Voutetakis Antonis, Kok Marc R, Zheng Changyu, Bossis Ioannis, Wang Jianghua, Cotrim Ana P, Marracino Natanya, Goldsmith Corinne M, Chiorini John A, Loh Y Peng, Nieman Lynnette K, Baum Bruce J
Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892, USA.
Proc Natl Acad Sci U S A. 2004 Mar 2;101(9):3053-8. doi: 10.1073/pnas.0400136101. Epub 2004 Feb 20.
The use of critical-for-life organs (e.g., liver or lung) for systemic gene therapeutics can lead to serious safety concerns. To circumvent such issues, we have considered salivary glands (SGs) as an alternative gene therapeutics target tissue. Given the high secretory abilities of SGs, we hypothesized that administration of low doses of recombinant adeno-associated virus (AAV) vectors would allow for therapeutic levels of transgene-encoded secretory proteins in the bloodstream. We administered 10(9) particles of an AAV vector encoding human erythropoietin (hEPO) directly to individual mouse submandibular SGs. Serum hEPO reached maximum levels 8-12 weeks after gene delivery and remained relatively stable for 54 weeks (longest time studied). Hematocrit levels were similarly increased. Moreover, these effects proved to be vector dose-dependent, and even a dosage as low as 10(8) particles per animal led to significant increases in hEPO and hematocrit levels. Vector DNA was detected only within the targeted SGs, and levels of AAV copies within SGs were highly correlated with serum hEPO levels (r = 0.98). These results show that SGs appear to be promising targets with potential clinical applicability for systemic gene therapeutics.
将维持生命关键的器官（如肝脏或肺）用于全身基因治疗可能会引发严重的安全问题。为规避此类问题，我们已将唾液腺（SGs）视为基因治疗的替代靶组织。鉴于唾液腺的高分泌能力，我们推测给予低剂量的重组腺相关病毒（AAV）载体将使转基因编码的分泌蛋白在血液中达到治疗水平。我们将编码人促红细胞生成素（hEPO）的AAV载体的10⁹个颗粒直接注射到单个小鼠的下颌下唾液腺中。基因递送后8 - 12周血清hEPO达到最高水平，并在54周（研究的最长时间）内保持相对稳定。血细胞比容水平也同样升高。此外，这些效应被证明是载体剂量依赖性的，甚至每只动物低至10⁸个颗粒的剂量也会导致hEPO和血细胞比容水平显著升高。仅在靶向的唾液腺中检测到载体DNA，并且唾液腺内AAV拷贝数与血清hEPO水平高度相关（r = 0.98）。这些结果表明，唾液腺似乎是全身基因治疗具有潜在临床适用性的有前景的靶标。
