Radioisotopic imaging allows optimization of adenovirus lung deposition for cystic fibrosis gene therapy
Radioisotopic imaging allows optimization of adenovirus lung deposition for cystic fibrosis gene therapy
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DOI：
10.1089/104303401450915
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发表时间：
2001-01-01
影响因子：
4.2
通讯作者：
Pavirani, A
中科院分区：
文献类型：
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作者：
Lerondel, S;Le Pape, A;Pavirani, A
囊性纤维化是一种常见的遗传性疾病，由囊性纤维化跨膜传导调节基因突变引起。呼吸道转移CFTR基因是治疗或预防肺部病变的一种潜在策略，而肺部病变是导致发病率和死亡率的主要原因。在用于基因治疗的载体中，腺病毒已经显示出它们通过滴注或雾化将CFTR基因转移到呼吸道上皮细胞的能力。我们的目标是通过定量放射性同位素成像来表征雾化腺病毒的肺沉积，这是唯一一种能够在体内定量吸入药物的非侵入性技术。我们首先用发射伽马的放射性同位素~(99m)Tc标记了一个表达人CFTR的腺病毒，并确定了保持病毒生物活性的最佳标记条件。然后，我们用放射性气雾剂给狒狒注射，测定了99m标记的腺病毒在肺内的沉积，并比较了吸入3天和21天后CFTR转录本的表达，根据肺段的不同，载体编码的mRNA相对于内源性CFTRmRNA的表达从4%到22%不等。此外，我们还开发了一种使用二乙三胺五乙酸二乙三胺四乙酸二乙三胺的模型，该模型可以用来替代腺病毒，以确定载体的肺沉积轮廓。这项研究表明，核素扫描是实现呼吸道基因给药优化的一种有用的技术。
Cystic fibrosis is a common, heriditary disease resulting from mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Airway transfer of the CFTR gene is a potential strategy to treat or prevent the lung pathology that is the main cause of morbidity and mortality. Among the vectors used for gene therapy, adenoviruses have shown their ability to transfer the CFTR gene to respiratory epithelial cells, using either instillation or nebulization. Our objective was to characterize the lung deposition of aerosolized adenovirus by quantitative radioisotopic imaging, the only noninvasive technique allowing in vivo quantitation of inhaled drugs. We first labeled an adenovirus expressing human CFTR with the gamma -emitting radioisotope, technetium 99m (Tc-99m), and determined the best labeling conditions to allow preservation of virus bioactivity. We then administered the radioaerosol to baboons, determined lung regional deposition of Tc-99m-labeled adenovirus, and compared the expression of CFTR transcripts 3 and 21 days after inhalation, The expression of vector-encoded mRNA ranged from 4 to 22% with respect to the endogenous CFTR mRNA depending on the lung segments. Moreover, we have developed a model using Tc-99m-DTPA (diethylenetriamine pentaacetic acid), which can be used, as an alternative to adenovirus, to determine the profile of lung deposition of the vector. This study demonstrates that scintigraphy is a useful technique to achieve optimization of gene administration to the airways.