Comparative Study of Anti-hepatitis B Virus RNA Interference by Double-stranded Adeno-associated Virus Serotypes 7, 8, and 9
Comparative Study of Anti-hepatitis B Virus RNA Interference by Double-stranded Adeno-associated Virus Serotypes 7, 8, and 9
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DOI：
10.1038/mt.2008.245
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发表时间：
2009-02-01
影响因子：
12.4
通讯作者：
Tao, Mi-Hua
中科院分区：
文献类型：
--
作者：
Chen, Chun-Chi;Sun, Cheng-Pu;Tao, Mi-Hua
使用B肝炎病毒（HBV）转基因小鼠模型，我们以前表明，单剂量的双链腺相关病毒（dsAAV）载体血清型8携带一个小发夹RNA（shRNA）有效地减少HBV复制和基因表达，但效果随着时间的推移逐渐下降。在这份报告中，我们比较了dsAAV8与dsAAV7和dsAAV9，其他两种亲肝性AAV载体的抗HBV RNA干扰（RNAi）效果，并检查这些异源AAV载体的顺序使用是否可以延长抗HBV效果。我们的结果显示，由三种dsAAV载体中的每一种递送的shRNA在转基因小鼠中显著降低血清HBV滴度和肝脏HBV mRNA和DNA水平长达22周，其中dsAAV8具有最大的抑制效果，其次是dsAAV9和dsAAV7。dsAAV8的效力与肝脏中较高水平的载体DNA和抗HBV shRNA的存在相关。体内交叉给药实验表明，预先存在的抗AAV8抗体完全阻断了dsAAV8的抗HBV RNAi作用，但对dsAAV7和dsAAV9的效力没有影响。此外，我们证明了通过顺序使用dsAAV8和dsAAV9可以实现更长的抗HBV效果。这些结果表明，有效和持久的HBV抑制可以通过RNAi沉默效应的能力和不同AAV血清型的多次治疗的组合来实现。
Using a hepatitis B virus (HBV) transgenic mouse model, we previously showed that a single dose of double-stranded adeno-associated virus (dsAAV) vector serotype 8 carrying a small hairpin RNA (shRNA) effectively reduces HBV replication and gene expression, but the effect gradually decreases with time. In this report, we compared the anti-HBV RNA interference (RNAi) effect of dsAAV8 with those of dsAAV7 and dsAAV9, two other hepatotropic AAV vectors, and examined whether the sequential use of these heterologous AAV vectors could prolong the anti-HBV effect. Our results showed that shRNA delivered by each of the three dsAAV vectors profoundly reduced the serum HBV titer and liver HBV mRNA and DNA levels in the transgenic mice for up to 22 weeks, with dsAAV8 having the greatest inhibitory effect, followed by dsAAV9 and dsAAV7. The potency of dsAAV8 correlated with the presence of higher levels of vector DNA and anti-HBV shRNA in the liver. An in vivo cross-administration experiment showed that preexisting anti-AAV8 antibody completely blocked the anti-HBV RNAi effect of dsAAV8, but had no effect on the potency of dsAAV7 and dsAAV9. Moreover, we demonstrated that a longer anti-HBV effect could be achieved by the sequential use of dsAAV8 and dsAAV9. These results indicate that effective and persistent HBV suppression might be achieved by a combination of the power of RNAi silencing effect and multiple treatments with different AAV serotypes.