Adriaansen J, Perez P, Goldsmith C M, Zheng C, Baum B J
Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892-1190, USA.
Hum Gene Ther. 2008 Oct;19(10):1021-8. doi: 10.1089/hum.2008.079.
Gene transfer to salivary glands leads to abundant secretion of transgenic protein into either saliva or the bloodstream. This indicates significant clinical potential, depending on the route of sorting. The aim of this study was to probe the sorting characteristics of human parathyroid hormone (hPTH) in two animal models for salivary gland gene transfer. PTH is a key hormone regulating calcium levels in the blood. A recombinant serotype 5 adenoviral vector carrying the hPTH cDNA was administered to the submandibular glands of mice and rats. Two days after delivery, high levels of hPTH were found in the serum of mice, leading to elevated serum calcium levels. Only low amounts of hPTH were found in the saliva. Two days after vector infusion into rats, a massive secretion of hPTH was measured in saliva, with little secretion into serum. Confocal microscopy showed hPTH in the glands, localized basolaterally in mice and apically in rats. Submandibular gland transduction was effective and the produced hPTH was biologically active in vivo. Whereas hPTH sorted toward the basolateral side in mice, in rats hPTH was secreted mainly at the apical side. These results indicate that the interaction between hPTH and the cell sorting machinery is different between mouse and rat salivary glands. Detailed studies in these two species should result in a better understanding of cellular control of transgenic secretory protein sorting in this tissue.
基因转移至唾液腺可导致转基因蛋白大量分泌到唾液或血液中。这显示出巨大的临床潜力,具体取决于分选途径。本研究的目的是在两种用于唾液腺基因转移的动物模型中探究人甲状旁腺激素(hPTH)的分选特性。甲状旁腺激素是调节血液中钙水平的关键激素。将携带hPTH cDNA的重组5型腺病毒载体注射到小鼠和大鼠的下颌下腺。注射两天后,在小鼠血清中发现高水平的hPTH,导致血清钙水平升高。而在唾液中仅发现少量hPTH。将载体注入大鼠两天后,检测到唾液中有大量hPTH分泌,而分泌到血清中的量很少。共聚焦显微镜检查显示腺体中有hPTH,在小鼠中位于基底外侧,在大鼠中位于顶端。下颌下腺转导有效,所产生的hPTH在体内具有生物活性。hPTH在小鼠中向基底外侧分选,而在大鼠中hPTH主要在顶端分泌。这些结果表明,hPTH与细胞分选机制之间的相互作用在小鼠和大鼠唾液腺中有所不同。对这两个物种进行详细研究应有助于更好地理解该组织中转基因分泌蛋白分选的细胞调控机制。