Persistent expression of human clotting factor IX from mouse liver after intravenous injection of adeno-associated virus vectors
Persistent expression of human clotting factor IX from mouse liver after intravenous injection of adeno-associated virus vectors
复制标题
DOI：
10.1073/pnas.94.4.1426
复制
发表时间：
1997-02-18
影响因子：
11.1
通讯作者：
Miller, AD
中科院分区：
文献类型：
--
作者：
Koeberl, DD;Alexander, IE;Miller, AD
我们以前发现，腺相关病毒（AAV）载体在细胞培养中的基因转导可以刺激超过100倍的治疗与影响DNA代谢的药物，如辐射或拓扑异构酶抑制剂的靶细胞。在这里，我们表明，以前的γ射线照射增加了小鼠肝脏的转导率高达900倍，拓扑异构酶抑制剂依托泊苷增加了约20倍的转导。通过直接注射肝脏或通过经由尾静脉注射的全身递送获得相似的肝转导速率。在全身递送后，肝细胞比其他细胞更有效地转导，并且在一次载体注射后，所有肝细胞的高达3%可以被转导。需要存在污染许多AAV载体制剂的野生型AAV以观察对γ-照射的完全响应。在γ-照射后用编码人凝血因子IX的AAV载体注射小鼠导致在5个月的观察期内合成低水平的人凝血因子IX。这些研究显示了通过AAV载体进行肝脏靶向基因转导用于治疗各种血液学或代谢疾病的潜力。
We previously found that gene transduction by adeno-associated virus (AAV) vectors in cell culture can be stimulated over 100-fold by treatment of the target cells with agents that affect DNA metabolism, such as irradiation or topoisomerase inhibitors. Here we show that previous gamma-irradiation increased the transduction rate in mouse liver by up to 900-fold, and the topoisomerase inhibitor etoposide increased transduction by about 20-fold. Similar rates of hepatic transduction were obtained by direct injection of the liver or by systemic delivery via tail vein injection, Hepatocytes were much more efficiently transduced than other cells after systemic delivery, and up to 3% of all hepatocytes could be transduced after one vector injection, The presence of wildtype AAV, which contaminates many AAV vector preparations, was required to observe a full response to gamma-irradiation. Injection of mice with AAV vectors encoding human clotting factor IX after gamma-irradiation resulted in synthesis of low levels of human clotting factor IX for the 5-month period of observation. These studies show the potential of targeted gene transduction of the liver by AAV vectors for treatment of various hematological or metabolic diseases.