Msx3基因特异诱导小胶质细胞选择性极化的方法及其应用
【技术领域】
[0001] 本发明属于基因工程和医学领域，具体涉及一种高表达同源盒基因 MSX3 (msh-like homeobox-3)的小胶质细胞及其在多发性硬化等脱髓鞘疾病中促进再髓鞘 化的应用。
【背景技术】
[0002] 多发性硬化（MS)是一种自身免疫性中枢神经系统脱髓鞘疾病。大部分患者以 复发缓解型起病，之后发展成不可逆的继发进展型（Trapp, B.D., and Nave, K.A. 2008. Multiple sclerosis: an immune or neurodegenerative disorder ? Annu Rev Neurosci 31:247-269.)。机体修复性的再髓鞘只出现在MS的早期阶段，目前认为晚期再髓鞘功 能的丧失很可能是导致MS由复发缓解型发展成不可逆的继发进展型的重要原因 （Scol ding, N. , Franklin, R. , Stevens, S. , Heldinj C. Η. , Compstonj A. , and Newcombej J. 1998. Oligodendrocyte progenitors are present in the normal adult human CNS and in the lesions of multiple sclerosis. Brain 121 (Pt 12) :2221-2228.) D 而再髓鞘的失败很可 能是由MS慢性病灶处的神经轴突退行性变（Zamvil，S. S.，and Steinman，L 2003. Diverse targets for intervention during inflammatory and neurodegenerative phases of multiple sclerosis. Neuron 38:685-688.)和少突胶质细胞前体细胞分化障碍（Kuhlmann ,T. , Miron, V. , Cuij Q. , Wegner, C. , Antelj J. , and Bruckj W. 2008. Differentiation block of oligodendroglial progenitor cells as a cause for remyelination failure in chronic multiple sclerosis. Brain 131:1749-1758. ；Shen, S. , Sandoval, J. , Swiss, V. A. , Li, J. , Dupree, J. , Franklin, R. J. , and Casaccia-Bonnefilj P. 2008. Age-dependent epigenetic control of differentiation inhibitors is critical for remyelination efficiency. Nat Neurosci 11:1024-1034·)导致的〇
[0003] 目前发现小胶质细胞的极化在调节神经元和少突胶质细胞前体细胞功能中发挥 重要作用（Peferoen，L.，Kipp，M.，van der Valk，P.，van Noort，J.M.，and Amor，S.2013. Review series on immune responses in neurodegenerative diseases:Oligodendro cyte-microglia cross-talk in the CNS. Immunology. ；Minghetti,L. 2005. Role of inflammation in neurodegenerative diseases. Curr Opin Neuroll8:315-321. ；Rasm ussen, S. , Wang, Y. , Kivisakkj P. , Bronson, R. T. , Meyer, M. , Imitolaj J. , and Khouryj S. J. 2007.Persistent activation of microglia is associated with neuronal dysfunction of callosal projecting pathways and multiple sclerosis-like lesions in relapsing-remitting experimental autoimmune encephalomyelitis. Brain 130:2816-2829.)。不同极化状态的小胶质细胞分泌不同细胞因子促进少 突胶质细胞前体细胞分化，并保护神经元免予损伤（Liao, B.，Zhao, W.，Beers, D. R. , Henkel,J.S., and Appel,S. H. 2012. Transformation from a neuroprotective to a neurotoxic microglial phenotype in a mouse model of ALS. Exp Neurol 237:147-152. ;Miron，V. E.，Boyd，A.，Zhao, J. W.，Yuen，T. J.，Ruckh，J. M.，Shadrach，J. L. , van Wijngaardenj P. , Wagers, A. J. , Williams, A. , Franklin, R. J. , et al. 2013. M2microglia and macrophages drive oligodendrocyte differentiation during CNS remyelination. Nat Neurosci 16:1211-1218.；
[0004] Butovsky, 0. , Landaj G. , Kunisj G. , Zivj Y. , Avidanj H. , Greenberg, N. , Schwa rtz, A. , Smirnov, I. , Pollack, A. , Jung, S. , et al. 2006. Induction and blockage of oligodendrogenesis by differently activated microglia in an animal model of multiple sclerosis. J Clin Invest 116:905-915. ；Kigerl, K. A. ,Genselj J. C. , Ankeny, D. P. , Alexander, J. K. , Donnelly, D. J. , and Popovich, P. G. 2009. Identification of two distinct macrophage subsets with divergent effects causing either neurotoxicity or regeneration in the injured mouse spinal cord. J NeuiOsci 29:13435-13444.)。Ml型小胶质细胞数量同轴突损伤程度正相关，而M2型 只在再髓鞘病灶及病灶周围存在（1^81111188611，3.，？3叫，￥.，1(；!_￥1831^，？.，81'0118011，1?· T. , Meyer, M. , Imitolaj J. , and Khouryj S. J. 2007. Persistent activation of microglia is associated with neuronal dysfunction of callosal projecting pathways and multiple sclerosis-like lesions in relapsing-remitting experimental autoimmune encephalomyelitis. Brainl30: 2816-2829. ；Miron, V. E. , Boyd, A. , Zhao, J. W. , Yuen, T. J. , Ruckhj J. M. , Shadrachj J. L. , van Wijngaardenj P. , Wagers, A. J. , Williams,A., Franklin,R.J. ,et al.2013.M2microglia and macrophages drive oligodendrocyte differentiation during CNS remyelination.Nat Neurosci 16:1211-1218. ;Bitsch，A.，Schuchardt，J.，Bunkowski，S.，Kuhlmann，T. ，and Bruckj W. 2000. Acute axonal injury in multiple sclerosis. Correlation with demyelination and inflammation. Brain 123 (Pt6): 1174-1183.)〇 由此， 阻断Ml向M2的极化很可能是导致慢性非活动的MS病灶处再髓鞘失败的重要 因素（Miron，V. E.，Boyd，A.，Zhao, J. W.，Yuen，T. J.，Ruckh，J. M.，Shadrach，J. L. , van Wijngaardenj P. , Wagers, A. J. , Williams,