Stable delivery of physiologic levels of recombinant erythropoietin to the systemic circulation by intramuscular injection of replication-defective adenovirus.
Stable delivery of physiologic levels of recombinant erythropoietin to the systemic circulation by intramuscular injection of replication-defective adenovirus.
复制标题
通过肌内注射复制缺陷型腺病毒，将生理水平的重组促红细胞生成素稳定输送至体循环。
DOI：
10.1073/pnas.91.24.11557
复制
发表时间：
1994
影响因子：
11.1
通讯作者：
Leiden,JM
中科院分区：
文献类型：
--
作者：
Tripathy,SK;Goldwasser,E;Lu,MM;Barr,E;Leiden,JM
许多遗传性和获得性血清蛋白缺乏症，包括A型和B型血友病、糖尿病和红细胞生成素反应性贫血，目前通过反复皮下或静脉输注纯化或重组蛋白来治疗。开发体内基因转移方法以将生理水平的重组蛋白递送至体循环将代表治疗这些疾病的显著进步。在这里，我们描述了一个复制缺陷型腺病毒（AdEF 1hEpo）的构建含有人促红细胞生成素（hEpo）cDNA的转录控制下的细胞延伸因子1 α（EF 1 α）启动子和4F 2重链（4F 2 HC）增强子。将新生CD-1和成年SCID小鼠肌内（i.m.）用10（7）至10（9）空斑形成单位（pfu）的该病毒感染的小鼠显示出血清hEpo水平的显著剂量依赖性升高和血细胞比容增加，其在这些实验的4个月时间过程中是稳定的。肌内注射腺病毒仍然局限于注射部位，没有全身感染或局部炎症反应的证据。这些结果表明，i.m.注射重组复制缺陷型腺病毒载体可以作为治疗人血清蛋白缺乏的范例。
A number of inherited and acquired serum protein deficiencies including hemophilias A and B, diabetes mellitus, and the erythropoietin-responsive anemias are currently treated with repeated subcutaneous or intravenous infusions of purified or recombinant proteins. The development of an in vivo gene-transfer approach to deliver physiologic levels of recombinant proteins to the systemic circulation would represent a significant advance in the treatment of these disorders. Here we describe the construction of a replication-defective adenovirus (AdEF1hEpo) containing the human erythropoietin (hEpo) cDNA under the transcriptional control of the cellular elongation factor 1 alpha (EF1 alpha) promoter and the 4F2 heavy chain (4F2HC) enhancer. Neonatal CD-1 and adult SCID mice injected once intramuscularly (i.m.) with 10(7) to 10(9) plaque-forming units (pfu) of this virus displayed significant dose-dependent elevations of serum hEpo levels and increased hematocrits, which were stable over the 4-month time course of these experiments. Adenovirus injected i.m. remained localized at the site of injection and there was no evidence of either systemic infection or a localized inflammatory response. These results suggest that i.m. injection of recombinant replication-defective adenovirus vectors may serve as a paradigm for the treatment of human serum protein deficiencies.