Msx3基因特異誘導(dǎo)小膠質(zhì)細(xì)胞選擇性極化的方法及其應(yīng)用
【技術(shù)領(lǐng)域】
[0001] 本發(fā)明屬于基因工程和醫(yī)學(xué)領(lǐng)域，具體涉及一種高表達(dá)同源盒基因 MSX3 (msh-like homeobox-3)的小膠質(zhì)細(xì)胞及其在多發(fā)性硬化等脫髓鞘疾病中促進(jìn)再髓鞘 化的應(yīng)用。
【背景技術(shù)】
[0002] 多發(fā)性硬化（MS)是一種自身免疫性中樞神經(jīng)系統(tǒng)脫髓鞘疾病。大部分患者以 復(fù)發(fā)緩解型起病，之后發(fā)展成不可逆的繼發(fā)進(jìn)展型（Trapp, B.D., and Nave, K.A. 2008. Multiple sclerosis: an immune or neurodegenerative disorder ? Annu Rev Neurosci 31:247-269.)。機(jī)體修復(fù)性的再髓鞘只出現(xiàn)在MS的早期階段，目前認(rèn)為晚期再髓鞘功 能的喪失很可能是導(dǎo)致MS由復(fù)發(fā)緩解型發(fā)展成不可逆的繼發(fā)進(jìn)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 而再髓鞘的失敗很可 能是由MS慢性病灶處的神經(jīng)軸突退行性變（Zamvil，S. S.，and Steinman，L 2003. Diverse targets for intervention during inflammatory and neurodegenerative phases of multiple sclerosis. Neuron 38:685-688.)和少突膠質(zhì)細(xì)胞前體細(xì)胞分化障礙（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·)導(dǎo)致的〇
[0003] 目前發(fā)現(xiàn)小膠質(zhì)細(xì)胞的極化在調(diào)節(jié)神經(jīng)元和少突膠質(zhì)細(xì)胞前體細(xì)胞功能中發(fā)揮 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.)。不同極化狀態(tài)的小膠質(zhì)細(xì)胞分泌不同細(xì)胞因子促進(jìn)少 突膠質(zhì)細(xì)胞前體細(xì)胞分化，并保護(hù)神經(jīng)元免予損傷（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型小膠質(zhì)細(xì)胞數(shù)量同軸突損傷程度正相關(guān)，而M2型 只在再髓鞘病灶及病灶周圍存在（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， 阻斷Ml向M2的極化很可能是導(dǎo)致慢性非活動(dòng)的MS病灶處再髓鞘失敗的重要 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,