Formulation and efficacy of ECO/pRHO-ABCA4-SV40 nanoparticles for nonviral gene therapy of Stargardt disease in a mouse model.
Formulation and efficacy of ECO/pRHO-ABCA4-SV40 nanoparticles for nonviral gene therapy of Stargardt disease in a mouse model.
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DOI：
10.1016/j.jconrel.2020.12.010
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发表时间：
2021-02-10
影响因子：
10.8
通讯作者：
Lu, Zheng-Rong
中科院分区：
文献类型：
--
作者：
Sun, Da;Sun, Wenyu;Gao, Song-Qi;Wei, Cheng;Naderi, Amirreza;Schilb, Andrew L.;Scheidt, Josef;Lee, Sangjoon;Kern, Timothy S.;Palczewski, Krzysztof;Lu, Zheng-Rong
对于由大基因突变引起的视网膜疾病，如Stargardt病（STGD），开发基因替代疗法仍然是一个挑战。STGD是由ABCA 4基因突变引起的。在此之前，我们已经开发了一种有效的非病毒基因治疗，使用自组装纳米粒子的多功能pH敏感的氨基脂质ECO和治疗ABCA 4质粒含有视紫红质启动子（pRHO-ABCA 4）。在本研究中，我们用猿猴病毒40增强子（SV 40，pRH 0-ABCA 4-SV 40）修饰ABCA 4质粒以增强基因表达。我们还使用蔗糖或山梨糖醇作为稳定剂制备和评估ECO/pDNA纳米颗粒的制剂以开发一致和稳定的制剂。结果表明，ECO在蔗糖存在下与pRH 0-ABCA 4-SV 40形成稳定的纳米颗粒，但不与山梨醇形成。在含有CMV启动子的质粒pCMV-ABCA 4-SV 40中引入SV 40增强子后，体外转染效率显著提高。蔗糖对转染效率无影响，山梨醇对转染效率有影响。使用ECO/pRHO-ABCA 4和ECO/pRHO-ABCA 4-SV 40纳米颗粒对Abca 4 −/−小鼠进行视网膜下基因治疗，分别诱导A2 E蓄积减少36%和29%。因此，基于ECO/pABCA 4的纳米颗粒有希望用于Stargardt病的非病毒基因治疗，并且可以扩展用于具有突变的大基因的各种视觉营养不良的应用。
It is still a challenge to develop gene replacement therapy for retinal disorders caused by mutations in large genes, such as Stargardt disease (STGD). STGD is caused by mutations in ABCA4 gene. Previously, we have developed an effective non-viral gene therapy using self-assembled nanoparticles of a multifunctional pH-sensitive amino lipid ECO and a therapeutic ABCA4 plasmid containing rhodopsin promoter (pRHO–ABCA4). In this study, we modified the ABCA4 plasmid with simian virus 40 enhancer (SV40, pRHO–ABCA4–SV40) for enhanced gene expression. We also prepared and assessed the formulations of ECO/pDNA nanoparticles using sucrose or sorbitol as a stablilizer to develop consistent and stable formulations. Results demonstrated that ECO formed stable nanoparticles with pRHO–ABCA4–SV40 in the presence of sucrose, but not with sorbitol. The transfection efficiency in vitro increased significantly after introduction of SV40 enhancer for plasmid pCMV-ABCA4–SV40 with a CMV promoter. Sucrose didn’t affect the transfection efficiency, while sorbitol resulted in a fluctuation of the in vitro transfection efficiency. Subretinal gene therapy in Abca4−/− mice using ECO/pRHO–ABCA4 and ECO/pRHO–ABCA4–SV40 nanoparticles induced 36% and 29% reduction in A2E accumulation respectively. Therefore, the ECO/pABCA4 based nanoparticles are promising for non-viral gene therapy for Stargardt disease and can be expended for applications in a variety of visual dystrophies with mutated large genes.
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影响因子：
4
作者：
Ayat, Nadia R.;Sun, Zhanhu;Lu, Zheng-Rong
通讯作者：
Lu, Zheng-Rong
影响因子：
10.8
作者：
Mashal, Mohamed;Attia, Noha;Luis Pedraz, Jose
通讯作者：
Luis Pedraz, Jose
影响因子：
3.7
作者：
Piedmonte, Deirdre Murphy;Summers, Christie;Ratnaswamy, Gayathri
通讯作者：
Ratnaswamy, Gayathri
影响因子：
14.9
作者：
DEVILLIERS, J;OLSON, L;SCHAFFNER, W
通讯作者：
SCHAFFNER, W
影响因子：
3.4
作者：
Bonaccorso, A.;Musumeci, T.;Puglisi, G.
通讯作者：
Puglisi, G.