经基因修饰以过表达 BDNF、CNTF、GDNF、NGF、NT3 或 VEGF 的外周神经自体移植物中轴突再生的比较形态学、电生理学和功能分析。
A comparative morphological, electrophysiological and functional analysis of axon regeneration through peripheral nerve autografts genetically modified to overexpress BDNF, CNTF, GDNF, NGF, NT3 or VEGF.
机构信息
Department of Neuroregeneration, Netherlands Institute for Neuroscience, Royal Netherlands Academy of Arts and Sciences, Meibergdreef 47, 1105BA Amsterdam, The Netherlands; Department of Neurosurgery, Leiden University Medical Center, Albinusdreef 2, 2333ZA Leiden, The Netherlands.
Department of Neuroregeneration, Netherlands Institute for Neuroscience, Royal Netherlands Academy of Arts and Sciences, Meibergdreef 47, 1105BA Amsterdam, The Netherlands; Department of Clinical and Biological Sciences, Neuroscience Institute of the Cavalieri Ottolenghi Foundation, University of Turin, Turin, Italy.
出版信息
Exp Neurol. 2014 Nov;261:578-93. doi: 10.1016/j.expneurol.2014.08.002. Epub 2014 Aug 12.
The clinical outcome of microsurgical repair of an injured peripheral nerve with an autograft is suboptimal. A key question addressed here is: can axon regeneration through an autograft be further improved? In this article the impact of six neurotrophic factors (BDNF, CNTF, GDNF, NGF, NT3 or VEGF) on axon regeneration was compared after delivery to a 1cm long nerve autograft by gene therapy. To distinguish between early and late effects, regeneration was assessed at 2 and 20weeks post-surgery by histological, electrophysiological and functional analysis. BDNF, GDNF and NGF exhibited a spectrum of effects, including early stimulatory effects on axons entering the autograft and excessive axon growth and Schwann cell proliferation at 20weeks post-surgery. Persistent expression of these factors in autografts interfered with target cell reinnervation and functional recovery in a modality specific way. Autografts overexpressing VEGF displayed hypervascularization, while grafts transduced with CNTF and NT3 were indistinguishable from control grafts. These three factors did not have detectable pro-regenerative effects. In conclusion, autograft-based repair combined with gene therapy for three of the six growth factors investigated (BDNF, GDNF, NGF) showed considerable promise since these factors enhanced modality specific axon outgrowth in autografts. The remarkable and selective effects of BDNF, GDNF and NGF on motor or sensory regeneration will be exploited in future experiments that aim to carefully regulate their temporal and spatial expression since this has the potential to overcome the adverse effects on long-distance regeneration observed after uncontrolled delivery.
自体移植修复周围神经损伤的临床效果并不理想。这里要解决的一个关键问题是:通过自体移植进行轴突再生能否进一步改善?本文通过基因治疗,比较了 6 种神经营养因子(BDNF、CNTF、GDNF、NGF、NT3 或 VEGF)在移植 1cm 长神经自体移植物后的轴突再生的影响。为了区分早期和晚期的影响,在手术后 2 周和 20 周通过组织学、电生理学和功能分析来评估再生情况。BDNF、GDNF 和 NGF 表现出一系列的影响,包括对进入自体移植物的轴突的早期刺激作用,以及在手术后 20 周时的过度轴突生长和施万细胞增殖。这些因子在自体移植物中的持续表达以特定的方式干扰了靶细胞的再神经支配和功能恢复。过表达 VEGF 的自体移植物表现出血管过度生成,而转导 CNTF 和 NT3 的移植物与对照移植物没有区别。这三种因子没有可检测的促再生作用。总之,结合基因治疗,在研究的 6 种生长因子中的 3 种(BDNF、GDNF、NGF)中,自体移植修复显示出了很大的潜力,因为这些因子增强了自体移植物中特定模态的轴突生长。BDNF、GDNF 和 NGF 对运动或感觉再生的显著和选择性作用将在未来的实验中得到利用,这些实验旨在仔细调节它们的时空表达,因为这有可能克服在不受控制的递送后观察到的对长距离再生的不利影响。