Lentiviral vector mediated Nurrl gene-modified mesenchymal stem cells for the treatment of Parkinson’s disease rat model

doi:10.16098/j.issn.0529-1356.2015.06.004

Abstract

Abstract:

Objective To investigate the therapeutic effects of bone marrow mesenchymal stem cells (BMSCs) modified by nuclear receptor-related factor 1 (Nurrl) gene transplanted into the striatum of Parkinson’s disease rat model. Methods Eighteen rat models of Parkinson’s disease (PD) were successfully induced by 6-hydroxydopamine (6-OHDA) injection into the right striatum. BMSCs infected by GV287-Nurr1 lentivirus carrying green fluorescence were transplanted into the striatum of six PD rats (the experiment group). Randomly set up six PD rats as saline group (the sham-grafted group) and six PD rats as negative lentivirus GV287 infection of BMSCs group (the control group). After 1, 2 and 4 weeks, the behavioral change of the rats was induced by apomorphine (APO). The expression change of Nurrl and TH in the striatum and substantia nigra were examined by immunohistochemical staining, and the expression change of Nurrl mRNA and TH mRNA in the striatum and substantia nigra were detected by RT-PCR. Results The Nurr1 protein was detected in BMSCs and its supernatant after infected by lentivirus GV287-Nurr1. After transplantation 1, 2 and 4 weeks compared to the sham-grafted group the rat rotational behavior of the control group and the experiment group were improved, and that of the experiment group improved more obvious. The implanted Nurr1-positive cells survived and migrated to the cortex and the contralateral brain tissue along the corpus callosum. Compared with the sham-grafted group and the control group, the expression of Nurrl and TH at protein and mRNA levels of the lesion side of the striatum and substantia nigra after transplantation in experiment group were increased significantly. Conclusion Lentiviral vector mediated Nurrl gene-modified BMSCs can effectively improve the symptoms of PD rats and increase the expression of Nurrl and TH in striatum and substantia nigra of Parkinson’s disease rat model.

Key words: Bone marrow mesenchymal stem cell, Lentivirus, Nuclear receptor-related factor 1, Tyrosine hydroxylase, Parkinson’s disease, Immunohistochemistry, Rat

WANG Xiao-xiao FU Wen-yu* ZHUANG Wen-xin LI Feng-jie WANG Qian LIU Jin-cheng. Lentiviral vector mediated Nurrl gene-modified mesenchymal stem cells for the treatment of Parkinson’s disease rat model[J]. AAS, 2015, 46(6): 742-749.

References

［1］Politis M, Lindvall O. Clinical application of stem cell therapy in Parkinson’s disease［J］. BMC Med， 2012, 10: 1.
［2］Fu W, Zheng Z, Zhuang W, et al. Neural metabolite changes in corpus striatum after rat multipotent mesenchymal stem cells transplanted in hemiparkinsonian rats by magnetic resonance spectroscopy［J］. Int J Neurosci, 2013, 123(12): 883-891.
［3］Krampera M, Pasini A, Pizzolo G, et al. Regenerative and immunomodulatory potential of mesenchymal stem cells［J］. Curr Opin Pharmacol, 2006, 6(4):435-441.
［4］Wang Z, Benoit G, Liu J, et al. Structure and function of Nurr1 identifies a class of ligand-independent nuclear receptors［J］. Nature, 2003, 423(6939): 555-560.
［5］Brito-Armas JM, Baekelandt V, Castro-Hernández JR, et al. Melatonin prevents dopaminergic cell loss induced by lentiviral vectors expressing A30P mutant alpha-synuclein［J］. Histol Histopathol, 2013, 28(8): 999-1006. 
［6］Chen SS, Yang C, Hao F, et al. Intrastriatal GDNF gene transfer by inducible lentivirus vectors protects dopaminergic neurons in a ratmodel of parkinsonism［J］. Exp Neurol, 2014 261:87-96.
［7］Palfi S, Gurruchaga JM, Ralph GS, et al. Long-term safety and tolerability of ProSavin, a lentiviral vector-based gene therapy for Parkinson’s disease: a dose escalation, open-label, phase 1/2 trial［J］. Lancet, 2014, 383(9923): 1138-1146. 
［8］Paxinos G, Watson C. The Rat Brain Stereotaxic Coordinates［M］. Sydney:Academic Press Sydney, 1985: 87-92.
［9］Wang XX, Fu WY, Zhuang WX, et al. Construction and identification of lentiviral vector over-expressing Nr4a2［J］. Biotech World, 2014, 11(10): 84-85.（in Chinese）
王晓晓, 付文玉, 庄文欣, 等. Nr4a2 过表达慢病毒载体的构建及鉴定［J］. 生物技术世界, 2014, 11(10): 84-85.
［10］Wang XC, Wu JSh, Fu WY, et al. Morphologic characteristics of neuron-like cells induced from the rat bone marrow mesenchymal stem cells［J］. Acta Anatomica Sinica, 2007, 38(1): 107-110.（in Chinese）
王晓萃, 吴金生, 付文玉, 等. 诱导大鼠间充质干细胞形成的神经元样细胞的形态特征［J］. 解剖学报, 2007, 38(1): 107-110.
［11］Luan ShJ, Fu WY, Zhuang WX, et al. Expression changes of 5-HT, NPY, SP and its mRNA in dorsal raphe nucleus of rats with Parkinson’s disease［J］. Chinese Journal of Anatomy, 2014, 37(1): 58-62.（in Chinese）
栾守婧, 付文玉, 庄文欣, 等. 帕金森病大鼠中缝背核5-羟色胺、神经肽Y、P物质蛋白及mRNA的表达变化［J］. 解剖学杂志, 2014, 37(1): 58-62.
［12］Nakata Y, Yasuda T, Mochizuki H. Recent progress in gene therapy for Parkinson’s disease［J］. Curr Mol Med, 2012, 12(10): 1311-1318. 
［13］Verstraeten A, Theuns J, Van Broeckhoven C. Progress in unraveling the genetic etiology of Parkinson disease in a genomic era［J］. Trends Genet, 2015, 31(3): 140-149. 
［14］Park HJ, Shin JY, Lee BR, et al. Mesenchymal stem cells augment neurogenesis in the subventricular zone and enhance differentiation of neural precursor cells into dopaminergic neurons in the substantia nigra of a parkinsonian model［J］. Cell Transplant, 2012, 21(8): 1629-1640.
［15］Venkataramana NK, Pal R, Rao SA, et al. Bilateral transplantation of allogenic adult human bone marrow-derived mesenchymal stem cells into the subventricular zone of parkinson’s disease: a pilot clinical study［J］. Stem Cells Int, 2012, 2012: 931902.
［16］Grinev VV, Severin IN, Posrednik DV, et al. Highly efficient transfer and stable expression of two genes upon lentivirus transduction of mesenchymal stem cells from human bone marrow［J］. Genetika, 2012, 48(3): 389-400.
［17］Xu SH, Song ChJ, Pan Y. Gene transfer into mesenchymal stem cells:a comparison between the adenoviral and lentiviral vector delivery systems［J］. Life Science Research, 2013, 17(6): 543-547.（in Chinese）
徐思慧, 宋春敬, 潘晔. 腺病毒载体与慢病毒载体感染骨髓间充质干细胞的比较［J］. 生命科学研究, 2013, 17(6): 543-547.
［18］Heng X,Jin G, Zhang X,et al. Nurr1 regulates Top IIβand functions in axon genesis of mesencephalic dopaminergic neurons［J］. Mol Neurodegener, 2012, 7: 4.
［19］Kadkhodaei B, Alvarsson A, Schintu N, et al. Transcription factor Nurr1 maintains fiber integrity and nuclear-encoded mitochondrial gene expression in dopamine neurons［J］. Proc Natl Acad Sci USA, 2013, 110(6): 2360-2365.
［20］Alvarez-Castelao B, Losada F, Ahicart P, et al. The N-terminal region of Nurr1 (a.a 1-31) is essential for its efficient degradation by the ubiquitin proteasome pathway［J］. PLoS One, 2013, 8(2): e55999. 
［21］García-Yagüe ?J, Rada P, Rojo AI, et al. Nuclear import and export signals control the subcellular localization of Nurr1 protein in response to oxidative stress［J］. J Biol Chem, 2013, 288(8): 5506-5517. 
［22］Xu HW, Sun ShL. Comparison of transplantation of mesenchymal stem cells and Nurrl-overexpressing mesenchym al stem cells with fetal ventral mesencephalic cells in Parkinson disease rat model［J］. Journal of Chinese Practical Diagnosis and Therapy, 2011, 25(10): 967-969.（in Chinese）
徐浩文, 孙石磊. 骨髓间充质干细胞、过表达Nurr1 蛋白的骨髓间充质干细胞与胎脑细胞移植治疗帕金森病大鼠的比较［J］. 中华实用诊断与治疗杂志, 2011, 25(10): 967-969.

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