Ataka Ken, Maruyama Hiroki, Neichi Tomohiro, Miyazaki Jun-ichi, Gejyo Fumitake
Division of Clinical Nephrology and Rheumatology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
Am J Nephrol. 2003 Sep-Oct;23(5):315-23. doi: 10.1159/000072913. Epub 2003 Aug 12.
We previously demonstrated that erythropoietin (Epo) expression increases in five-sixths nephrectomized rats, after muscle-targeted gene transfer by in vivo electroporation, using plasmid DNA expressing rat Epo (pCAGGS-Epo). Here, we apply this method to a rat model with severe anemia associated with chronic renal failure; these rats have hematocrit levels in the 30-35% range, similar to those in humans with end-stage renal disease.
Wistar rats were treated to produce adenine-induced uremia. The uremic rats were then treated with muscle-targeted gene transfer using pCAGGS-Epo. Some uremic rats died from chronic renal failure; one of these was dissected, and the kidneys were histologically examined. For the remaining rats, we measured body weight and blood pressure, and obtained blood samples regularly.
The uremic rats showed severe anemia, with hematocrit levels at 32.6 +/- 3.3%. Epo-gene transfer increased Epo expression and serum Epo levels, and also increased the hematocrit levels to 64.5 +/- 4.8%. The dose of pCAGGS-Epo used in this study did not induce severe hypertension.
Continuous Epo-gene expression improves the anemia associated with chronic renal failure, and without severe side effects. Our results support the potential use of gene electrotransfer for human gene therapy applications.
我们之前证明，通过体内电穿孔进行肌肉靶向基因转移，使用表达大鼠促红细胞生成素（Epo）的质粒DNA（pCAGGS-Epo）后，五分之六肾切除大鼠的Epo表达会增加。在此，我们将此方法应用于与慢性肾衰竭相关的严重贫血大鼠模型；这些大鼠的血细胞比容水平在30 - 35%范围内，与终末期肾病患者相似。
对Wistar大鼠进行处理以诱导腺嘌呤诱导的尿毒症。然后使用pCAGGS-Epo对尿毒症大鼠进行肌肉靶向基因转移治疗。一些尿毒症大鼠死于慢性肾衰竭；其中一只被解剖，对其肾脏进行组织学检查。对于其余大鼠，我们测量体重和血压，并定期采集血样。
尿毒症大鼠表现出严重贫血，血细胞比容水平为32.6±3.3%。Epo基因转移增加了Epo表达和血清Epo水平，同时也将血细胞比容水平提高到了64.5±4.8%。本研究中使用的pCAGGS-Epo剂量未诱导严重高血压。
持续的Epo基因表达可改善与慢性肾衰竭相关的贫血，且无严重副作用。我们的结果支持基因电转移在人类基因治疗应用中的潜在用途。
