Ferla Rita, Dell'Aquila Fabio, Doria Monica, Ferraiuolo Maria, Noto Alessia, Grazioli Fabiana, Ammendola Virginia, Testa Francesco, Melillo Paolo, Iodice Carolina, Risca Giulia, Tedesco Novella, le Brun Pierre Romain, Surace Enrico Maria, Simonelli Francesca, Galimberti Stefania, Valsecchi Maria Grazia, Marteau Jean-Brice, Veron Philippe, Colloca Stefano, Auricchio Alberto
Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078 Pozzuoli, Italy.
AAVantgarde BIO Srl, 20123 Milan, Italy.
Mol Ther Methods Clin Dev. 2023 Feb 11;28:396-411. doi: 10.1016/j.omtm.2023.02.002. eCollection 2023 Mar 9.
Gene therapy of Usher syndrome type 1B (USH1B) due to mutations in the large () gene is limited by the packaging capacity of adeno-associated viral (AAV) vectors. To overcome this, we have previously developed dual AAV8 vectors which encode human (dual AAV8.). Here we show that subretinal administration of 1.37E+9 to 1.37E+10 genome copies of a good-manufacturing-practice-like lot of dual AAV8. improves the retinal defects of a mouse model of USH1B. The same lot was used in non-human primates at doses 1.6× and 4.3× the highest dose proposed for the clinical trial which was based on mouse efficacy data. Long-lasting alterations in retinal function and morphology were observed following subretinal administration of dual AAV8. at the high dose. These findings were modest and improved over time in the low-dose group, as also observed in other studies involving the use of AAV8 in non-human primates and humans. Biodistribution and shedding studies confirmed the presence of vector DNA mainly in the visual pathway. Accordingly, we detected human MYO7A mRNA expression predominantly in the retina. Overall, these studies pave the way for the clinical translation of subretinal administration of dual AAV vectors in USH1B subjects.
由于大的（）基因突变导致的1B型Usher综合征（USH1B）的基因治疗受到腺相关病毒（AAV）载体包装能力的限制。为克服这一问题，我们之前开发了编码人（双AAV8.）的双AAV8载体。在此我们表明，向视网膜下注射1.37E+9至1.37E+10基因组拷贝的类似药品生产质量管理规范批次的双AAV8.可改善USH1B小鼠模型的视网膜缺陷。基于小鼠疗效数据，将同一批次用于非人灵长类动物，剂量为临床试验提议的最高剂量的1.6倍和4.3倍。视网膜下注射高剂量双AAV8.后，观察到视网膜功能和形态出现持久改变。这些发现并不显著，且低剂量组随时间推移有所改善，在其他涉及在非人灵长类动物和人类中使用AAV8的研究中也观察到了这一点。生物分布和脱落研究证实载体DNA主要存在于视觉通路中。因此，我们主要在视网膜中检测到了人MYO7A mRNA表达。总体而言，这些研究为在USH1B患者中视网膜下注射双AAV载体的临床转化铺平了道路。
