Hoban D B, Howard L, Dowd E
Pharmacology & Therapeutics, National University of Ireland, Galway, Ireland; NCBES Galway Neuroscience Centre, National University of Ireland, Galway, Ireland.
The Regenerative Medicine Institute, National University of Ireland, Galway, Ireland.
Neuroscience. 2015 Sep 10;303:402-11. doi: 10.1016/j.neuroscience.2015.07.014. Epub 2015 Jul 10.
Constraints involving the delivery method of glial cell line-derived neurotrophic factor (GDNF) have hampered its efficacy as a neuroprotectant in Parkinson's disease. Ex vivo gene therapy, in which suitable cells, such as bone marrow-derived mesenchymal stem cells (MSCs), are genetically engineered to overexpress GDNF (GDNF-MSCs) prior to transplantation may be more beneficial than direct brain infusion of the neurotrophin. Previously, GDNF-MSCs have been assessed in the commonly employed 6-hydroxydopamine neurotoxic model of Parkinson's disease. In this study however, we used an emerging inflammatory model of Parkinson's disease (the lipopolysaccharide (LPS) model) to assess the ability of transplanted GDNF-MSCs to protect against LPS-induced neuroinflammation, neurodegeneration and behavioral impairment. Thirty male Sprague-Dawley rats were used in this experiment. Rats were performance matched based on baseline motor function tests into three groups (LPS lesion only, LPS lesion+GFP-MSCs, LPS lesion+GDNF-MSCs; n=10/group). Both cell groups received a unilateral intra-striatal transplant of either 200,000 GDNF-MSCs or 200,000 GFP-MSCs (as a control). One day post-transplantation, all rats received a unilateral intra-nigral infusion of LPS (10 μg in 2 μl sterile saline). Rats were sacrificed by transcardial perfusion-fixation and their brains were used for post mortem quantitative immunohistochemistry. Injection of LPS into the substantia nigra induced a pronounced local inflammatory response which resulted in 20% loss of nigrostriatal dopaminergic neurons and impaired contralateral motor function. Following transplantation of GDNF-MSCs to the striatum, dense areas of TH-positive staining directly proximal to the transplant site were observed. Most importantly, this effect was observed only in the GDNF-MSC transplanted group and not the GFP-MSC transplanted group demonstrating protection and/or sprouting of the dopaminergic terminals induced by the secreted GDNF. This study is the first to highlight the neurotrophic capability of GDNF in the inflammation-driven LPS model and, while future studies will endeavor to improve this approach by increasing cell survival, this work highlights the potential of GDNF delivery by ex vivo gene therapy using MSCs.
与胶质细胞源性神经营养因子（GDNF）递送方式相关的限制因素阻碍了其在帕金森病中作为神经保护剂的疗效。在体外基因治疗中，合适的细胞，如骨髓间充质干细胞（MSCs），在移植前经过基因工程改造以过度表达GDNF（GDNF-MSCs），这可能比直接向脑内输注神经营养因子更有益。此前，已在常用的帕金森病6-羟基多巴胺神经毒性模型中评估了GDNF-MSCs。然而，在本研究中，我们使用了一种新出现的帕金森病炎症模型（脂多糖（LPS）模型）来评估移植的GDNF-MSCs预防LPS诱导的神经炎症、神经退行性变和行为障碍的能力。本实验使用了30只雄性Sprague-Dawley大鼠。根据基线运动功能测试将大鼠性能匹配分为三组（仅LPS损伤组、LPS损伤+GFP-MSCs组、LPS损伤+GDNF-MSCs组；每组n = 10）。两个细胞组均接受了200,000个GDNF-MSCs或200,000个GFP-MSCs（作为对照）的单侧纹状体内移植。移植后一天，所有大鼠均接受了单侧黑质内LPS输注（10μg溶于2μl无菌盐水中）。通过心脏灌注固定处死大鼠，并将其大脑用于死后定量免疫组织化学分析。向黑质注射LPS诱发了明显的局部炎症反应，导致黑质纹状体多巴胺能神经元损失20%，对侧运动功能受损。将GDNF-MSCs移植到纹状体后，在移植部位紧邻处观察到TH阳性染色的密集区域。最重要的是，仅在GDNF-MSCs移植组中观察到这种效应，而在GFP-MSCs移植组中未观察到，这表明分泌的GDNF诱导了多巴胺能终末的保护和/或发芽。本研究首次突出了GDNF在炎症驱动的LPS模型中的神经营养能力，虽然未来的研究将致力于通过提高细胞存活率来改进这种方法，但这项工作突出了使用MSCs进行体外基因治疗递送GDNF的潜力。
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