MicroRNA-mediated inhibition of transgene expression reduces dorsal root ganglion toxicity by AAV vectors in primates
MicroRNA-mediated inhibition of transgene expression reduces dorsal root ganglion toxicity by AAV vectors in primates
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DOI：
10.1126/scitranslmed.aba9188
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发表时间：
2020-11-11
影响因子：
17.1
通讯作者：
Wilson, James M.
中科院分区：
文献类型：
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作者：
Hordeaux, Juliette;Buza, Elizabeth L.;Wilson, James M.
通过血液或脑脊液将腺相关病毒（AAV）载体递送到非人灵长类动物（NHP）的中枢神经系统中与背根神经节（DRG）毒性相关。常规的免疫抑制方案不能防止这种毒性，可能是因为它可能是由高转导速率引起的，高转导速率又会由于靶细胞中转基因产物的过量而引起细胞应激。为了验证这一假设并开发一种消除DRG毒性的方法，我们利用microRNA（miR）183复合物的内源性表达，其主要限于DRG神经元，以特异性下调这些细胞中的转基因表达。我们将miR183的序列靶标引入载体基因组中相应转基因信使RNA的3'非翻译区内，并将载体注射到NHP的小脑延髓池中。施用未修饰的AAV载体导致靶组织的稳健转导和DRG神经元中的毒性。与免疫系统活性不介导这种神经元毒性的提议一致，我们发现类固醇给药在缓解这种病理学方面无效。然而，在载体中包括miR183靶标降低了DRG神经元中的转基因表达和毒性，而不影响灵长类动物脑中其他地方的转导。这种方法可能有助于减少DRG毒性和相关的发病率，并应促进许多中枢神经系统疾病的基于AAV的基因疗法的发展。
Delivering adeno-associated virus (AAV) vectors into the central nervous system of nonhuman primates (NHPs) via the blood or cerebral spinal fluid is associated with dorsal root ganglion (DRG) toxicity. Conventional immunesuppression regimens do not prevent this toxicity, possibly because it may be caused by high transduction rates, which can, in turn, cause cellular stress due to an overabundance of the transgene product in target cells. To test this hypothesis and develop an approach to eliminate DRG toxicity, we exploited endogenous expression of microRNA (miR) 183 complex, which is largely restricted to DRG neurons, to specifically down-regulate transgene expression in these cells. We introduced sequence targets for miR183 into the vector genome within the 3' untranslated region of the corresponding transgene messenger RNA and injected vectors into the cisterna magna of NHPs. Administration of unmodified AAV vectors resulted in robust transduction of target tissues and toxicity in DRG neurons. Consistent with the proposal that immune system activity does not mediate this neuronal toxicity, we found that steroid administration was ineffective in alleviating this pathology. However, including miR183 targets in the vectors reduced transgene expression in, and toxicity of, DRG neurons without affecting transduction elsewhere in the primate's brain. This approach might be useful in reducing DRG toxicity and the associated morbidity and should facilitate the development of AAV-based gene therapies for many central nervous system diseases.