Noninvasive gene transfer to the lung for systemic delivery of therapeutic proteins.
Noninvasive gene transfer to the lung for systemic delivery of therapeutic proteins.
复制标题
无创基因转移至肺部以全身递送治疗蛋白。
DOI：
10.1172/jci15780
复制
发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Wilson,JamesM
中科院分区：
文献类型：
--
作者：
Auricchio,Alberto;O'Connor,Erin;Weiner,Daniel;Gao,Guang-Ping;Hildinger,Markus;Wang,Lili;Calcedo,Roberto;Wilson,JamesM
本研究评估了基于腺相关病毒（aav）的载体的使用，将基因无创地传递到气道上皮细胞，作为实现治疗性蛋白全身给药的一种手段。我们将AAV载体经鼻传递给小鼠，其中编码细胞标记的相同AAV2基因组被包装在来自AAV1、2或5（分别为AAV2/1、AAV2/2或AAV2/5）的衣壳中。AAV2/5在气道和肺泡中的基因表达水平高于AAV2/1，而AAV2/2无法检测到。在肺特异性启动子（CC10）的控制下，将同一组编码分泌治疗性蛋白促红细胞生成素（Epo）的载体经鼻递送给小鼠，导致红细胞增多症，AAV2/5载体获得的血清促红细胞生成素水平最高。单次鼻内给药后，记录了150天的促红细胞生成素分泌。同样，经鼻给药aav2 /5 - cc10 -因子IX载体导致血友病因子IX缺陷小鼠血液中分泌功能性重组蛋白。此外，我们证明在首次递送相同载体5个月后，成功地将AAV2/5再给肺。总之，我们表明，只有当载体含有AAV5衣壳时，鼻内给药AAV载体才能有效地将基因转移到肺部，并且这种非侵入性给药途径可以在血液中持续分泌治疗蛋白。
This study evaluates the use of vectors based on adeno-associated viruses (AAVs) to noninvasively deliver genes to airway epithelial cells as a means for achieving systemic administration of therapeutic proteins. We intranasally delivered AAV vectors to mice in which the same AAV2 genome encoding a cellular marker was packaged in capsids from AAV1, 2, or 5 (AAV2/1, AAV2/2, or AAV2/5, respectively). Gene expression levels achieved in both airways and alveoli were higher with AAV2/5 than with AAV2/1 and were undetectable with AAV2/2. The same set of vectors encoding a secreted therapeutic protein, erythropoietin (Epo), under the control of a lung-specific promoter (CC10) was intranasally delivered to mice, resulting in polycythemia with the highest levels of serum Epo obtained with AAV2/5 vectors. After a single intranasal administration of this vector, secretion of Epo was documented for 150 days. Similarly, intranasal administration of an AAV2/5–CC10–factor IX vector resulted in secretion of functional recombinant protein in the bloodstream of hemophiliac, factor IX-deficient mice. In addition, we demonstrate successful readministration of AAV2/5 to the lung 5 months after the first delivery of the same vector. In conclusion, we show that intranasal administration of AAV vectors results in efficient gene transfer to the lung only when the vector contains the AAV5 capsid and that this noninvasive route of administration results in sustained secretion of therapeutic proteins in the bloodstream.