Gene delivery to mitotic and postmitotic photoreceptors via compacted DNA nanoparticles results in improved phenotype in a mouse model of retinitis pigmentosa
Gene delivery to mitotic and postmitotic photoreceptors via compacted DNA nanoparticles results in improved phenotype in a mouse model of retinitis pigmentosa
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DOI：
10.1096/fj.09-139147
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发表时间：
2010-04-01
期刊：
影响因子：
4.8
通讯作者：
Naash, Muna I.
中科院分区：
文献类型：
--
作者：
Cai, Xue;Conley, Shannon M.;Naash, Muna I.
本研究的目的是测试经修饰的DNA纳米颗粒介导的基因递送到视网膜色素变性的幼年小鼠模型的视网膜下腔中的治疗效率和安全性。将含有小鼠视蛋白启动子和野生型小鼠Rds基因的纳米颗粒在出生后第5天和第22天（P）视网膜下注射到在视网膜变性慢（rds（+/-））基因中携带单倍不足突变的小鼠中。对照组小鼠要么注射生理盐水，要么注射携带Rds基因的未压缩裸质粒DNA，要么保持不治疗。对于P5注射，Rds mRNA水平在注射后第2至7天（PI-2至PI-7）达到峰值，对于P5和P22注射，Rds mRNA水平稳定在比未注射对照高2倍的水平，并且在检查的最后时间点（PI-120）保持升高。在P5和P22注射后，与对照组相比，视杆功能（通过视网膜电图测量）显示出适度但统计学显著的改善。注射纳米颗粒的眼睛中的视锥细胞功能在两个注射年龄段都达到了野生型水平，表明完全预防了视锥细胞变性。PI-120的超微结构检查显示P5纳米颗粒注射眼的外节结构有显著改善，而P22注射有适度的结构改善。在PI-2或PI-30时，在P5或P22纳米颗粒给药的眼中没有巨噬细胞活化或IL-6或TNF-α mRNA诱导的证据。因此，固定的DNA纳米颗粒可以有效和安全地驱动有丝分裂和有丝分裂后光感受器中的基因表达，并延缓该模型中的变性。这些发现，使用临床相关的治疗范例，说明了应用基于纳米颗粒的基因替代疗法治疗人类视网膜变性的潜力。蔡某，Conley，S. M.，纳什，Z.，Fliesler，S. J.，库珀，M. J.，Naash，M. I.通过致密的DNA纳米颗粒将基因传递到有丝分裂和有丝分裂后的光感受器导致视网膜色素变性小鼠模型的表型改善。FASEB J.24，1178-1191（2010）。www.fasebj.org
The purpose of the present study was to test the therapeutic efficiency and safety of compacted-DNA nanoparticle-mediated gene delivery into the sub-retinal space of a juvenile mouse model of retinitis pigmentosa. Nanoparticles containing the mouse opsin promoter and wild-type mouse Rds gene were injected subretinally into mice carrying a haploinsufficiency mutation in the retinal degeneration slow (rds(+/-)) gene at postnatal day (P) 5 and 22. Control mice were either injected with saline, injected with uncompacted naked plasmid DNA carrying the Rds gene, or remained untreated. Rds mRNA levels peaked at postinjection day 2 to 7 (PI-2 to PI-7) for P5 injections, stabilized at levels 2-fold higher than in uninjected controls for both P5 and P22 injections, and remained elevated at the latest time point examined (PI-120). Rod function (measured by electroretinography) showed modest but statistically significant improvement compared with controls after both P5 and P22 injections. Cone function in nanoparticle-injected eyes reached wild-type levels for both ages of injections, indicating full prevention of cone degeneration. Ultrastructural examination at PI-120 revealed significant improvement in outer segment structures in P5 nanoparticle-injected eyes, while P22 injection had a modest structural improvement. There was no evidence of macrophage activation or induction of IL-6 or TNF-alpha mRNA in P5 or P22 nanoparticle-dosed eyes at either PI-2 or PI-30. Thus, compacted-DNA nanoparticles can efficiently and safely drive gene expression in both mitotic and postmitotic photoreceptors and retard degeneration in this model. These findings, using a clinically relevant treatment paradigm, illustrate the potential for application of nanoparticle-based gene replacement therapy for treatment of human retinal degenerations.-Cai, X., Conley, S. M., Nash, Z., Fliesler, S. J., Cooper, M. J., Naash, M. I. Gene delivery to mitotic and postmitotic photoreceptors via compacted DNA nanoparticles results in improved phenotype in a mouse model of retinitis pigmentosa. FASEB J. 24, 1178-1191 (2010). www.fasebj.org