Zygmunt Deborah A, Xu Rui, Jia Ying, Ashbrook Anna, Menke Chelsea, Shao Guohong, Yoon Jung Hae, Hamilton Sonia, Pisharath Harshan, Bolon Brad, Martin Paul T
Center for Gene Therapy, Abigail Wexner Research Institute at Nationwide Children's Hospital, 700 Children's Drive, Columbus, OH, USA.
Animal Resources Core, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, OH, USA.
Mol Ther Methods Clin Dev. 2019 Oct 21;15:305-319. doi: 10.1016/j.omtm.2019.10.005. eCollection 2019 Dec 13.
rAAVrh74.MCK. is a surrogate gene therapy that inhibits muscular dystrophy in multiple animal models. Here, we report on a dose-response study of functional muscle expression as well as toxicity and biodistribution studies after systemic intravenous (i.v.) delivery of rAAVrh74.MCK.. A dose of 4.3 × 10vg/kg (measured with linear DNA standard) resulted in -induced glycosylation in the majority of skeletal myofibers throughout the body and in almost all cardiomyocytes, while several lower doses also showed significant muscle glycosylation. No adverse clinical signs or treatment-dependent changes in tissue or organ pathology were noted at 1 or 3 months post-treatment. Blood cell and serum enzyme chemistry measures in treated mice were all within the normal range except for alkaline phosphatase (ALP) activity, which was elevated in serum but not in tissues. Some anti-rAAVrh74 capsid T cell responses were noted at 4 weeks post-treatment, but all such responses were not present at 12 weeks. Using intramuscular delivery, -induced muscle glycosylation was increased in -deficient mice, which have a humanized sialoglycome, relative to wild-type mice, suggesting that use of mice may underestimate activity in human muscle. These data demonstrate safety and high transduction of muscles throughout the body plan with i.v. delivery of rAAVrh74.MCK..
重组腺相关病毒rh74.MCK.是一种替代基因疗法，可在多种动物模型中抑制肌肉萎缩症。在此，我们报告了rAAVrh74.MCK.经全身静脉注射后功能肌肉表达的剂量反应研究以及毒性和生物分布研究。剂量为4.3×10vg/kg（以线性DNA标准测量）时，可导致全身大多数骨骼肌纤维以及几乎所有心肌细胞出现诱导性糖基化，而几个较低剂量也显示出明显的肌肉糖基化。治疗后1个月或3个月未观察到不良临床体征或组织或器官病理学的治疗依赖性变化。除碱性磷酸酶（ALP）活性外，治疗小鼠的血细胞和血清酶化学指标均在正常范围内，碱性磷酸酶在血清中升高但在组织中未升高。治疗后4周观察到一些抗rAAVrh74衣壳T细胞反应，但在12周时所有此类反应均不存在。与野生型小鼠相比，在具有人源化唾液酸糖组的缺乏小鼠中，通过肌肉注射可增加诱导性肌肉糖基化，这表明使用小鼠可能会低估其在人肌肉中的活性。这些数据证明了通过静脉注射rAAVrh74.MCK.对全身肌肉进行安全且高效的转导。
