Synthetic Monopartite Peptide That Enables the Nuclear Import of Genes Delivered by the Neurotensin-Polyplex Vector,Molecular Pharmaceutics

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Synthetic Monopartite Peptide That Enables the Nuclear Import of Genes Delivered by the Neurotensin-Polyplex Vector
Molecular Pharmaceutics ( IF 4.5 ) Pub Date : 2020-10-30 , DOI: 10.1021/acs.molpharmaceut.0c00755
Francisco E Lopez-Salas ₁ , Rasajna Nadella ₂ , Minerva Maldonado-Berny ₃ , Maria L Escobedo-Sanchez ₃ , Rosana Fiorentino-Pérez ₃ , Bismark Gatica-García ₃ , Manuel A Fernandez-Parrilla ₃ , Moreno Mario Gil ₄ , David Reyes-Corona ₃ , Ubaldo García ₃ , Carlos E Orozco-Barrios ₅ , Maria E Gutierrez-Castillo ₆ , Daniel Martinez-Fong _{1,

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Affiliation

Neurotensin (NTS)-polyplex is a multicomponent nonviral vector that enables gene delivery via internalization of the neurotensin type 1 receptor (NTSR1) to dopaminergic neurons and cancer cells. An approach to improving its therapeutic safety is replacing the viral karyophilic component (peptide KPSV40; MAPTKRKGSCPGAAPNKPK), which performs the nuclear import activity, by a shorter synthetic peptide (KPRa; KMAPKKRK). We explored this issue and the mechanism of plasmid DNA translocation through the expression of the green fluorescent protein or red fluorescent protein fused with KPRa and internalization assays and whole-cell patch-clamp configuration experiments in a single cell together with importin α/β pathway blockers. We showed that KPRa electrostatically bound to plasmid DNA increased the transgene expression compared with KPSV40 and enabled nuclear translocation of KPRa-fused red fluorescent proteins and plasmid DNA. Such translocation was blocked with ivermectin or mifepristone, suggesting importin α/β pathway mediation. KPRa also enabled NTS-polyplex-mediated expression of reporter or physiological genes such as human mesencephalic-derived neurotrophic factor (hMANF) in dopaminergic neurons in vivo. KPRa is a synthetic monopartite peptide that showed nuclear import activity in NTS-polyplex vector-mediated gene delivery. KPRa could also improve the transfection of other nonviral vectors used in gene therapy.

中文翻译：

使神经降压素复合载体提供的基因的核导入成为可能的合成单部分肽

神经降压素 (NTS)-polyplex 是一种多组分非病毒载体，可通过将神经降压素 1 型受体 (NTSR1) 内化到多巴胺能神经元和癌细胞来实现基因传递。提高其治疗安全性的一种方法是用较短的合成肽（KPRa；KMAPKKRK）替代具有核输入活性的病毒亲核成分（肽 KPSV40；MAPTKRKGSCPGAAPNKPK）。我们通过表达与 KPRa 融合的绿色荧光蛋白或红色荧光蛋白以及内化分析和全细胞膜片钳配置实验以及导入蛋白 α/β 通路阻滞剂在单细胞中探讨了这个问题和质粒 DNA 易位的机制. 我们表明，与 KPSV40 相比，KPRa 与质粒 DNA 静电结合增加了转基因表达，并使 KPRa 融合红色荧光蛋白和质粒 DNA 的核易位成为可能。这种易位被伊维菌素或米非司酮阻断，表明导入α/β途径介导。KPRa 还启用了 NTS-polyplex 介导的报告基因或生理基因的表达，例如体内多巴胺能神经元中的人中脑源性神经营养因子 (hMANF)。KPRa 是一种合成的单部分肽，在 NTS 复合载体介导的基因传递中显示出核输入活性。KPRa 还可以改善用于基因治疗的其他非病毒载体的转染。表明输入α/β途径介导。KPRa 还启用了 NTS-polyplex 介导的报告基因或生理基因的表达，例如体内多巴胺能神经元中的人中脑源性神经营养因子 (hMANF)。KPRa 是一种合成的单部分肽，在 NTS 复合载体介导的基因传递中显示出核输入活性。KPRa 还可以改善用于基因治疗的其他非病毒载体的转染。表明输入α/β途径介导。KPRa 还启用了 NTS-polyplex 介导的报告基因或生理基因的表达，例如体内多巴胺能神经元中的人中脑源性神经营养因子 (hMANF)。KPRa 是一种合成的单部分肽，在 NTS 复合载体介导的基因传递中显示出核输入活性。KPRa 还可以改善用于基因治疗的其他非病毒载体的转染。

更新日期：2020-12-07

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