Department of Otolaryngology-Head and Neck Surgery, University of California, San Francisco, CA.
Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, FL 32610.
Proc Natl Acad Sci U S A. 2019 Mar 5;116(10):4496-4501. doi: 10.1073/pnas.1817537116. Epub 2019 Feb 19.
Autosomal recessive genetic forms (DFNB) account for most cases of profound congenital deafness. Adeno-associated virus (AAV)-based gene therapy is a promising therapeutic option, but is limited by a potentially short therapeutic window and the constrained packaging capacity of the vector. We focus here on the otoferlin gene underlying DFNB9, one of the most frequent genetic forms of congenital deafness. We adopted a dual AAV approach using two different recombinant vectors, one containing the 5' and the other the 3' portions of otoferlin cDNA, which exceed the packaging capacity of the AAV when combined. A single delivery of the vector pair into the mature cochlea of mutant mice reconstituted the otoferlin cDNA coding sequence through recombination of the 5' and 3' cDNAs, leading to the durable restoration of otoferlin expression in transduced cells and a reversal of the deafness phenotype, raising hopes for future gene therapy trials in DFNB9 patients.
常染色体隐性遗传形式（DFNB）占大多数严重先天性耳聋病例。腺相关病毒（AAV）为基础的基因治疗是一种很有前途的治疗选择，但受到潜在的治疗窗口短和载体的有限包装能力的限制。我们在这里关注的是 otoferlin 基因，该基因是先天性耳聋最常见的遗传形式之一。我们采用了双 AAV 方法，使用两种不同的重组载体，一种载体包含 otoferlin cDNA 的 5' 端，另一种载体包含 3' 端，当两者结合时，超过了 AAV 的包装能力。将载体对单次递送到突变型小鼠的成熟耳蜗中，通过 5' 和 3' cDNA 的重组，重新构建 otoferlin cDNA 编码序列，导致转导细胞中 otoferlin 表达的持久恢复，并逆转耳聋表型，为未来在 DFNB9 患者中进行基因治疗试验带来了希望。
