olfactory ensheathing cell (OEC) 嗅鞘细胞,Remyelination,（1）嗅鞘细胞髓鞘化修复类于雪旺细胞,Pellitteri R, et al. (2010) OECs 能产生多种神经营养生长因子体外：能促进轴突生长体内：可形成髓鞘，促进髓鞘再生刺激轴突再生发芽优于雪旺细胞：与星形胶质细胞接触,（1）嗅鞘细胞髓鞘化修复类于雪旺细胞,Babiarz J, et al. (2011)分离幼年和成年大鼠的嗅球，分析表达GFP的OEC的髓鞘化轴突的能力。OEC：幼鼠OECs能髓鞘化背根神经节（DRG）轴突。嗅鞘细胞与轴突组装成束需要1周，如果形成可以检测到的轴突髓鞘，需要1周以上SC：大鼠雪旺细胞不捆束轴突，在1周内能形成P0 +和MBP +髓鞘节段大多数培养的OEC调宁蛋白（calponin）染色阳性，雪旺细胞为阴性几乎所有的OEC和雪旺细胞P75NTR和GFAP阳性两种细胞之间只有细微的免疫标记差异,（1）嗅鞘细胞髓鞘化修复类于雪旺细胞,Babiarz J, et al. (2011)The diameter of OEC generated myelin was greater than for Schwann cell myelin on DRG axons OEC but not Schwann cells myelinated DRG axons in the absence of vitamin C,（2）嗅鞘细胞修复脑缺血卒中,Shi X, et al. (2010)修复大脑中动脉闭塞（MCAO）大鼠脑白质 MCAO后56天结果：减少梗死体积，降低死亡率，改善神经功能缺损LFB髓鞘染色，NF免疫组织化学，Western blot：OEC移植大鼠髓鞘和轴突再生,（3）嗅鞘细胞修复周围神经损伤,Radtke C, et al. (2010) 嗅鞘细胞移植作为辅助治疗周围神经损伤嗅鞘细胞移植促进再生过程：介导趋化因子，神经营养和神经保护作用髓鞘形成：桥接，建立一个允许轴突再生的环境,（4）嗅鞘细胞在体内、体外与其他细胞的相互作用,Chuah MI, et al. (2010) 与其他类型的细胞：在体外和嗅鞘细胞移植后的胶质疤痕和炎症环境下的相互作用嗅鞘细胞和星形胶质细胞 克服胶质瘢痕的有害影响不同脊髓损伤的实验模型：OEC移植相关的胶质瘢痕的形态学改变在体外：嗅鞘细胞和胶质瘢痕的细胞类型组成之间的相互作用嗅鞘细胞：免疫细胞特性，移植到中枢神经系统损伤部位时，调制神经炎症,（5）嗅鞘细胞移植联合瘢痕切除修复脊髓挫伤,Zhang SX, et al. (2011)疤痕消融+ LP/ OEC移植促进大鼠脊髓慢性挫伤解剖恢复和P0（髓磷脂糖蛋白,P-zero）阳性髓鞘孟加拉玫瑰红光毒性方法 单独移植病灶腔：嗅黏膜固有层（LP）或联合体外培养的OECs,（6）嗅鞘细胞移植修复肌萎缩侧索硬化,Li Y, et al. (2011) OEC移植到脊髓：延长SOD1（G93A）ALS大鼠生存期神经保护作用和髓鞘化移植的嗅鞘细胞存活超过4周，在脊髓内迁移4.2毫米,（7）嗅鞘细胞移植修复各种脱髓鞘模型,Sasaki M, et al. (2011)不同的脱髓鞘环境对OEC髓鞘化修复的影响OECs的迁移和髓鞘形成：炎症处于活动状态炎症基本平息状态,（7）嗅鞘细胞移植修复各种脱髓鞘模型(2),Azimi Alamouti M, et al. Remyelinationof the corpus callosum byolfactoryensheathingcell in an experimental model of multiple sclerosis. (2015),（8）嗅鞘细胞移植修复效果的动物种属特异性,Wewetzer K, et al. (2011) 细胞增殖控制：种间差异鼠、狗、猪、猴、人Rodent： require mitogens for in vitro expansion a complex response to elevated intracellular cAMP, and undergo spontaneous immortalization upon prolonged mitogen stimulation,（9）胚胎/新生/成年嗅鞘细胞移植修复效果存在差异,Coutts DJ. Embryonic-derived olfactory ensheathing cells remyelinate focal areas of spinal cord demyelination more efficiently than neonatal or adult-derived cells. Cell Transplant. 2013 Form myelin sheathsOptimal donor age for OEC associated remyelination p75 purified OEC transplants from three donor agesolfactory bulbs of embryonic, neonatal, and adult rats and purified by immunopanningremyelinating potential was directly compared by transplantation into the same adult rat toxin-induced model of spinal cord demyelinationRemyelination efficiency 3 weeks after transplantation was assessed morphologically and by immunostainingall donor ages remyelinatethis process is most efficiently achieved by embryonic-derived OECs.,Oligodendrocyte少突胶质细胞,Oligodendrocyte precursor cells (OPCs), a subpopulation that accounts for 5 to 8% of cells within the central nervous system, are potential sources of oligodendrocyte replacement after SCI. OPCs react rapidly to injuries, proliferate at a high rate, and can differentiate into myelinating oligodendrocytes. However, posttraumatic endogenous remyelination is rarely complete.,Wang Y, et al. (2011),少突胶质祖细胞移植：在成年大鼠脊髓，趋向炎症区域存活分化成：可形成髓鞘的少突胶质细胞,Li H, et al. (2009),两层少突胶质细胞分化的转录调控 “双管齐下”的方式：创建一个基因控制的故障安全系统确保在发育过程、脱髓鞘病变的修复过程中：髓鞘化修复有序进行和有效明确地表达,Ishii A, et al. (2009),人类髓鞘蛋白质组学111 种已确认的蛋白质/转录物：在少突胶质细胞表达，在星形胶质细胞和神经元不表达163 additional proteinscomplexity of this metabolically active membrane,Sun F, et al. (2010),轴突变性对少突胶质细胞谱系细胞的影响： 背根切断术唤起：修复反应 脊髓挫伤后嘴侧轴突变性诱导：修复+细胞凋亡,Piaton G, et al. (2010),在发育髓鞘、脱髓鞘、修复过程中，轴突与少突胶质细胞相互作用髓磷脂的生物合成和髓鞘修复：神经元和少突胶质细胞之间的相互沟通必不可少In MS, CNS demyelination is often followed by spontaneous repair, mostly achieved by adult oligodendrocyte precursor cells. Extent of this myelin repair differs, ranging from very low, limited to the plaque border, to extensive, with remyelination throughout the shadow plaques. In addition to restoring neuronal connectivity, new myelin is neuroprotective. It reduces axonal loss and thus disability progression.,Neural stem cell (NSC) 神经干细胞,Hwang DH, et al. (2009),转导OLIG2转录因子的人类神经干细胞（NSCs）脊髓挫伤损伤后大鼠：提高运动功能的恢复增强脊髓白质髓鞘修复再生,Sher F, et al. (2009),生物发光成像（bioluminescence imaging）Olig2-NSCs增加在脱髓鞘小鼠模型的植入效果,Yang J, et al. (2010),比较骨髓和脑源性神经干细胞在中枢神经系统自身免疫性疾病的治疗效果类似a similar ability to differentiate into neurons, astrocytes, and oligodendrocytes both in vitro and in vivo both types of NSCs suppressed chronic experimental autoimmune encephalomyelitis therapeutic effects of NSCs include immunomodulation in the PNS and the CNS, neuron/oligodendrocyte repopulation by transplanted cells, and enhanced endogenous remyelination and axonal recovery,Carbajal KS, et al. (2010),多发性硬化病毒模型神经干细胞迁移通过CXCR4介导CXCL12的信号,Adipose mesenchymal stem cell脂肪来源间充质干细胞,Radtke C, et al. (2009),脂肪间充质干细胞衍生的神经球可分化成周围神经胶质样细胞,Constantin G, et al. (2009),脂肪来源的间质干细胞改善慢性实验性自身免疫性脑脊髓炎 (EAE)静脉给药：显著降低免疫反应所致EAE的严重性、减少脊髓炎和脱髓鞘、轴突损失ASC优先归巢到淋巴器官，迁移中枢神经系统双峰机制的治疗潜力：在疾病的早期阶段：抑制自身免疫反应诱导内源性祖细胞的神经再生,Age of the Donor Reduces the Ability of Human Adipose-Derived Stem Cells to Alleviate Symptoms in the Experimental Autoimmune Encephalomyelitis Mouse Model,ASCs from older donors failed to ameliorate the neurodegeneration associated with EAE, and mice treated with older donor cells had increased CNS inflammation, demyelination, splenocyte proliferation in vitro compared with the mice receiving cells from younger donors.,STEM CELLS TRANSLATIONALMEDICINE 2013;2:797,Ghasemi N. Transplantation of human adipose-derived stem cells enhances remyelination in lysolecithin-induced focal demyelination of rat spinal cord. Mol Biotechnol. 2014 .,transplanted human ADSCs (hADSCs) into a lysophosphatidylcholine (lysolecithin) model of multiple sclerosis (MS) and determined the efficiency of these cells in remyelination process. Forty adult rats were randomly divided into control, lysolecithin, vehicle, and transplantation groups, and focal demyelination was induced by lysolecithin injection into spinal cord. To assess motor performance, all rats were examined weekly with a standard EAE scoring scale. Four weeks after cell transplantation, to assess the extent of demyelination and remyelination, Luxol Fast Blue staining was used. In addition, immunohistochemistry technique was used for assessment of the presence of oligodendrocyte phenotype cells in damaged spinal cord. hADSCs had ability to differentiate into oligodendrocyte phenotype cells and improved remyelination process. Moreover, the evaluation of rat motor functions showed that animals which were treated with hADSC compared to other groups had significant improvement. hADSCs transplantation for cell-based therapies may play a proper cell source in the treatment of neurodegenerative diseases such as MS.,Co