Evaluation of photoreceptor transduction efficacy of capsid-modified AAV vectors following intravitreal and subretinal delivery in sheep.
Evaluation of photoreceptor transduction efficacy of capsid-modified AAV vectors following intravitreal and subretinal delivery in sheep.
复制标题
评估衣壳修饰的 AAV 载体在绵羊玻璃体内和视网膜下递送后的光感受器转导功效。
作者：
M. Ross;A. Obolensky;E. Averbukh;R. Ezra‐Elia;E. Yamin;H. Honig;H. Dvir;A. Rosov;W. Hauswirth;E. Gootwine;E. Banin;Ron Ofri
基于视网膜下递送腺相关病毒（AAV）载体的基因增强疗法被证明在治疗几种遗传性视网膜变性中是高效的。然而，由于视网膜下注射造成的潜在并发症和缺点，寻找将所需的遗传插入物递送到视网膜的替代方法具有很大的推动力。一种这样的方法是载体的玻璃体内递送。我们的目的是评估两种对细胞降解不太敏感的衣壳修饰的载体AAV 8（双Y-F）和AAV 2（quadY-F+T-V）以及第三种嵌合载体AAV[max]在绵羊玻璃体内注射后对感光细胞增殖的功效。我们进一步测试了在玻璃体内注射之前，使用未修饰的载体使内界膜（ILM）病毒结合位点饱和是否会增强光感受器转导的功效。只有AAV[max]在玻璃体内注射后导致中度光感受器转导。玻璃体内注射两种其他载体没有导致光感受器转导，也没有在玻璃体内注射前的ILM的饱和。另一方面，在阳性对照眼中视网膜下注射后，两种载体有效地转导感光细胞。先前在小鼠和犬模型中使用相同载体的试验分别导致了玻璃体内递送后光感受器的稳健和中等转导功效，证明了在评估视网膜基因治疗的新策略时利用尽可能多的动物模型的重要性。玻璃体内注射的AAV[max]的成功光感受器转导使其成为玻璃体内递送的潜在候选者，但需要进一步试验来确定转导功效是否足以获得临床结果。
Gene augmentation therapy based on subretinal delivery of Adeno-Associated Viral (AAV) vectors is proving to be highly efficient in treating several inherited retinal degenerations. However, due to potential complications and drawbacks posed by subretinal injections, there is a great impetus to find alternative methods of delivering the desired genetic inserts to the retina. One such method is an intravitreal delivery of the vector. Our aim was to evaluate the efficacy of two capsid modified vectors that are less susceptible to cellular degradation, AAV8 (doubleY-F) and AAV2 (quadY-F+T-V), as well as a third, chimeric vector AAV[max], to transduce photoreceptor cells following intravitreal injection in sheep. We further tested whether saturation of inner limiting membrane (ILM) viral binding sites using a non-modified vector, prior to the intravitreal injection, would enhance the efficacy of photoreceptor transduction. Only AAV[max] resulted in moderate photoreceptor transduction following intravitreal injection. Intravitreal injection of the two other vectors did not result in photoreceptor transduction, nor did the saturation of the ILM prior to the intravitreal injection. On the other hand, two of the vectors efficiently transduced photoreceptor cells following subretinal injection in positive control eyes. Previous trials with the same vectors in both murine and canine models resulted in robust and moderate transduction efficacy, respectively, of photoreceptors following intravitreal delivery, demonstrating the importance of utilizing as many animal models as possible when evaluating new strategies for retinal gene therapy. The successful photoreceptor transduction of AAV[max] injected intravitreally makes it a potential candidate for intravitreal delivery, but further trials are warranted to determine whether the transduction efficacy is sufficient for a clinical outcome.
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影响因子：
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作者：
Jacobson, Samuel G.;Cideciyan, Artur V.;Ratnakaram, Ramakrishna;Heon, Elise;Schwartz, Sharon B.;Roman, Alejandro J.;Peden, Marc C.;Aleman, Tomas S.;Boye, Sanford L.;Sumaroka, Alexander;Conlon, Thomas J.;Calcedo, Roberto;Pang, Ji-Jing;Erger, Kirsten E.;Olivares, Melani B.;Mullins, Cristina L.;Swider, Malgorzata;Kaushal, Shalesh;Feuer, William J.;Iannaccone, Alessandro;Fishman, Gerald A.;Stone, Edwin M.;Byrne, Barry J.;Hauswirth, William W.
通讯作者：
Hauswirth, William W.
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：
4.2
作者：
Ye, Guo-Jie;Budzynski, Ewa;Chulay, Jeffrey D.
通讯作者：
Chulay, Jeffrey D.
影响因子：
12.4
作者：
Petrs-Silva, Hilda;Dinculescu, Astra;Hauswirth, William W.
通讯作者：
Hauswirth, William W.