Cationic, amphiphilic copolymer micelles as nucleic acid carriers for enhanced transfection in rat spinal cord.
Cationic, amphiphilic copolymer micelles as nucleic acid carriers for enhanced transfection in rat spinal cord.
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DOI：
10.1016/j.actbio.2016.02.013
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发表时间：
2016-04-15
影响因子：
9.7
通讯作者：
Lee JS
中科院分区：
文献类型：
--
作者：
Gwak SJ;Nice J;Zhang J;Green B;Macks C;Bae S;Webb K;Lee JS
由于成人中枢神经系统(CNS)再生能力有限，脊髓损伤通常会导致永久性的运动和感觉障碍。基于核酸的治疗是一种很有前途的策略，可以提供能够促进轴突再生的生物活性分子。支化聚乙烯亚胺(BPEI：25 kDa)是研究最广泛的非病毒载体之一，但由于其细胞毒性和在血清蛋白存在下的低转染率而限制了其临床应用。在本研究中，我们以约3：1的比例将低相对分子质量的PLGA(4 KDa)接枝到bPEI(25 KDa)上，合成了聚丙交酯-乙交酯-接枝-聚乙烯亚胺(PGP)阳离子两亲性共聚物。结果表明，在含10%血清的情况下，PGP胶束能有效地将pDNA和siRNA导入神经胶质瘤(C6)细胞、神经母细胞瘤(B35)细胞和原代E8鸡前脑神经元(CFN)，pDNA的转染率分别为58.8%、75.1%和8.1%。我们还表明，在活体大鼠脊髓中，PGP提供了比bPEI更高水平的转基因表达。与传统的bPEI相比，在有血清存在的情况下，PGP的体外转染率和对神经细胞的毒性降低相结合，这表明PGP可能是一种有前途的非病毒载体，用于治疗神经再生的核酸输送。
Spinal cord injury commonly leads to permanent motor and sensory deficits due to the limited regenerative capacity of the adult central nervous system (CNS). Nucleic acid-based therapy is a promising strategy to deliver bioactive molecules capable of promoting axonal regeneration. Branched polyethylenimine (bPEI: 25kDa) is one of the most widely studied nonviral vectors, but its clinical application has been limited due to its cytotoxicity and low transfection efficiency in the presence of serum proteins. In this study, we synthesized cationic amphiphilic copolymers, poly (lactide-co-glycolide)-graft-polyethylenimine (PgP), by grafting low molecular weight PLGA (4kDa) to bPEI (25kDa) at approximately a 3:1 ratio as an efficient nonviral vector. We show that PgP micelle is capable of efficiently transfecting plasmid DNA (pDNA) and siRNA in the presence of 10% serum in neuroglioma (C6) cells, neuroblastoma (B35) cells, and primary E8 chick forebrain neurons (CFN) with pDNA transfection efficiencies of 58.8%, 75.1 %, and 8.1 %, respectively. We also show that PgP provides high-level transgene expression in the rat spinal cord in vivo that is substantially greater than that attained with bPEI. The combination of improved transfection and reduced cytotoxicity in vitro in the presence of serum and in vivo transfection of neural cells relative to conventional bPEI suggests that PgP may be a promising nonviral vector for therapeutic nucleic acid delivery for neural regeneration.