Effects of adeno-associated virus-vectored ciliary neurotrophic factor on retinal structure and function in mice with a P216L rds/peripherin mutation
Effects of adeno-associated virus-vectored ciliary neurotrophic factor on retinal structure and function in mice with a P216L rds/peripherin mutation
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DOI：
10.1006/exer.2002.1176
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发表时间：
2002-06-01
影响因子：
3.4
通讯作者：
Lavail, MM
中科院分区：
文献类型：
--
作者：
Bok, D;Yasumura, D;Lavail, MM
以往的研究表明，睫状神经营养因子（CNTF）的急性给药能够延长遭受光毒性损伤或表达基因突变的视网膜光感受器细胞的存活时间。腺病毒载体介导的CNTF也有效果，但对于所有这些治疗方法，效果都是暂时的。另一方面，腺相关病毒载体介导的小基因在长期存活方面具有相当大的潜力。作者试图通过使用重组腺相关病毒（rAAV）来递送编码分泌型CNTF的小基因，为表达rds基因显性负性点突变的小鼠光感受器提供基于CNTF的长期保护。在巨细胞病毒（CMV）或鸡β - 肌动蛋白（CBA）启动子控制下的分泌型CNTF，在将rAAV载体单次注射到视网膜下腔后，为光感受器提供了长期的全视网膜挽救作用。当载体被放置在玻璃体腔时，挽救效果要差得多且重复性较差。然而，出现了似乎与剂量相关的意外副作用。一个副作用是视杆光感受器细胞核表型的改变，其特征是在视网膜下注射后常染色质增加以及核大小增加，但在玻璃体内注射时没有这种情况。当使用推测更强的CBA启动子时，这些核变化是全视网膜性的，而当使用CMV启动子时则不是全视网膜性的，在后一种情况下，注射部位的核变化更为明显。因此，CNTF对视网膜细胞的慢性过度刺激可能会上调光感受器中的基因表达。根据目前对CNTF视网膜细胞靶点的了解，这种效应可能是间接的，可能并不代表CNTF对光感受器的直接刺激。第二个副作用是，与对侧未注射的视网膜相比，注射并挽救的视网膜在暗视a波和b波振幅以及明视b波振幅方面出现了矛盾性的降低，尽管这些视网膜比未处理的视网膜具有更多的光感受器。这些视网膜电图（ERG）振幅降低的原因可能与基因表达的变化有关。在考虑将CNTF用于改善遗传性视网膜变性的人体临床试验之前，应该确定这些副作用的机制以及CNTF的适当给药剂量。（C）2002爱思唯尔科学有限公司
Past studies have shown that acute administration of ciliary neurotrophic factor (CNTF) can prolong the survival of retinal photoreceptor cells that have undergone phototoxic injury or that express gene mutations. Adenovirus-vectored CNTF has also been effective but for all of these treatments, the effect has been transient. On the other hand, adeno-associated virus-vectored minigenes offer considerable promise for long-term survival. The authors sought to provide long-term. CNTF-based protection of mouse photorcceptors expressing a dominant-negative point mutation in the rds gene by using recombinant adeno-associated virus (rAAV) to deliver minigenes that code for a secreted form of CNTF.Secreted CNTF, under control of a cytomegalovirus (CMV) or chick beta actin (CBA) promoter provided long-term, panretinal rescue of photoreceptors following single injections of rAAV vectors into the subretinal compartment. Rescue was much less effective and less reproducible when the vectors were placed in the vitreous compartment. However, there were unexpected side effects that appeared to be dose-related. One side effect was a change in rod photoreceptor nucleus phenotype, featuring an increase in euchromatin and an increase in nuclear size following subretinal injections but not intravitreal injections. These nuclear changes were panretinal when the putatively stronger CBA promoter was used but not panretinal when the CMV promoter was used. In the latter case, the nuclear changes were much more pronounced at the site of injection. Thus, chronic hyperstimulation of retinal cells with CNTF may up-regulate gene expression in photoreceptors. Based on current knowledge of retinal cell targets for CNTF, this effect may be indirect and may not represent direct stimulation of photoreceptors by CNTF.A second side effect was a paradoxical decrease in scotopic a- and b-wave amplitudes and a decrease in photopic b-wave amplitudes in the injected, rescued retina when compared to its contralateral, uninjected counterpart, in spite of the fact that these retinas had more photoreceptors than their untreated mates. The basis for these decreased ERG amplitudes may be related to changes in gene expression. The mechanisms for these side effects and proper doses of CNTF administration should be determined before human clinical trials are considered for the amelioration of inherited retinal degenerations with CNTF. (C) 2002 Elsevier Science Ltd.