Transduction of Neural Precursor Cells with TAT-Heat Shock Protein 70 Chaperone: Therapeutic Potential Against Ischemic Stroke after Intrastriatal and Systemic Transplantation
Transduction of Neural Precursor Cells with TAT-Heat Shock Protein 70 Chaperone: Therapeutic Potential Against Ischemic Stroke after Intrastriatal and Systemic Transplantation
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DOI：
10.1002/stem.1098
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发表时间：
2012-06-01
期刊：
影响因子：
5.2
通讯作者：
Baehr, Mathias
中科院分区：
文献类型：
--
作者：
Doeppner, Thorsten R.;Ewert, Tobias A. S.;Baehr, Mathias
针对脑缺血的新的治疗概念集中在基于细胞的治疗上，以克服溶栓治疗的一些副作用。然而，由于对最佳细胞移植途径的了解有限，以及移植细胞的存活率较低，基于细胞的治疗受到阻碍。因此，我们将成年绿色荧光蛋白阳性神经前体细胞移植到小鼠中风后6小时的静脉(全身)或纹状体内(大脑内)。为了提高神经干细胞的存活能力，在移植前用TAT-热休克蛋白70(Hsp70)体外转导细胞，然后系统分析脑损伤和依赖于细胞传递途径的潜在机制。携带TAT-HSP70的神经干细胞转导后，脑内移植的神经干细胞数量增加，而全身移植后的神经干细胞数量没有增加。尽管全身注射天然或转导的神经前体可以产生持续的神经保护和诱导神经功能恢复，但只有转导TAT-HSP70的神经前体可以防止脑内注射后的继发性神经元变性，这与增强的功能结局有关。此外，TAT-HSP70转导的神经干细胞脑内移植促进了脑缺血后的神经再生，并在体内诱导了持续高水平的脑源性神经营养因子、胶质细胞源性神经营养因子和血管内皮生长因子。脑细胞移植后的神经保护与存活的神经前体细胞的数量相关。相反，全身给药通过稳定血脑屏障来介导急性神经保护，同时伴随着基质金属蛋白酶9活性的降低和活性氧物种的减少。我们的发现暗示了两种不同的作用机制，即脑内移植和系统移植的鼻咽癌，这表明全身鼻咽癌给药可能更适合翻译卒中的概念，不需要体外操作鼻咽癌来诱导长期的神经保护。STEM CELLS2012；30：12971310
Novel therapeutic concepts against cerebral ischemia focus on cell-based therapies in order to overcome some of the side effects of thrombolytic therapy. However, cell-based therapies are hampered because of restricted understanding regarding optimal cell transplantation routes and due to low survival rates of grafted cells. We therefore transplanted adult green fluorescence protein positive neural precursor cells (NPCs) either intravenously (systemic) or intrastriatally (intracerebrally) 6 hours after stroke in mice. To enhance survival of NPCs, cells were in vitro protein-transduced with TAT-heat shock protein 70 (Hsp70) before transplantation followed by a systematic analysis of brain injury and underlying mechanisms depending on cell delivery routes. Transduction of NPCs with TAT-Hsp70 resulted in increased intracerebral numbers of grafted NPCs after intracerebral but not after systemic transplantation. Whereas systemic delivery of either native or transduced NPCs yielded sustained neuroprotection and induced neurological recovery, only TAT-Hsp70-transduced NPCs prevented secondary neuronal degeneration after intracerebral delivery that was associated with enhanced functional outcome. Furthermore, intracerebral transplantation of TAT-Hsp70-transduced NPCs enhanced postischemic neurogenesis and induced sustained high levels of brain-derived neurotrophic factor, glial cell line-derived neurotrophic factor, and vascular endothelial growth factor in vivo. Neuroprotection after intracerebral cell delivery correlated with the amount of surviving NPCs. On the contrary, systemic delivery of NPCs mediated acute neuroprotection via stabilization of the blood-brain-barrier, concomitant with reduced activation of matrix metalloprotease 9 and decreased formation of reactive oxygen species. Our findings imply two different mechanisms of action of intracerebrally and systemically transplanted NPCs, indicating that systemic NPC delivery might be more feasible for translational stroke concepts, lacking a need of in vitro manipulation of NPCs to induce long-term neuroprotection. STEM CELLS2012;30:12971310