Comparison of the transduction efficiency of tyrosine-mutant adeno-associated virus serotype vectors in kidney.
Comparison of the transduction efficiency of tyrosine-mutant adeno-associated virus serotype vectors in kidney.
复制标题
比较肾脏中与酪氨酸突变腺相关病毒血清型载体的转导效率的比较。
DOI：
10.1111/1440-1681.12037
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发表时间：
2013-01
影响因子：
2.9
通讯作者：
Raizada MK
中科院分区：
文献类型：
--
作者：
Qi YF;Li QH;Shenoy V;Zingler M;Jun JY;Verma A;Katovich MJ;Raizada MK
1.基因疗法在治疗肾脏疾病方面具有独特的潜力。然而，通过病毒载体有效转导大量肾细胞一直难以实现。先前的研究表明腺相关病毒（AAV）可以以可变的和次优的效率感染肾细胞。由于新的和创新的AAV突变体，我们比较了它们在转导大鼠肾脏的功效。2.我们比较了携带绿色荧光蛋白（GFP）报告基因的五种类型的AAV突变体（AAV 2 muttriple、AAV 2 sixtuple、AAV 8 mut 447、AAV 8 mut 733和AAV 9 mut 446）。采用压力显微注射技术，将1.5× 10 ~（11）个AAV突变体载体基因组（vg）或3个剂量的AAV_2（6倍）注射入肾皮质。显微注射的方法还没有被用于AAV介导的肾基因转移到目前为止。缓慢和持续的显微注射使得能够将病毒载体连续施用到肾皮质，并限制对肾脏的任何损伤，因为玻璃微量移液管的尖端非常小。3.注射后三周，收集肾脏并评估GFP表达。在所研究的各种突变的AAV血清型中，仅AAV 2六倍体在肾组织中显示出稳健的GFP表达。AAV 2六重血清型似乎是优先靶向肾小管上皮细胞的有效基因转移载体。AAV 2 sixtuple和显微注射技术的结合是未来肾脏疾病治疗的关键。
1. Gene therapy has a distinct potential to treat kidney diseases. However, the efficient transduction of a significant number of renal cells by viral vectors has been difficult to accomplish. Previous studies have indicated that adeno-associated virus (AAV) can transduce renal cells with variable and suboptimal efficiency. Since new and innovative mutants of AAV are now available, we compared their efficacy in transducing rat kidneys. 2. We compared five types of AAV mutants (AAV2 muttriple, AAV2 sextuple, AAV8 mut447, AAV8 mut733, and AAV9 mut446) that carried a green fluorescence protein (GFP) reporter gene. A pressure microinjection technique was used to inject either 1.5×1011 vector genome (vg) of AAV mutants or three dose of AAV2 sextuple into the renal cortex. The microinjection approach has not been utilized in AAV-mediated renal gene transfer thus far. Slow and sustained microinjection enables continuous administration of the viral vector to the kidney cortex and limits any damage to the kidney, as the tip of a glass micropipette is very small. 3. Three weeks after injection, the kidneys were collected and evaluated for GFP expression. Among the various mutated AAV serotypes studied, only AAV2 sextuple showed a robust GFP expression in renal tissue. The AAV2 sextuple serotype appears to be an efficient gene transfer vector to preferentially target the renal tubular epithelial cells. A combination of the AAV2 sextuple and the microinjection technique holds the key to the future of therapeutic treatments for kidney diseases.
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影响因子：
12.4
作者：
Petrs-Silva, Hilda;Dinculescu, Astra;Hauswirth, William W.
通讯作者：
Hauswirth, William W.
影响因子：
3.2
作者：
Loghman-Adham, M;Soto, CE;Sotelo-Avila, C
通讯作者：
Sotelo-Avila, C
影响因子：
5.1
作者：
Zolotukhin, S;Byrne, BJ;Muzyczka, N
通讯作者：
Muzyczka, N
影响因子：
5.4
作者：
Yang, GS;Schmidt, M;Davidson, BL
通讯作者：
Davidson, BL
影响因子：
3.5
作者：
Qi, Yanfei;Liu, Xuan;Katovich, Michael J.
通讯作者：
Katovich, Michael J.