High-efficiency Transduction of the Mouse Retina by Tyrosine-mutant AAV Serotype Vectors
High-efficiency Transduction of the Mouse Retina by Tyrosine-mutant AAV Serotype Vectors
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DOI：
10.1038/mt.2008.269
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发表时间：
2009-03-01
影响因子：
12.4
通讯作者：
Hauswirth, William W.
中科院分区：
文献类型：
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作者：
Petrs-Silva, Hilda;Dinculescu, Astra;Hauswirth, William W.
由腺相关病毒(AAVs)衍生的载体已经成为治疗许多遗传性眼病的重要基因输送工具。以前的研究已经确定，病毒衣壳在转基因表达的细胞趋向性和效率中起着至关重要的作用。最近，AAV2衣壳表面暴露的酪氨酸残基的磷酸化作用针对病毒颗粒进行泛素化和蛋白酶体介导的降解，而这些酪氨酸残基的突变在体外和体内都导致了高效的载体转导。由于酪氨酸残基在其他AAV血清型中高度保守，在本研究中，我们评估了含有AAV血清型2、8和9型表面暴露的衣壳酪氨酸残基点突变的载体的眼内转导特性。与野生型AAV突变体相比，其中几个新的AAV突变体在视网膜下或玻璃体内输送后在许多视网膜细胞中表现出强烈且广泛的转基因表达。我们首次展示了AAV8型或9型突变载体对神经节细胞层的有效转导，从而为AAV2以外的靶向这些细胞提供了额外的工具。这些增强型AAV载体在未来视网膜退行性变和眼部新生血管疾病的治疗应用中具有巨大的潜力。
Vectors derived from adeno-associated viruses (AAVs) have become important gene delivery tools for the treatment of many inherited ocular diseases in well-characterized animal models. Previous studies have determined that the viral capsid plays an essential role in the cellular tropism and efficiency of transgene expression. Recently, it was shown that phosphorylation of surface-exposed tyrosine residues from AAV2 capsid targets the viral particles for ubiquitination and proteasome-mediated degradation, and mutations of these tyrosine residues lead to highly efficient vector transduction in vitro and in vivo. Because the tyrosine residues are highly conserved in other AAV serotypes, in this study we evaluated the intraocular transduction characteristics of vectors containing point mutations in surface-exposed capsid tyrosine residues in AAV serotypes 2, 8, and 9. Several of these novel AAV mutants were found to display a strong and widespread transgene expression in many retinal cells after subretinal or intravitreal delivery compared with their wild-type counterparts. For the first time, we show efficient transduction of the ganglion cell layer by AAV serotype 8 or 9 mutant vectors, thus providing additional tools besides AAV2 for targeting these cells. These enhanced AAV vectors have a great potential for future therapeutic applications for retinal degenerations and ocular neovascular diseases.