RhoA knockdown by cationic amphiphilic copolymer/siRhoA polyplexes enhances axonal regeneration in rat spinal cord injury model.
RhoA knockdown by cationic amphiphilic copolymer/siRhoA polyplexes enhances axonal regeneration in rat spinal cord injury model.
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DOI：
10.1016/j.biomaterials.2017.01.003
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发表时间：
2017-03
期刊：
影响因子：
14
通讯作者：
Lee JS
中科院分区：
文献类型：
--
作者：
Gwak SJ;Macks C;Jeong DU;Kindy M;Lynn M;Webb K;Lee JS
脊髓损伤(SCI)会导致发育相关的外在微环境和内在神经元生化改变，从而限制可塑性和轴突再生，从而导致运动和感觉功能的永久性丧失。我们的长期目标是开发阳离子、两亲性共聚物(聚(丙交酯-乙交酯)-g-聚乙烯亚胺，PGP)，用于治疗性核酸(TNAS)和针对这些不同屏障的药物的组合递送。在这项研究中，我们评估了PGP递送针对RhoA的siRNA的能力，RhoA是由多种细胞外轴突再生抑制剂激活的关键信号通路。建立大鼠压迫性脊髓损伤模型后，将PGP/siRhoA复合体局部注射到损伤部位。与未经治疗的单纯损伤相比，PGP/siRhoA复合体在伤后4周内显著降低RhoA的mRNA和蛋白表达。损伤后4周的组织学分析显示，RhoA基因敲除后伴随着损伤部位细胞凋亡减少、空洞大小减少、星形胶质细胞增多和轴突再生增加。这些研究表明，PGP是一种有效的非病毒载体，将治疗性siRhoA转移到损伤的脊髓，有望成为TNA/药物联合治疗的一个有前途的平台。
Spinal cord injury (SCI) results in permanent loss of motor and sensory function due to developmentally-related and injured-induced changes in the extrinsic microenvironment and intrinsic neuronal biochemistry that limit plasticity and axonal regeneration. Our long term goal is to develop cationic, amphiphilic copolymers (poly (lactide-co-glycolide)-g-polyethylenimine, PgP) for combinatorial delivery of therapeutic nucleic acids (TNAs) and drugs targeting these different barriers. In this study, we evaluated the ability of PgP to deliver siRNA targeting RhoA, a critical signaling pathway activated by multiple extracellular inhibitors of axonal regeneration. After generation of rat compression SCI model, PgP/siRhoA polyplexes were locally injected into the lesion site. Relative to untreated injury only, PgP/siRhoA polyplexes significantly reduced RhoA mRNA and protein expression for up to 4 weeks post-injury. Histological analysis at 4 weeks post-injury showed that RhoA knockdown was accompanied by reduced apoptosis, cavity size, and astrogliosis and increased axonal regeneration within the lesion site. These studies demonstrate that PgP is an efficient non-viral delivery carrier for therapeutic siRhoA to the injured spinal cord and may be a promising platform for the development of combinatorial TNA/drug therapy.