Sustained and complete phenotype correction of hemophilia B mice following intramuscular injection of AAV1 serotype vectors
Sustained and complete phenotype correction of hemophilia B mice following intramuscular injection of AAV1 serotype vectors
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DOI：
10.1006/mthe.2001.0449
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发表时间：
2001-09-01
影响因子：
12.4
通讯作者：
Walsh, CE
中科院分区：
文献类型：
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作者：
Chao, HJ;Monahan, PE;Walsh, CE
我们以前曾报道，直接肌肉注射非血清2型AAV载体，特别是AAV1(AAV1)，可导致免疫缺陷小鼠犬F9异常表达。在这里，我们测试了AAV1-F9载体在基因工程血友病小鼠中持续表达和纠正因子IX(FIX)缺陷的能力。肌肉注射AAV1-F9后，重组AAV1-F9小鼠血浆中犬F9的含量是注射AAV2-F9的100-1000倍。通过激活的部分凝血活酶时间评估凝血活性，证实了循环犬固定物确实具有功能。此外，通过尾巴夹击的表型校正试验表明，所有AAV1-F9处理的动物都存活了下来，而幼稚的小鼠和AAV2处理的血友病B小鼠中有50%未能存活。对于AAV2处理的动物，需要环磷酰胺(CTX)来抑制抗犬FIX抗体的形成，而对于AAV1-F9处理的动物，环磷酰胺(CTX)的注射是必要的。这种免疫原性的差异进一步强调了血清型特异性载体的用处。最后，我们报告了使用AAV1-F9对血友病表型的纠正是完全和持久的(超过8个月)，这一结果强调了继续探索替代AAV血清型载体的价值。
We previously reported that direct intramuscular injection of non-serotype-2 AAV vectors, especially AAV serotype 1 (AAV1), resulted in expression of supranormal levels of canine F9 in immunodeficient mice. Here we test the ability of the AAV1-F9 vector to deliver sustained expression and correction of factor IX (FIX) deficiency in genetically engineered hemophilic mice. Intramuscular injection of AAV1-F9 resulted in 100-1000 times more canine F9 in plasma of recombinant AAV1-F9 mice compared with injection of AAV2-F9. Assessment of clotting activity by activated partial thromboplastin time confirmed that circulating canine FIX was indeed functional. Moreover, phenotypic correction assayed by tail clip challenge resulted in survival of all AAV1-F9 treated animals, in contrast to naive mice and 50 % of AAV2-treated hemophilia B mice, which failed to survive. Administration of cyclophosphamide (CTX) was required to suppress formation of anti-canine FIX antibodies for AAV2-treated animals, whereas it was dispensable for those treated with AAV1-F9. This difference in immunogenicity further emphasizes the usefulness of serotype-specific vectors. Finally, we report that correction of the hemophilia phenotype using AAV1-F9 was complete and persistent (over 8 months), a result that underscores the value of continued exploration of alternative AAV serotype vectors.