Poly(Propyleneimine) Transposon-Nanocarriers for Targeted Delivery of Therapeutic DNAs to Tumor Cells

用于将治疗性 DNA 靶向递送至肿瘤细胞的聚（丙烯亚胺）转座子纳米载体

• 批准号: 419035191
• 负责人: Privatdozent Dr. Dietmar Appelhans, Ph.D.
• 金额: --
• 依托单位: Leibniz-Institut für Polymerforschung Dresden (IPF)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2023-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/419035191?language=en
• 关键词: Poly; Propyleneimine; Transposon; Nanocarriers; Targeted

The delivery of therapeutic nucleic acids, such as DNAs encoding suicide genes or shRNAs using polycationic carrier macromolecules, such as perfectly-branched poly(propyleneimine) dendrimers (PPI), is a promising therapeutic option to treat cancer. However, systemic delivery of carrier-complexed nucleic acids requires huge amounts of DNA which often results in unwanted unspecific off-tissue and off-target effects. This is particularly so since the transient expression of DNA also imply that repeated systemic administration is mandatory for treatment. Yet, the risk of cumulative toxicity is expected to increase. One approach to avoid unwanted off-target effects and to provide stable expression of DNA is the introduction of targeting moieties (ligands, antibodies) that specifically bind to target cells and deliver the therapeutic gene, using the concept of targeted delivery.In our project we plan to investigate a novel targeted transposon-minicircle DNA-delivery approach using functionalized PPIs enabling selective uptake and endosomal release in target cells displaying a surface expression of the cognate cellular receptor. More specifically, functionalized PPIs, coupled to tumor-selective targeting moieties such as single chain fragment variables (scFv) and peptide ligands, will be employed to selectively target glioma and prostate carcinoma tumor cells. The stable expression of the therapeutic genes is provided by co-delivery of a sleeping beauty transposase. In this proof-of-concept study the selective PPI-based transposon-nanocarrier platform will be investigated in vitro using gene transfer of reporter genes (EGFP, Luciferase), a suicide gene (Pseudomonas aeruginosa exotoxin PE38) and DNAs encoding for therapeutic shRNAs. The objective of this project therefore is to establish a biocompatible PPI-based -transposon carriers as a platform for the selective delivery of genes into target cells.

使用聚阳离子载体大分子（例如完全支化的聚（丙烯亚胺）树枝状聚合物（PPI））递送治疗性核酸（例如编码自杀基因的DNA或shRNA）是治疗癌症的有希望的治疗选择。然而，载体复合的核酸的全身递送需要大量的DNA，这通常导致不希望的非特异性组织外和脱靶效应。尤其如此，因为DNA的瞬时表达也意味着重复的全身给药对于治疗是强制性的。然而，累积毒性的风险预计会增加。避免不希望的脱靶效应并提供DNA的稳定表达的一种方法是引入靶向部分（配体，抗体）特异性结合靶细胞并递送治疗基因，在我们的项目中，我们计划研究一种新的靶向转座子-微环DNA，使用官能化PPI的递送方法，能够在显示同源细胞受体的表面表达的靶细胞中选择性摄取和内体释放。更具体地说，与肿瘤选择性靶向部分（例如单链片段可变体（scFv）和肽配体）偶联的功能化PPI将用于选择性靶向神经胶质瘤和前列腺癌肿瘤细胞。通过共递送睡美人转座酶来提供治疗基因的稳定表达。在这项概念验证研究中，将使用报告基因（EGFP，荧光素酶）、自杀基因（铜绿假单胞菌外毒素PE 38）和编码治疗性shRNA的DNA的基因转移，在体外研究选择性PPI为基础的转座子-纳米载体平台。因此，本项目的目的是建立一个生物相容性的PPI为基础的转座子载体作为一个平台，选择性地将基因导入靶细胞。