Robust Long-term Transduction of Common Marmoset Neuromuscular Tissue With rAAV1 and rAAV9.
Robust Long-term Transduction of Common Marmoset Neuromuscular Tissue With rAAV1 and rAAV9.
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DOI：
10.1038/mtna.2013.21
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发表时间：
2013-05-28
期刊：
影响因子：
--
通讯作者：
Okada T
中科院分区：
文献类型：
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作者：
Okada H;Ishibashi H;Hayashita-Kinoh H;Chiyo T;Nitahara-Kasahara Y;Baba Y;Watanabe S;Takeda S;Okada T
重组腺相关病毒（rAAV）介导的转导的概况显示每个AAV血清型的种间差异。在啮齿类动物中通常观察到稳健的长期转基因表达，而在具有更高级免疫系统的动物中观察到不充分的转导。非人类灵长类动物，包括普通的绒猴，可以为神经肌肉疾病提供合适的模型，因为它们的大脑功能更高，生理上与人类相似。通过rAAV介导的转导在非人灵长类动物的神经肌肉组织中诱导病理的策略是有希望的;然而，在绒猴中尚未阐明rAAV转导的转基因表达模式。在这项研究中，用rAAV 9转导成年绒猴骨骼肌导致稳健和持续增强的绿色荧光蛋白（EGFP）表达，其与肌纤维类型无关，尽管识别出淋巴细胞浸润。全身性rAAV注射到妊娠绒猴中导致经胎盘胎儿转导。令人惊讶的是，将rAAV 1和rAAV 9腹膜内注射到新生绒猴中导致全身转导和持续的转基因表达，而没有淋巴细胞浸润。分别用rAAV 1和rAAV 9有效地转导了骨骼肌和心肌。有趣的是，rAAV 9转导导致胼胝体轴突中强烈的EGFP信号传导。这些rAAV的转导方案将有助于研究神经肌肉组织中的基因功能和开发基因治疗策略。
Profiles of recombinant adeno-associated virus (rAAV)-mediated transduction show interspecies differences for each AAV serotype. Robust long-term transgene expression is generally observed in rodents, whereas insufficient transduction is seen in animals with more advanced immune systems. Non-human primates, including the common marmoset, could provide appropriate models for neuromuscular diseases because of their higher brain functions and physiological resemblance to humans. Strategies to induce pathologies in the neuromuscular tissues of non-human primates by rAAV-mediated transduction are promising; however, transgene expression patterns with rAAV transduction have not been elucidated in marmosets. In this study, transduction of adult marmoset skeletal muscle with rAAV9 led to robust and persistent enhanced green fluorescent protein (EGFP) expression that was independent of the muscle fiber type, although lymphocyte infiltration was recognized. Systemic rAAV injection into pregnant marmosets led to transplacental fetal transduction. Surprisingly, the intraperitoneal injection of rAAV1 and rAAV9 into the neonatal marmoset resulted in systemic transduction and persistent transgene expression without lymphocyte infiltration. Skeletal and cardiac muscle were effectively transduced with rAAV1 and rAAV9, respectively. Interestingly, rAAV9 transduction led to intense EGFP signaling in the axons of the corpus callosum. These transduction protocols with rAAV will be useful for investigating gene functions in the neuromuscular tissues and developing gene therapy strategies.