中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (3): 550-560.doi: 10.4103/1673-5374.293150

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

硬膜外电刺激结合离体三基因疗法治疗脊髓损伤

  

  • 出版日期:2021-03-15 发布日期:2020-12-17
  • 基金资助:

    这项研究得到了俄罗斯科学基金会（RRI）和喀山联邦大学竞争发展计划的支持

Combination of epidural electrical stimulation with ex vivo triple gene therapy for spinal cord injury: a proof of principle study

Filip Olegovich Fadeev1, Farid Vagizovich Bashirov1, Vahe Arshaluysovich Markosyan1, Andrey Alexandrovich Izmailov1, Tatyana Vyacheslavovna Povysheva1, #br# Mikhail Evgenyevich Sokolov1, Maxim Sergeevich Kuznetsov1, #br# Anton Alexandrovich Eremeev2, Ilnur Ildusovich Salafutdinov2, #br# Albert Anatolyevich Rizvanov2, Hyun Joon Lee3, 4, *, Rustem Robertovich Islamov1, *#br#   

  1. 1 Department of Medical Biology and Genetics, Kazan State Medical University, Kazan, Russia;  2 Institute of Fundamental Medicine and Biology, Kazan [Volga Region] Federal University, Kazan, Russia;  3 Department of Neurology, University of Mississippi Medical Center, Jackson, MS, USA;  4 Research Service, G.V. (Sonny) Montgomery VA Medical Center, Jackson, MS, USA
  • Online:2021-03-15 Published:2020-12-17
  • Contact: Hyun Joon Lee, PhD, hlee@umc.edu; Rustem Robertovich Islamov, MD, Dr.Sci., rustem.islamov@gmail.com.
  • Supported by:
    This study was supported by the grant of Russian Science Foundation, No. 16-15-00010 (to RRI).  AAR was supported by the Russian Government Program of Competitive Growth of Kazan Federal University.

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摘要：

目前在临床上尚无公认有效的治疗脊髓损伤的基因和干细胞的治疗方法。作者既往的研究表明，基因工程化的人脐带血单个核细胞（UCB-MC）的移植可产生3种重组治疗分子，包括血管内皮生长因子（VEGF），胶质细胞源性神经营养因子（GDNF）和神经细胞黏附分子（ NCAM）可促进大鼠和小型猪脊髓创伤性损伤后的形态和功能恢复。为了解UCB-MC介导的上述3种分子基因疗法与硬膜外电刺激联合用于SCI治疗的效果。实验于大鼠中度脊髓挫伤后4h，行脊髓鞘内注射表达重组基因VEGF，GDNF，NCAM的UCB-MC。伤后3d给予脊髓C5损伤部位上方（用于刺激与前肢功能有关的颈部神经网络）和L2损伤部位下方（用于激活中央模式发生器）硬膜外刺激。可见联合治疗一定程度改善了脊髓损伤大鼠膝关节运动功能，减少了脊髓组织细胞凋亡，保留了脊髓灰质和白质完整性，并增强脊髓组织热休克蛋白和突触蛋白的表达。该研究阐释了硬膜外电刺激与离体三基因疗法（VEGF，GDNF和NCAM）作为治疗SCI潜在疗法的作用机制。

https://orcid.org/0000-0002-6985-0690 (Hyun Joon Lee);

https://orcid.org/0000-0002-6632-4636 (Rustem Robertovich Islamov)

关键词:

脊髓损伤, 硬膜外电刺激, 基因疗法, 人脐带血单个核细胞, 腺病毒载体, 血管内皮生长因子, 胶质细胞源性神经营养因子, 神经细胞黏附分子

Abstract: Despite emerging contemporary biotechnological methods such as gene- and stem cell-based therapy, there are no clinically established therapeutic strategies for neural regeneration after spinal cord injury. Our previous studies have demonstrated that transplantation of genetically engineered human umbilical cord blood mononuclear cells producing three recombinant therapeutic molecules, including vascular endothelial growth factor (VEGF), glial cell-line derived neurotrophic factor (GDNF), and neural cell adhesion molecule (NCAM) can improve morpho-functional recovery of injured spinal cord in rats and mini-pigs. To investigate the efficacy of human umbilical cord blood mononuclear cells-mediated triple-gene therapy combined with epidural electrical stimulation in the treatment of spinal cord injury, in this study, rats with moderate spinal cord contusion injury were intrathecally infused with human umbilical cord blood mononuclear cells expressing recombinant genes VEGF165, GDNF, NCAM1 at 4 hours after spinal cord injury. Three days after injury, epidural stimulations were given simultaneously above the lesion site at C5 (to stimulate the cervical network related to forelimb functions) and below the lesion site at L2 (to activate the central pattern generators) every other day for 4 weeks.  Rats subjected to the combined treatment showed a limited functional improvement of the knee joint, high preservation of muscle fiber area in tibialis anterior muscle and increased H/M ratio in gastrocnemius muscle 30 days after spinal cord injury. However, beneficial cellular outcomes such as reduced apoptosis and increased sparing of the gray and white matters, and enhanced expression of heat shock and synaptic proteins were found in rats with spinal cord injury subjected to the combined epidural electrical stimulation with gene therapy. This study presents the first proof of principle study of combination of the multisite epidural electrical stimulation with ex vivo triple gene therapy (VEGF, GDNF and NCAM) for treatment of spinal cord injury in rat models. The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee (approval No. 2.20.02.18) on February 20, 2018.  

Key words:

adenoviral vector, epidural electrical stimulation, gene therapy, glial cell-line derived neurotrophic factor, human umbilical cord blood mononuclear cell, neural cell adhesion molecule, spinal cord injury, vascular endothelial growth factor