Kwon Joon Hyun, Lee Young Mok, Cho Jun-Ho, Kim Goo-Young, Anduaga Javier, Starost Matthew F, Mansfield Brian C, Chou Janice Y
Section on Cellular Differentiation, Eunice Kennedy Shriver National Institute of Child Health and Human Development.
Division of Veterinary Resources, National Institutes of Health, Bethesda, MD 20892, USA.
Hum Mol Genet. 2017 Nov 15;26(22):4395-4405. doi: 10.1093/hmg/ddx325.
Glycogen storage disease type-Ib (GSD-Ib), deficient in the glucose-6-phosphate transporter (G6PT), is characterized by impaired glucose homeostasis, myeloid dysfunction, and long-term risk of hepatocellular adenoma (HCA). We examined the efficacy of G6PT gene therapy in G6pt-/- mice using recombinant adeno-associated virus (rAAV) vectors, directed by either the G6PC or the G6PT promoter/enhancer. Both vectors corrected hepatic G6PT deficiency in murine GSD-Ib but the G6PC promoter/enhancer was more efficacious. Over a 78-week study, using dose titration of the rAAV vectors, we showed that G6pt-/- mice expressing 3-62% of normal hepatic G6PT activity exhibited a normalized liver phenotype. Two of the 12 mice expressing < 6% of normal hepatic G6PT activity developed HCA. All treated mice were leaner and more sensitive to insulin than wild-type mice. Mice expressing 3-22% of normal hepatic G6PT activity exhibited higher insulin sensitivity than mice expressing 44-62%. The levels of insulin sensitivity correlated with the magnitudes of hepatic carbohydrate response element binding protein signaling activation. In summary, we established the threshold of hepatic G6PT activity required to prevent tumor formation and showed that mice expressing 3-62% of normal hepatic G6PT activity maintained glucose homeostasis and were protected against age-related obesity and insulin resistance.
I型糖原贮积病(GSD-Ib)因葡萄糖-6-磷酸转运体(G6PT)缺乏,其特征为葡萄糖稳态受损、髓系功能障碍以及肝细胞腺瘤(HCA)的长期风险。我们使用由G6PC或G6PT启动子/增强子指导的重组腺相关病毒(rAAV)载体,检测了G6PT基因疗法对G6pt-/-小鼠的疗效。两种载体均纠正了小鼠GSD-Ib中的肝脏G6PT缺乏,但G6PC启动子/增强子更有效。在一项为期78周的研究中,通过对rAAV载体进行剂量滴定,我们发现,表达正常肝脏G6PT活性3%-62%的G6pt-/-小鼠表现出正常的肝脏表型。12只表达<正常肝脏G6PT活性6%的小鼠中有2只发生了HCA。所有接受治疗的小鼠都比野生型小鼠更瘦,对胰岛素更敏感。表达正常肝脏G6PT活性3%-22%的小鼠比表达44%-62%的小鼠表现出更高的胰岛素敏感性。胰岛素敏感性水平与肝脏碳水化合物反应元件结合蛋白信号激活的程度相关。总之,我们确定了预防肿瘤形成所需的肝脏G6PT活性阈值,并表明表达正常肝脏G6PT活性3%-62%的小鼠维持了葡萄糖稳态,并免受与年龄相关的肥胖和胰岛素抵抗的影响。