Ho Thucanh T, Maguire Albert M, Aguirre Gustavo D, Surace Enrico M, Anand Vibha, Zeng Yong, Salvetti Anna, Hopwood John J, Haskins Mark E, Bennett Jean
F.M. Kirby Center for Molecular Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia, PA, USA.
J Gene Med. 2002 Nov-Dec;4(6):613-21. doi: 10.1002/jgm.302.
Mucopolysaccharidosis VI (MPS VI), due to recessively inherited 4-sulfatase (4S) deficiency, results in lysosomal storage of dermatan sulfate in numerous tissues. Retinal involvement is limited to the retinal pigment epithelium (RPE). This study aimed to determine whether recombinant adeno-associated virus (AAV)-mediated delivery of 4S would reverse the RPE pathology seen in MPS VI cats.
AAV.f4S, containing the feline 4S cDNA, was delivered unilaterally to eyes of affected cats by subretinal or intravitreal injection. Contralateral eyes received AAV with the green fluorescent protein (GFP) reporter gene as control. At 2-11 months post-injection, the cats were sacrificed and the treatment effects were evaluated histologically.
By ophthalmoscopy and histological analyses, GFP was evident as early as 4 weeks and persisted through the latest time point (11 months). Untreated and AAV.GFP-treated diseased retinas contained massively hypertrophied RPE cells secondary to accumulation of dilated lysosomal inclusions containing dermatan sulfate. MPS VI eyes treated subretinally with AAV.f4S had minimal RPE cell inclusions and, consequently, were not hypertrophied.
AAV-mediated subretinal delivery of f4S provided correction of the disease phenotype in RPE cells of feline MPS VI, supporting the utility of AAV as a vector for the treatment of RPE-specific as well as lysosomal storage diseases.
黏多糖贮积症 VI 型（MPS VI）是由隐性遗传的 4 - 硫酸酯酶（4S）缺乏引起的，导致硫酸皮肤素在许多组织的溶酶体中蓄积。视网膜受累仅限于视网膜色素上皮（RPE）。本研究旨在确定重组腺相关病毒（AAV）介导的 4S 递送是否能逆转 MPS VI 型猫眼中所见的 RPE 病理变化。
将含有猫 4S cDNA 的 AAV.f4S 通过视网膜下或玻璃体内注射单侧递送至患病猫的眼睛。对侧眼睛接受带有绿色荧光蛋白（GFP）报告基因的 AAV 作为对照。在注射后 2 - 11 个月，处死猫并通过组织学评估治疗效果。
通过检眼镜检查和组织学分析，GFP 最早在 4 周时就很明显，并持续到最晚的时间点（11 个月）。未经治疗和接受 AAV.GFP 治疗的患病视网膜含有大量肥大的 RPE 细胞，这是由于含有硫酸皮肤素的扩张溶酶体包涵体蓄积所致。用 AAV.f4S 进行视网膜下治疗的 MPS VI 型眼睛的 RPE 细胞包涵体极少，因此没有肥大。
AAV 介导的视网膜下递送 f4S 纠正了猫 MPS VI 型 RPE 细胞中的疾病表型，支持 AAV 作为治疗 RPE 特异性以及溶酶体贮积病的载体的实用性。
