Liu Mengping, Cook Erica, Dai Yanshan, Ehlert Erich, du Plessis Francois, Lubelski Jacek, Sleczka Bogdan G, Shipkova Petia, Li Zhuyin, Gamse Joshua, Gordon David, Adam Leonard P, Levesque Paul C, Banks Glen B
Bristol Myers Squibb, 3551 Lawrenceville Road, Princeton, NJ 08540, USA.
uniQure, Paasheuvelweg 25a, 1105 BP Amsterdam, the Netherlands.
Mol Ther Methods Clin Dev. 2025 Jan 17;33(1):101411. doi: 10.1016/j.omtm.2025.101411. eCollection 2025 Mar 13.
Safely achieving therapeutic expression levels with adeno-associated virus (AAV) gene therapy is a significant challenge for treating the large muscle mass in humans. Non-human primates (NHPs) provide a more accurate assessment of the feasibility of achieving an effective and safe dose than rodents. Here, we compared a single systemic administration of AAV5, AAV8, or AAV9 in NHPs, each packaging the C5-12-microdystrophin-FLAG expression cassette. At 1 month post-dose, we compared tissue vector genomes, mRNA, and microdystrophin-FLAG protein levels by meso-scale discovery-enzyme-linked immunosorbent assay, liquid chromatography-mass spectrometry, and immunofluorescence. The C5-12 promoter was highly selective for heart and skeletal muscles, when compared to off-target tissues such as peripheral blood mononuclear cells, lung, liver, and kidney. AAV8 led to higher levels of microdystrophin-FLAG mRNA and protein in the cardiac ventricles and skeletal muscles when compared to AAV5 or AAV9. The AAV8-microdystrophin-FLAG led to ∼20% of wild-type NHP dystrophin protein expression levels and was located on the sarcolemma of ∼40% of skeletal muscles fibers and ∼15% of left ventricular cardiomyocytes. Hematology, serum chemistry, and pathology were unremarkable. Thus, a systemic dose of ∼1.18 × 10 vector genomes/kg AAV8 is predicted to be safe and efficacious for treating Duchenne muscular dystrophy (DMD) but has significant room for improvement.
通过腺相关病毒（AAV）基因疗法安全地达到治疗表达水平，对于治疗人类大面积肌肉组织而言是一项重大挑战。与啮齿动物相比，非人灵长类动物（NHPs）能更准确地评估实现有效且安全剂量的可行性。在此，我们比较了在非人灵长类动物中单次全身给予AAV5、AAV8或AAV9的情况，每种病毒都包装了C5 - 12 - 微肌营养不良蛋白 - FLAG表达盒。给药后1个月，我们通过中尺度发现酶联免疫吸附测定、液相色谱 - 质谱联用和免疫荧光比较了组织载体基因组、mRNA和微肌营养不良蛋白 - FLAG蛋白水平。与外周血单核细胞、肺、肝和肾等脱靶组织相比，C5 - 12启动子对心脏和骨骼肌具有高度选择性。与AAV5或AAV9相比，AAV8在心室和骨骼肌中导致更高水平的微肌营养不良蛋白 - FLAG mRNA和蛋白。AAV8 - 微肌营养不良蛋白 - FLAG导致约20%的野生型非人灵长类动物肌营养不良蛋白表达水平，并且位于约40%的骨骼肌纤维和约15%的左心室心肌细胞的肌膜上。血液学、血清化学和病理学检查均无异常。因此，预计全身给予约1.18×10载体基因组/kg的AAV8对治疗杜氏肌营养不良症（DMD）是安全有效的，但仍有很大的改进空间。
