Progress toward a nonviral gene therapy protocol for the treatment of anemia.
Progress toward a nonviral gene therapy protocol for the treatment of anemia.
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DOI：
10.1089/hum.2006.186
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发表时间：
2007-04
影响因子：
4.2
通讯作者：
M. G. Sebestyén;J. Hegge;M. Noble;D. Lewis;H. Herweijer;J. Wolff
中科院分区：
文献类型：
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作者：
M. G. Sebestyén;J. Hegge;M. Noble;D. Lewis;H. Herweijer;J. Wolff
贫血经常伴随慢性疾病，如进行性肾衰竭、获得性免疫缺陷综合征和癌症。目前使用各种重组人EPO蛋白制剂用促红细胞生成素（EPO）替代疗法治疗患者。虽然这种治疗是有效的，基因治疗可能是更经济和更方便的疾病的长期管理。本研究的目的是开发一种基于裸DNA的基因治疗方案，可以满足这一需求。流体动力学肢体静脉技术已被证明是一种有效和安全的程序，用于将裸质粒DNA（pDNA）输送到肢体的骨骼肌中。使用这种方法，我们解决了基于EPO的贫血基因治疗的主要挑战：在足以促进红细胞生成而不导致红细胞增多症的水平上保持稳定的长期表达。使用大鼠贫血模型，我们的研究结果提供了以下原理的证据：重复递送小剂量pDNA具有累加效应，并且可以逐渐导致贫血的纠正而不触发过度的造血。这种简单的方法提供了一种调节EPO表达的替代方法。在非人灵长类动物中，EPO表达也与注射的pDNA剂量成比例。此外，长期（超过450天）的表达后，提供恒河猴EPO cDNA下的肌肉特异性肌酸激酶（MCK）启动子的转录控制。总之，这些数据表明，重复递送小剂量的表达pDNA的EPO到骨骼肌中是一种有希望的、临床上可行的减轻贫血症状的方法。
Anemia frequently accompanies chronic diseases such as progressive renal failure, acquired immunodeficiency syndrome, and cancer. Patients are currently treated with erythropoietin (EPO) replacement therapy, using various recombinant human EPO protein formulations. Although this treatment is effective, gene therapy could be more economical and more convenient for the long-term management of the disease. The objective of this study was to develop a naked DNA-based gene therapy protocol that could fill this need. Hydrodynamic limb vein technology has been shown to be an effective and safe procedure for delivering naked plasmid DNA (pDNA) into the skeletal muscles of limbs. Using this method, we addressed the major challenge of an EPO-based gene therapy of anemia: maintaining stable, long-term expression at a level that sufficiently promotes erythropoiesis without leading to polycythemia. The results of our study, using a rat anemia model, provide proof of principle that repeated delivery of small pDNA doses has an additive effect and can gradually lead to the correction of anemia without triggering excessive hematopoiesis. This simple method provides an alternative approach for regulating EPO expression. EPO expression was also proportional to the injected pDNA dose in nonhuman primates. In addition, long-term (more than 450 days) expression was obtained after delivering rhesus EPO cDNA under the transcriptional control of the muscle-specific creatine kinase (MCK) promoter. In conclusion, these data suggest that the repeated delivery of small doses of EPO expressing pDNA into skeletal muscle is a promising, clinically viable approach to alleviate the symptoms of anemia.