Safety and in vivo expression of a GNE-transgene: a novel treatment approach for hereditary inclusion body myopathy-2.

Anagha P Phadke, Chris Jay, Salina J Chen, Courtney Haddock, Zhaohui Wang, Yang Yu, Derek Nemunaitis, Gregory Nemunaitis, Nancy S Templeton, Neil Senzer, Phillip B Maples, Alex W Tong, John Nemunaitis
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引用次数: 10

Abstract

Hereditary inclusion body myopathy-2 (HIBM2) is an adult-onset, muscular disease caused by mutations in the GNE gene. HIBM2-associated GNE mutations causing hyposialyation have been proposed to contribute to reduced muscle function in patients with HIBM2, though the exact cause of this disease is unknown. In the current studies we examined pre-clinical in vivo toxicity, and expression of the plasmid-based, CMV driven wild-type GNE plasmid vector. The plasmid vector was injected intramuscularly (IM) or systemically (IV) into BALB/c mice, following encapsulation in a cationic liposome (DOTAP:Cholesterol). Single IM injections of the GNE-lipoplex at 40 microg did not produce overt toxicity or deaths, indicating that the no observable adverse effect level (NOAEL) dose for IM injection was >or=40 microg. Single intravenous (IV) infusion of GNE-lipoplex was lethal in 33% of animals at 100 microg dose, with a small proportion of animals in the 40 microg cohort demonstrating transient toxicity. Thus the NOAEL dose by the IV route was greater than 10 microg and less than or equal to 40 microg. Real-time RT-qPCR analysis demonstrated recombinant human GNE mRNA expression in 100% of muscle tissues that received IM injection of 40 microg GNE-lipoplex, at 2 weeks. These results indicate that GNE-lipoplex gene transfer is safe and can produce durable transgene expression in treated muscles. Our findings support future exploration of the clinical efficacy of GNE-lipoplex for experimental gene therapy of HIBM2.

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转基因基因的安全性和体内表达:遗传性包涵体肌病-2的新治疗方法。
遗传性包涵体肌病-2 (HIBM2)是一种由GNE基因突变引起的成人发病的肌肉疾病。已提出HIBM2相关的GNE突变导致低水平,有助于HIBM2患者肌肉功能降低,尽管这种疾病的确切原因尚不清楚。在目前的研究中,我们检测了临床前体内毒性,以及基于质粒的、CMV驱动的野生型GNE质粒载体的表达。将质粒载体包封在阳离子脂质体(DOTAP:胆固醇)中,肌注(IM)或全身(IV)注射到BALB/c小鼠中。单次注射40微克的gne -脂质体未产生明显的毒性或死亡,表明注射40微克的IM的无观察到不良反应水平(NOAEL)剂量>或=40微克。单次静脉(IV)输注GNE-lipoplex在100微克剂量下对33%的动物致命,在40微克队列中有一小部分动物表现出短暂毒性。由此可见,静脉给药NOAEL剂量大于10 μ g,小于等于40 μ g。实时RT-qPCR分析显示,在IM注射40微克GNE-脂质体2周后,100%的肌肉组织中表达了重组人GNE mRNA。这些结果表明GNE-lipoplex基因转移是安全的,并且可以在处理过的肌肉中产生持久的转基因表达。我们的研究结果支持了GNE-lipoplex用于实验性HIBM2基因治疗的临床疗效的进一步探索。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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