School of Pharmacy, East China University of Science and Technology, Shanghai, China.
State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai, China.
Gene Ther. 2024 Jan;31(1-2):19-30. doi: 10.1038/s41434-023-00411-3. Epub 2023 Jul 27.
Adeno-associated virus (AAV) vectors have been successfully used to deliver genes for treating rare diseases. However, the systemic administration of high AAV vector doses triggers several adverse effects, including immune response, the asymptomatic elevation of liver transaminase levels, and complement activation. Thus, improving AAV transduction and reducing AAV dosage for treatment is necessary. Recently, we found that a phosphodiesterase-5 inhibitor significantly promoted AAV9 transduction in vitro by regulating the caveolae and macropinocytosis pathways. When AAV9-Gaussian luciferase (AAV9-Gluc) and AAV9-green fluorescent protein (AAV9-GFP) were injected intravenously into mice pre-treated with sildenafil, the expressions of Gluc in the plasma and GFP in muscle tissues significantly increased (P < 0.05). Sildenafil also improved Evans blue permeation in tissues. Additionally, we found that sildenafil promoted Treg proliferation, inhibited B-cell activation, and decreased anti-AAV9 IgG levels (P < 0.05). Furthermore, sildenafil significantly promoted Duchenne muscular dystrophy gene therapy efficacy using AAV9 in mdx mice; it increased micro-dystrophin gene expression, forelimb grip strength, and time spent on the rotarod test, decreased serum creatine kinase levels, and ameliorated histopathology by improving muscle cell morphology and reducing fibrosis (P < 0.05). These results show that sildenafil significantly improved AAV transduction, suppressed the levels of anti-AAV9 IgG, and enhanced the efficacy of gene therapy.
腺相关病毒 (AAV) 载体已成功用于递送基因以治疗罕见疾病。然而,高剂量的 AAV 载体全身给药会引发多种不良反应,包括免疫反应、肝转氨酶水平的无症状升高和补体激活。因此,提高 AAV 的转导效率并降低治疗剂量是必要的。最近,我们发现磷酸二酯酶-5 抑制剂通过调节小窝和巨胞饮途径,显著促进了 AAV9 的体外转导。当预先用西地那非处理的小鼠静脉注射 AAV9-高斯荧光素酶 (AAV9-Gluc) 和 AAV9-绿色荧光蛋白 (AAV9-GFP) 时,血浆中 Gluc 的表达和肌肉组织中 GFP 的表达显著增加(P<0.05)。西地那非也改善了组织中的 Evans 蓝渗透。此外,我们发现西地那非促进了 Treg 的增殖,抑制了 B 细胞的激活,并降低了抗 AAV9 IgG 水平(P<0.05)。此外,西地那非在 mdx 小鼠中使用 AAV9 显著促进了杜氏肌营养不良症的基因治疗疗效;它增加了微肌营养不良蛋白基因的表达、前肢握力和在旋转棒测试上的时间,降低了血清肌酸激酶水平,并通过改善肌肉细胞形态和减少纤维化来改善组织病理学(P<0.05)。这些结果表明,西地那非显著提高了 AAV 的转导效率,抑制了抗 AAV9 IgG 的水平,并增强了基因治疗的疗效。