Department of Biopharmaceutics, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan; Laboratory of Biopharmaceutics, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
Department of Biopharmaceutics, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto 607-8414, Japan; Laboratory of Biopharmaceutics, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
J Control Release. 2018 Apr 10;275:78-84. doi: 10.1016/j.jconrel.2018.02.018. Epub 2018 Feb 16.
Though cell transplantation is becoming an attractive therapeutic method, uncontrolled cell proliferation or overexpression of cellular functions could cause adverse effects. These unfavorable outcomes could be avoided by regulating the proliferation or functioning of transplanted cells. In this study, we used a combination of the herpes simplex virus thymidine kinase (HSVtk) gene, a suicide gene, and ganciclovir (GCV) to control the proliferation and functioning of insulin-secreting cells after transplantation in diabetic mice. Mouse pancreatic β cell line MIN6 cells were selected as insulin-secreting cells for transfection with the HSVtk gene to obtain MIN6/HSVtk cells. Proliferation of MIN6/HSVtk cells was suppressed by GCV in a concentration-dependent manner; 0.25 μg/mL GCV maintained a constant number of MIN6/HSVtk cells for at least 16 days. MIN6 or MIN6/HSVtk cells were then transplanted to streptozotocin-induced diabetic mice. Mice transplanted with MIN6 cells exhibited hypoglycemia irrespective of GCV administration. In contrast, normal (around 150 mg/dL) blood glucose levels were maintained in mice transplanted with MIN6/HSVtk cells by a daily administration of 50 mg/kg of GCV. These results indicate that controlling the proliferation and functioning of HSVtk gene-expressing cells by GCV could greatly improve the usefulness and safety of cell-based therapy.
虽然细胞移植正成为一种有吸引力的治疗方法，但不受控制的细胞增殖或过度表达细胞功能可能会导致不良影响。通过调节移植细胞的增殖或功能，可以避免这些不利的结果。在这项研究中，我们使用单纯疱疹病毒胸苷激酶（HSVtk）基因、自杀基因和更昔洛韦（GCV）的组合来控制糖尿病小鼠移植后胰岛素分泌细胞的增殖和功能。选择小鼠胰岛β细胞系 MIN6 细胞作为转染 HSVtk 基因的胰岛素分泌细胞，以获得 MIN6/HSVtk 细胞。MIN6/HSVtk 细胞的增殖在浓度依赖性方式下被 GCV 抑制；0.25μg/mL GCV 至少能维持 MIN6/HSVtk 细胞的数量稳定 16 天以上。然后将 MIN6 或 MIN6/HSVtk 细胞移植到链脲佐菌素诱导的糖尿病小鼠中。无论是否给予 GCV，移植 MIN6 细胞的小鼠均表现出低血糖。相比之下，每天给予 50mg/kg GCV 可使移植 MIN6/HSVtk 细胞的小鼠的血糖水平维持在正常水平（约 150mg/dL）。这些结果表明，通过 GCV 控制 HSVtk 基因表达细胞的增殖和功能，可以大大提高细胞治疗的有效性和安全性。
