Immune-evasive gene switch enables regulated delivery of chondroitinase after spinal cord injury.
Immune-evasive gene switch enables regulated delivery of chondroitinase after spinal cord injury.
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DOI：
10.1093/brain/awy158
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发表时间：
2018-08-01
期刊：
影响因子：
--
通讯作者：
Bradbury EJ
中科院分区：
文献类型：
--
作者：
Burnside ER;De Winter F;Didangelos A;James ND;Andreica EC;Layard-Horsfall H;Muir EM;Verhaagen J;Bradbury EJ
软骨素酶是一种很有前途的脊髓损伤后功能恢复的临床前治疗方法。伯恩赛德等人设计了一种免疫逃避载体系统，该系统允许在开/关开关的控制下递送软骨素酶以调节表达。开启软骨素酶有助于脊髓损伤大鼠恢复伸手和抓握运动。 软骨素酶ABC是一种很有前途的临床前治疗，通过降解细胞外基质抑制剂促进CNS损伤后的功能性神经可塑性。通过病毒载体转基因递送，可以将软骨素酶ABC有效递送至受损的哺乳动物脊髓。这种方法极大地调节损伤病理学并恢复感觉运动功能。然而，这种疗法的临床开发受到缺乏对软骨素酶基因表达施加控制的能力的限制。先前的实验性基因调控平台可能与已知在脊髓损伤后发生的非解决性适应性免疫反应不相容。因此，在这里，我们应用了一种新的免疫逃避双载体系统，其中软骨素酶基因是在强力霉素诱导的调节开关，利用嵌合反式激活因子，旨在逃避T细胞的识别。使用这种新的载体系统，我们证明了严格的时间控制软骨素酶ABC基因的表达，有效地消除治疗后，去除强力霉素。这使得短期和长期的基因治疗模式在治疗成年大鼠的临床相关的颈部水平挫伤的比较。我们发现，短暂治疗（2.5周）足以促进感觉轴突传导和阶梯行走性能的改善。然而，在需要熟练的伸手和抓握的任务中，只有长期治疗（8周）才能显著改善功能，大鼠能够准确地抓握和取回糖丸。熟练的手功能的后期出现表明增强的神经可塑性和连通性，并与脊髓灰质中vGlut 1+神经支配的密度增加相关，特别是在损伤上方和下方的层III-IV中。因此，我们的新型基因治疗系统提供了一种实验工具，以研究细胞外基质消化的时间效应，以及产生一个更安全的软骨素酶基因治疗策略的一个令人鼓舞的步骤，长期管理，其中增加神经可塑性和恢复下降运动控制。这项临床前研究可能会对四肢瘫痪患者产生重大影响，对他们来说，手功能的恢复是独立的重要决定因素，并支持软骨素酶基因治疗的持续发展，以临床应用于治疗脊髓损伤。
Chondroitinase is a promising preclinical therapy for restoring function after spinal cord injury. Burnside et al. engineer an immune-evasive vector system that allows delivery of chondroitinase under the control of an on/off switch to regulate expression. Switching on chondroitinase helps spinal-injured rats to recover reaching and grasping movements. Chondroitinase ABC is a promising preclinical therapy that promotes functional neuroplasticity after CNS injury by degrading extracellular matrix inhibitors. Efficient delivery of chondroitinase ABC to the injured mammalian spinal cord can be achieved by viral vector transgene delivery. This approach dramatically modulates injury pathology and restores sensorimotor functions. However, clinical development of this therapy is limited by a lack of ability to exert control over chondroitinase gene expression. Prior experimental gene regulation platforms are likely to be incompatible with the non-resolving adaptive immune response known to occur following spinal cord injury. Therefore, here we apply a novel immune-evasive dual vector system, in which the chondroitinase gene is under a doxycycline inducible regulatory switch, utilizing a chimeric transactivator designed to evade T cell recognition. Using this novel vector system, we demonstrate tight temporal control of chondroitinase ABC gene expression, effectively removing treatment upon removal of doxycycline. This enables a comparison of short and long-term gene therapy paradigms in the treatment of clinically-relevant cervical level contusion injuries in adult rats. We reveal that transient treatment (2.5 weeks) is sufficient to promote improvement in sensory axon conduction and ladder walking performance. However, in tasks requiring skilled reaching and grasping, only long term treatment (8 weeks) leads to significantly improved function, with rats able to accurately grasp and retrieve sugar pellets. The late emergence of skilled hand function indicates enhanced neuroplasticity and connectivity and correlates with increased density of vGlut1+ innervation in spinal cord grey matter, particularly in lamina III–IV above and below the injury. Thus, our novel gene therapy system provides an experimental tool to study temporal effects of extracellular matrix digestion as well as an encouraging step towards generating a safer chondroitinase gene therapy strategy, longer term administration of which increases neuroplasticity and recovery of descending motor control. This preclinical study could have a significant impact for tetraplegic individuals, for whom recovery of hand function is an important determinant of independence, and supports the ongoing development of chondroitinase gene therapy towards clinical application for the treatment of spinal cord injury.
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DOI：
10.1056/nejmoa1700554
发表时间：
2017-10-26
期刊：
The New England journal of medicine
影响因子：
--
作者：
Eichler F;Duncan C;Musolino PL;Orchard PJ;De Oliveira S;Thrasher AJ;Armant M;Dansereau C;Lund TC;Miller WP;Raymond GV;Sankar R;Shah AJ;Sevin C;Gaspar HB;Gissen P;Amartino H;Bratkovic D;Smith NJC;Paker AM;Shamir E;O'Meara T;Davidson D;Aubourg P;Williams DA
通讯作者：
Williams DA
影响因子：
3.3
作者：
Du Beau, A.;Shrestha, S. Shakya;Maxwell, D. J.
通讯作者：
Maxwell, D. J.
影响因子：
5.3
作者：
ChenKinon;Marsh, Barnaby C.;Ichiyama, Ronaldo M.
通讯作者：
Ichiyama, Ronaldo M.
影响因子：
3.4
作者：
Fouad K;Bennett DJ;Vavrek R;Blesch A
通讯作者：
Blesch A
影响因子：
2.5
作者：
Alvarez, FJ;Villalba, RM;Schneider, SP
通讯作者：
Schneider, SP