PEGylated enhanced cell penetrating peptide nanoparticles for lung gene therapy.
PEGylated enhanced cell penetrating peptide nanoparticles for lung gene therapy.
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DOI：
10.1016/j.jconrel.2018.07.001
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发表时间：
2018-09-10
期刊：
影响因子：
--
通讯作者：
Dixon JE
中科院分区：
文献类型：
--
作者：
Osman G;Rodriguez J;Chan SY;Chisholm J;Duncan G;Kim N;Tatler AL;Shakesheff KM;Hanes J;Suk JS;Dixon JE
对于囊性纤维化等单基因疾病的基因治疗，肺仍然是一个有吸引力的靶点。尽管有超过27项临床试验，但仍有极少数基因治疗载体显示肺功能有任何改善；这突显了开发具有更高基因转移效力和所需物理化学特性的配方以实现有效治疗的必要性。在这里，我们介绍了一种新型的基于细胞穿透肽(CPP)的非病毒载体，它利用糖胺聚糖(GAG)结合的增强型转导(GET)进行高效的基因转移。通过静电作用将多肽与DNA直接偶联，形成纳米粒子。为了使GET多肽能够在体内高效传递，我们设计了多肽的聚乙二醇化版本，并采用了一种策略来形成具有不同密度的聚乙二醇包被物的DNA纳米粒。我们能够确定候选制剂(聚乙二醇化率≥为40%)，该制剂能够屏蔽颗粒表面的正电荷，保持在支气管肺泡灌洗液中的胶体稳定性，并在体外培养的人支气管上皮细胞株和精密切割肺切片中保持基因转移活性。利用多粒子跟踪(MPT)技术，我们证明了聚乙二醇GET复合体能够在体外通过粘液网状结构并在患者的CF痰样本中快速扩散。在活体小鼠肺模型中测试时，与非聚乙二醇化复合体相比，聚乙二醇化颗粒表现出更好的生物分布、改善的安全性和报告荧光素酶质粒的高效基因转移。此外，与聚乙烯亚胺(PEI)相比，基因表达显著增强，PEI是一种已在临床前环境中广泛测试的非病毒基因载体。这项工作描述了一种创新的方法，将用于增强转染的新型GET多肽与可调节的聚乙二醇包被物相结合，用于有效的肺基因治疗。
The lung remains an attractive target for the gene therapy of monogenetic diseases such as cystic fibrosis (CF). Despite over 27 clinical trials, there are still very few gene therapy vectors that have shown any improvement in lung function; highlighting the need to develop formulations with improved gene transfer potency and the desirable physiochemical characteristics for efficacious therapy. Herein, we introduce a novel cell penetrating peptide (CPP)-based non-viral vector that utilises glycosaminoglycan (GAG)-binding enhanced transduction (GET) for highly efficient gene transfer. GET peptides couple directly with DNA through electrostatic interactions to form nanoparticles (NPs). In order to adapt the GET peptide for efficient in vivo delivery, we engineered PEGylated versions of the peptide and employed a strategy to form DNA NPs with different densities of PEG coatings. We were able to identify candidate formulations (PEGylation rates ≥ 40%) that shielded the positively charged surface of particles, maintained colloidal stability in bronchoalveolar lavage fluid (BALF) and retained gene transfer activity in human bronchial epithelial cell lines and precision cut lung slices (PCLS) in vitro. Using multiple particle tracking (MPT) technology, we demonstrated that PEG-GET complexes were able to navigate the mucus mesh and diffuse rapidly through patient CF sputum samples ex vivo. When tested in mouse lung models in vivo, PEGylated particles demonstrated superior biodistribution, improved safety profiles and efficient gene transfer of a reporter luciferase plasmid compared to non-PEGylated complexes. Furthermore, gene expression was significantly enhanced in comparison to polyethylenimine (PEI), a non-viral gene carrier that has been widely tested in pre-clinical settings. This work describes an innovative approach that combines novel GET peptides for enhanced transfection with a tuneable PEG coating for efficacious lung gene therapy.
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DOI：
10.1056/nejmra0910061
发表时间：
2010-12-02
期刊：
The New England journal of medicine
影响因子：
--
作者：
Fahy JV;Dickey BF
通讯作者：
Dickey BF
影响因子：
10.8
作者：
Kleemann, E;Neu, M;Kissel, T
通讯作者：
Kissel, T
DOI：
10.1016/j.jconrel.2016.05.031
发表时间：
2016-10-28
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Kim N;Duncan GA;Hanes J;Suk JS
通讯作者：
Suk JS
影响因子：
4.9
作者：
Lee, Jue-Yeon;Choo, Jung-Eun;Park, Yoon-Jeong
通讯作者：
Park, Yoon-Jeong
影响因子：
12.7
作者：
Kajon, AE;Gigliotti, AP;Harrod, KS
通讯作者：
Harrod, KS