Peptide-Functionalized Dendrimer Nanocarriers for Targeted Microdystrophin Gene Delivery.
Peptide-Functionalized Dendrimer Nanocarriers for Targeted Microdystrophin Gene Delivery.
复制标题
靶向递送微营养不良蛋白基因的多肽功能化树状聚合物纳米载体。
DOI：
10.3390/pharmaceutics13122159
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发表时间：
2021-12-15
期刊：
影响因子：
5.4
通讯作者：
Deo SK
中科院分区：
文献类型：
--
作者：
Hersh J;Condor Capcha JM;Iansen Irion C;Lambert G;Noguera M;Singh M;Kaur A;Dikici E;Jiménez JJ;Shehadeh LA;Daunert S;Deo SK
基因治疗是确定的先天性疾病的一个很好的替代方案；然而，体内基因传递有许多限制，包括靶向细胞摄取、细胞内转运和通过核膜的运输。在这里，我们开发了一种改进的G5聚酰胺胺(G5 PAMAM)树枝状大分子-DNA复合体，它将允许细胞特异性靶向骨骼肌细胞，并通过细胞内机制和核膜运输DNA。用骨骼肌靶向多肽(SMTP)、DLC8结合多肽(DBP)和核定位信号转导多肽(NLS)修饰G5PAMAM纳米载体，并与含有GFP标记的微肌营养不良蛋白(µDys)基因的质粒DNA进行复合。µDys的交付一直被认为是治疗衰弱性Duchenne肌营养不良症(DMD)患者的一种方式。制备了不同电荷比的纳米载体-多肽-DNA复合体，并对其稳定性、大小、表面电荷和细胞毒性进行了表征。利用优化的纳米载体复合体，通过荧光和Western blotting分别检测GFP和µDys蛋白的表达，以确定体外的转染率。通过向Duchenne模型小鼠注射最佳的纳米载体复合制剂mdx4Cv来检测体内蛋白质的表达。最终，这些纳米载体复合体将允许将微肌营养不良蛋白基因定向传递到骨骼肌细胞，并改善Duchenne肌营养不良患者的肌肉功能。
Gene therapy is a good alternative for determined congenital disorders; however, there are numerous limitations for gene delivery in vivo including targeted cellular uptake, intracellular trafficking, and transport through the nuclear membrane. Here, a modified G5 polyamidoamine (G5 PAMAM) dendrimer–DNA complex was developed, which will allow cell-specific targeting to skeletal muscle cells and transport the DNA through the intracellular machinery and the nuclear membrane. The G5 PAMAM nanocarrier was modified with a skeletal muscle-targeting peptide (SMTP), a DLC8-binding peptide (DBP) for intracellular transport, and a nuclear localization signaling peptide (NLS) for nuclear uptake, and polyplexed with plasmid DNA containing the GFP-tagged microdystrophin (µDys) gene. The delivery of µDys has been considered as a therapeutic modality for patients suffering from a debilitating Duchenne muscular dystrophy (DMD) disorder. The nanocarrier–peptide–DNA polyplexes were prepared with different charge ratios and characterized for stability, size, surface charge, and cytotoxicity. Using the optimized nanocarrier polyplexes, the transfection efficiency in vitro was determined by demonstrating the expression of the GFP and the µDys protein using fluorescence and Western blotting studies, respectively. Protein expression in vivo was determined by injecting an optimal nanocarrier polyplex formulation to Duchenne model mice, mdx4Cv. Ultimately, these nanocarrier polyplexes will allow targeted delivery of the microdystrophin gene to skeletal muscle cells and result in improved muscle function in Duchenne muscular dystrophy patients.
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DOI：
10.1111/febs.12072
发表时间：
2013-02
期刊：
The FEBS journal
影响因子：
--
作者：
Jackson MF;Hoversten KE;Powers JM;Trobridge GD;Rodgers BD
通讯作者：
Rodgers BD
DOI：
10.1073/pnas.86.4.1292
发表时间：
1989-02-01
影响因子：
11.1
作者：
CHAPMAN, VM;MILLER, DR;CASKEY, CT
通讯作者：
CASKEY, CT
影响因子：
3.6
作者：
Condor Capcha JM;Lambert G;Dykxhoorn DM;Salerno AG;Hare JM;Whitt MA;Pahwa S;Jayaweera DT;Shehadeh LA
通讯作者：
Shehadeh LA
影响因子：
11.2
作者：
Daftarian, Pirouz;Kaifer, Angel E.;Serafini, Paolo
通讯作者：
Serafini, Paolo
影响因子：
15.6
作者：
Fox, Laura J.;Richardson, Robert M.;Briscoe, Wuge H.
通讯作者：
Briscoe, Wuge H.