Development of optimized AAV3 serotype vectors: mechanism of high-efficiency transduction of human liver cancer cells.
Development of optimized AAV3 serotype vectors: mechanism of high-efficiency transduction of human liver cancer cells.
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我们最近的研究表明，在10种不同的常用AAV血清型中，AAV3载体非常有效地抑制人肝癌细胞，因为这些细胞表达高水平的人肝细胞生长因子受体（hHGFR），并且AAV3利用hHGFR作为病毒进入的细胞共受体。在这份报告中，我们提供了进一步的证据表明，细胞外以及细胞内的激酶结构域的hHGFR参与AAV3载体进入和AAV3介导的转基因表达。我们还记载了AAV 3载体被靶向用于通过宿主细胞蛋白酶体机制降解，并且AAV 3衣壳上表面暴露的酪氨酸（Y）至苯丙氨酸（F）残基的定点诱变显著提高了Y701F、Y705F和Y731F突变体AAV 3载体的转导效率。Y705+731F双突变体载体在体外肝癌细胞中的转导效率显著高于各单突变体。在免疫缺陷小鼠异种移植模型中，直接肿瘤内注射AAV3载体也导致体内人肝肿瘤细胞的高效转导。我们还在此记录了优化的酪氨酸突变体AAV3载体在体内肿瘤内和尾静脉注射后导致转导效率增加。优化的酪氨酸突变型AAV3血清型载体含有促凋亡基因，应证明是有用的潜在的人类肝癌的基因治疗。
Our recent studies have revealed that among the 10 different commonly used AAV serotypes, AAV3 vectors transduce human liver cancer cells extremely efficiently because these cells express high levels of human hepatocyte growth factor receptor (hHGFR), and AAV3 utilizes hHGFR as a cellular co-receptor for viral entry. In this report, we provide further evidence that both extracellular as well as intracellular kinase domains of hHGFR are involved in AAV3 vector entry and AAV3-mediated transgene expression. We also document that AAV3 vectors are targeted for degradation by the host cell proteasome machinery, and that site-directed mutagenesis of surface exposed tyrosine (Y) to phenylalanine (F) residues on AAV3 capsids significantly improves the transduction efficiency of Y701F, Y705F and Y731F mutant AAV3 vectors. The transduction efficiency of the Y705+731F double-mutant vector is significantly higher than each of the single-mutants in liver cancer cells in vitro. In immuno-deficient mouse xenograft models, direct intra-tumor injection of AAV3 vectors also led to high-efficiency transduction of human liver tumor cells in vivo. We also document here that the optimized tyrosine-mutant AAV3 vectors lead to increased transduction efficiency following both intra-tumor and tail-vein injections in vivo. The optimized tyrosine-mutant AAV3 serotype vectors containing pro-apoptotic genes should prove useful for the potential gene therapy of human liver cancers.
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影响因子：
3.2
作者：
Chen, Tina T. -L.;Rakheja, Dinesh;Tomlinson, Gail E.
通讯作者：
Tomlinson, Gail E.
影响因子：
5.4
作者：
Hansen, J;Qing, K;Srivastava, A
通讯作者：
Srivastava, A
影响因子：
5.4
作者：
Ferrari, FK;Samulski, T;Samulski, RJ
通讯作者：
Samulski, RJ
影响因子：
5.4
作者：
Douar, AM;Poulard, K;Danos, O
通讯作者：
Danos, O
DOI：
10.1073/pnas.182412299
发表时间：
2002-09-03
影响因子：
11.1
作者：
Gao, GP;Alvira, MR;Wilson, JM
通讯作者：
Wilson, JM