AAV-Mediated Gene Replacement, Either Alone or in Combination with Physical and Pharmacological Agents, Results in Partial and Transient Protection from Photoreceptor Degeneration Associated with βPDE Deficiency
AAV-Mediated Gene Replacement, Either Alone or in Combination with Physical and Pharmacological Agents, Results in Partial and Transient Protection from Photoreceptor Degeneration Associated with βPDE Deficiency
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DOI：
10.1167/iovs.10-6269
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发表时间：
2011-07-01
影响因子：
4.4
通讯作者：
Auricchio, Alberto
中科院分区：
文献类型：
--
作者：
Allocca, Mariacarmela;Manfredi, Anna;Auricchio, Alberto
目的。PDE6B基因突变导致隐性严重视网膜色素变性（RP）。PDE6B编码杆状特异性磷酸二酯酶（PDE）的β亚基，当PDE缺失时，会导致细胞内Ca2+的毒性水平和光感受器细胞死亡。Ca2+阻滞剂，如尼伐地平，以及光限制，减缓β - PDE缺乏动物模型的光感受器变性。该研究的目的是评估aav2 /5或aav2 /8介导的基因替代联合尼伐地平和/或限光对携带纯合子pde6b突变的rd10小鼠的疗效。编码β - PDE或EGFP的AAV载体在出生后(P)2给予视网膜下注射。尼伐地平于P7至P28给予。对于光照限制，怀孕的rd10小鼠被关在黑暗环境中，直到它们的幼崽28天大。所有功能和组织学分析均在p35进行。在视网膜下给药编码EGFP的AAV载体后，观察到明显的形态光感受器保护。在使用AAV2/8或-2/5编码β - PDE后，这种保护进一步增强，但与显著的功能改善无关。与aav2 /5介导的递送相比，AAV2/8-介导的递送对光感受器的保护作用更高，并且通过额外的药物治疗和/或光限制没有显著增强。p35后保护作用消失。总之，应该探索比本研究中使用的更有效的基因转移工具，以及更好地了解疾病的发病机制，以增加基因替代的效果，并设计基于基因的策略，阻断β - PDE缺乏激活的凋亡途径。（Invest Ophthalmol Vis Sci. 2011;52:5713-5719） DOI:10.1167/iovs.10-6269
PURPOSE. Mutations in the PDE6B gene cause recessive, severe retinitis pigmentosa (RP). PDE6B encodes the beta subunit of the rod-specific phosphodiesterase (beta PDE), which, when absent, results in toxic levels of intracellular Ca2+ and photoreceptor cell death. Ca2+ blockers, such as nilvadipine, as well as light restriction, slow photoreceptor degeneration in animal models of beta PDE deficiencies. The goal of the study was to evaluate the efficacy of AAV2/5-or AAV2/8-mediated gene replacement in combination with nilvadipine and/or with light restriction in the rd10 mouse bearing homozygous pde6b mutations.METHODS. AAV vectors encoding either beta PDE or EGFP were subretinally administered at postnatal day (P)2. Nilvadipine was administered from P7 to P28. For light restriction, pregnant rd10 mice were kept in a dark environment until their pups were 28 days old. All functional and histologic analyses were performed at P35.RESULTS. Significant morphologic photoreceptor protection was observed after subretinal administration of AAV vectors encoding EGFP. This protection further increased after administration of AAV2/8 or -2/5 encoding for beta PDE and was not associated with significant functional improvement. Photoreceptor protection was higher after AAV2/8- than after AAV2/5-mediated delivery and was not significantly augmented by additional drug therapy and/or light restriction. The protective effect was lost after P35.CONCLUSIONS. In conclusion, more efficient gene transfer tools than those used in this study, as well as a better understanding of the disease pathogenesis, should be explored to increase the effect of gene replacement and to design gene-based strategies that block the apoptotic pathways activated by beta PDE deficiency. (Invest Ophthalmol Vis Sci. 2011;52:5713-5719) DOI:10.1167/iovs.10-6269