Adriaansen J, Perez P, Zheng C, Collins M T, Baum B J
Gene Transfer Section, Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892-1190, USA.
Hum Gene Ther. 2011 Jan;22(1):84-92. doi: 10.1089/hum.2010.097. Epub 2011 Jan 3.
Hypoparathyroidism is a hormone deficiency syndrome that leads to low blood calcium levels and for which current replacement therapy is inadequate. Gene transfer to salivary glands leads to safe and abundant secretion of therapeutic protein into either saliva or the bloodstream. We previously reported the successful transduction of rat submandibular glands with an adenoviral vector encoding human parathyroid hormone (Ad.hPTH), but unfortunately most of the hPTH was secreted into saliva. Because submandibular and parotid glands are morphologically and functionally different, we hypothesized that hPTH sorting might be different in parotid glands. After 2 days, the pattern of hPTH secretion from transduced parotid glands of intact rats was reversed from that of transduced submandibular glands, that is, most transgenic hPTH was detected in serum (5 × 10(10) viral particles per gland; the saliva-to-serum ratio of total hPTH secreted was 0.04). Vector copies were localized to the targeted parotid glands, with none detected in liver or spleen. Ad.hPTH next was administered to parotid glands of parathyroidectomized rats. Two days after delivery no hPTH was detectable in saliva, but high levels were found in serum, leading to normalization of serum calcium and a significant increase in the urinary phosphorus-to-creatinine ratio. This study demonstrates for the first time differential sorting of transgenic hPTH between submandibular and parotid glands, suggesting that hPTH may be a valuable model protein for understanding the molecular basis of transgenic secretory protein sorting in these exocrine glands. We also show the clinical potential of salivary gland hPTH gene therapy for patients with hypoparathyroidism.
甲状旁腺功能减退症是一种激素缺乏综合征,可导致血钙水平降低,且目前的替代疗法并不充分。将基因转移至唾液腺可使治疗性蛋白质安全且大量地分泌到唾液或血液中。我们之前报道了用编码人甲状旁腺激素(Ad.hPTH)的腺病毒载体成功转导大鼠下颌下腺,但遗憾的是,大部分hPTH分泌到了唾液中。由于下颌下腺和腮腺在形态和功能上存在差异,我们推测hPTH在腮腺中的分选可能不同。2天后,完整大鼠转导腮腺中hPTH的分泌模式与转导下颌下腺的模式相反,即大部分转基因hPTH在血清中被检测到(每腺体5×10¹⁰个病毒颗粒;分泌的总hPTH的唾液与血清比率为0.04)。载体拷贝定位于靶向腮腺,在肝脏或脾脏中未检测到。接下来将Ad.hPTH施用于甲状旁腺切除大鼠的腮腺。给药2天后,在唾液中未检测到hPTH,但在血清中发现了高水平的hPTH,导致血清钙正常化,尿磷与肌酐比率显著增加。本研究首次证明了转基因hPTH在下颌下腺和腮腺之间的差异分选,表明hPTH可能是理解这些外分泌腺中转基因分泌蛋白分选分子基础的有价值的模型蛋白。我们还展示了唾液腺hPTH基因治疗甲状旁腺功能减退症患者的临床潜力。