FUNCTIONAL AND TARGETING POLYMERIC GENE CARRIERS

描述(改编自申请人的摘要)：在本申请中， 研究人员提议设计两种新的聚合基因载体。第一 载体是一种树枝状聚合物，由聚乙二醇(PEG)和 聚(L-赖氨酸)PLL(聚乙二醇b-PLL)，水溶性，无毒。它形成了 与质粒DNA的自组装复合体。第二个载体是聚乙二醇接枝的 PLL与动脉壁靶向部分(靶向 分子-聚乙二醇-g-PLL)。靶向部分由多肽或蛋白质组成 低密度脂蛋白(LDL)中的(ApoB-100)配体。提高工作效率 ，研究人员还将整合一种 内体干扰肽(EDP)与核定位信号(NLS) 多肽分别与树枝状大分子反应。在系统中优化后 研究，这两种新型基因载体系统将在两个方面进行进一步的评价 特征明确的心血管疾病模型(缺血性心脏病(IHD) 和血管成形术导致的再狭窄)，目前最有效的治疗方法有四种 基因(用于IHD的血管内皮生长因子、成纤维细胞生长因子-2和eNOS，截短的转化生长因子-β受体II (TTbetaR-II用于再狭窄)。尽管编码血管内皮生长因子和蛋白质的裸质粒 成纤维细胞生长因子-2已被证明通过直接心肌发挥治疗作用。 注射，裸露的DNA绝不是一种有效的基因传递方式。这个 过去心血管疾病基因治疗的失败是由于缺乏 合适的基因转移载体。调查人员认为，拟议的 基因载体在以下方面优于裸DNA和脂质体等其他载体 将感兴趣的基因传递到动脉壁细胞和心肌。为 再狭窄基因治疗的研究，使用局部基因输送装置 提出了一种应用于血管壁的动物模型。为了预防 再狭窄形成，将选择猪冠状动脉损伤模型 为了评估我们新基因载体的有效性，治疗性基因， 基因输送导管对预防再狭窄形成的作用。这 项目的设计是为了让我们的新基因载体和他们适当的 给药方式可调整为临床使用。这些研究将提供 重要信息，不仅是为了进一步成功的治疗和预防 对人类IHD和再狭窄的研究，也用于在其他疾病中使用基因治疗 疾病状态。

DESCRIPTION (Adapted from the Applicant's Abstract): In this application, the investigators propose to design two new polymeric gene carriers. The first carrier is a dendrimer polymer consisting of polyethylene glycol (PEG) and poly(L-lysine) PLL (PEG-b-PLL) that is water-soluble and non-toxic. It forms self-assembling complexes with plasmid DNA. The second carrier is PEG grafted PLL conjugated with an artery wall-targeting moiety (targeting molecule-PEG-g-PLL). The targeting moiety consists of a peptide or protein (apoB-100) ligand in low-density lipoprotein (LDL). To enhance the efficiency of plasmid DNA transfection, the investigators will also incorporate an endosomal disrupting peptide (EDP) and a nuclear localization signal (NLS) peptide to the dendrimers, respectively. After optimization in systemic studies, these two novel gene carrier systems will be further evaluated in two well characterized cardiovascular disease models (ischemic heart disease (IHD) and angioplasty induced restenosis) with four currently most potent therapeutic genes (VEGF, FGF-2 for IHD, and eNOS, truncated TGF-beta receptor II (TTbetaR-II) for restenosis). Although naked DNA plasmids encoding VEGF and FGF-2 have shown to be therapeutically effective by direct myocardium injection, naked DNA is by no means an efficient mode of gene delivery. The past failures in cardiovascular disease gene therapy are due to a lack of appropriate gene transfer carriers. The investigators believe that the proposed gene carriers are superior to naked DNA and other carriers such as liposomes in delivering the gene of interest to artery wall cells and the myocardium. For the study of restenosis gene therapy, the use of a local gene delivery device applied to vascular wall of an animal model is proposed. For prevention of restenosis formation, a porcine coronary artery injury model will be selected to evaluate the effectiveness of our new gene carriers, the therapeutic genes, and the gene delivery catheters on prevention of restenosis formation. This project is designed so that our new gene carriers and their proper administration can be adapted for clinical use. These studies will provide important information, not only for further successful treatment and prevention of human IHD and restenosis, but also for the use of gene therapy in other disease states.