Triple Vectors Expand AAV Transfer Capacity in the Retina.
Triple Vectors Expand AAV Transfer Capacity in the Retina.
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DOI：
10.1016/j.ymthe.2017.11.019
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发表时间：
2018-02-07
期刊：
影响因子：
--
通讯作者：
Auricchio A
中科院分区：
文献类型：
--
作者：
Maddalena A;Tornabene P;Tiberi P;Minopoli R;Manfredi A;Mutarelli M;Rossi S;Simonelli F;Naggert JK;Cacchiarelli D;Auricchio A
用腺相关病毒（AAV）载体进行视网膜基因转移对于治疗遗传性视网膜变性（IRD）具有很大的希望。AAV的一个限制是其约5kb的转移能力，使用双重AAV载体可以将其扩增至约9kb。这种策略仍然不足以治疗IRD，如Usher综合征1D型或Alström综合征I型（ALMS），分别由于CDH23或ALMS 1突变。为了克服这一限制，我们产生了三重AAV载体，其最大转移容量为约14 kb。三重AAV转导后的转录组学分析显示预期的全长产物沿着许多异常转录物。然而，只有全长转录本在体内有效翻译。我们还表明，约4%的小鼠光感受器被三重AAV载体转导，并显示重组ALMS 1的正确定位。低光感受器转导水平可能证明我们在ALMS小鼠模型的视网膜中观察到的适度和短暂的改善是合理的。然而，在猪视网膜中由三重AAV载体介导的转导水平达到用单载体观察到的水平的40%，这预示着进一步提高三重AAV载体在视网膜中的效率。Maddalena等人显示腺相关病毒（AAV）载体的有限DNA转移能力可以用三重AAV载体扩展至14kb。作者在体外以及小鼠和猪视网膜中显示了三重AAV介导的报告基因和大治疗基因的表达。
Retinal gene transfer with adeno-associated viral (AAV) vectors holds great promise for the treatment of inherited retinal degenerations (IRDs). One limit of AAV is its transfer capacity of about 5 kb, which can be expanded to about 9 kb, using dual AAV vectors. This strategy would still not suffice for treatment of IRDs such as Usher syndrome type 1D or Alström syndrome type I (ALMS) due to mutations in CDH23 or ALMS1, respectively. To overcome this limitation, we generated triple AAV vectors, with a maximal transfer capacity of about 14 kb. Transcriptomic analysis following triple AAV transduction showed the expected full-length products along a number of aberrant transcripts. However, only the full-length transcripts are efficiently translated in vivo. We additionally showed that approximately 4% of mouse photoreceptors are transduced by triple AAV vectors and showed correct localization of recombinant ALMS1. The low-photoreceptor transduction levels might justify the modest and transient improvement we observe in the retina of a mouse model of ALMS. However, the levels of transduction mediated by triple AAV vectors in pig retina reached 40% of those observed with single vectors, and this bodes well for further improving the efficiency of triple AAV vectors in the retina. Maddalena et al. show that the limited DNA transfer capacity of adeno-associated viral (AAV) vectors can be expanded up to 14 kb with triple AAV vectors. The authors show triple AAV-mediated expression of both reporter and large therapeutic genes in vitro and in mouse and pig retina.
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影响因子：
--
作者：
Doria, Monica;Ferrara, Antonella;Auricchio, Alberto
通讯作者：
Auricchio, Alberto
影响因子：
17.8
作者：
Berger, Wolfgang;Kloeckener-Gruissem, Barbara;Neidhardt, John
通讯作者：
Neidhardt, John
影响因子：
3.7
作者：
Colella P;Sommella A;Marrocco E;Di Vicino U;Polishchuk E;Garcia Garrido M;Seeliger MW;Polishchuk R;Auricchio A
通讯作者：
Auricchio A
影响因子：
5.4
作者：
Allocca, Mariacarmela;Mussolino, Claudio;Auricchio, Alberto
通讯作者：
Auricchio, Alberto
影响因子：
4.2
作者：
Boye, Shannon E.;Alexander, John J.;Gamlin, Paul D.
通讯作者：
Gamlin, Paul D.