MUCOSAL GENE TRANSFER USING DENDRIMER POLYMERS

最近的工作证明了聚酰胺胺树状大分子，一种新型的 高度支化的树枝状聚合物，可以用来调节有效的 将遗传物质转移到真核细胞。这种聚合物有一种 高度明确的结构，是惰性的和非免疫原性的，并结合细胞核 在各种条件下的任何大小或结构的酸。高 高效的DNA转移已经被介导到各种各样的细胞中 使用树枝状大分子/DNA复合体进行体外实验。这些细胞包括呼吸道 上皮细胞系。此外，初步证据表明， DNA/树枝状大分子复合体可在体内转染肺细胞，DNA 与树枝状大分子形成络合物后，在体内具有较长的存留时间。此外, 改变树枝状大分子结构中特定参数的独特能力 允许将这些分子用作探针来检查特定方面 不同类型的呼吸道上皮细胞的转染率。这个 这一提议的中心假设是树突的独特形式 聚合物将结合并有效地传递治疗性核酸 分子在体内对呼吸道粘膜细胞的作用。这种形式的基因转移 将被用于治疗呼吸道炎症疾病和 涉及将不同形式的治疗性核酸输送到 改变各种炎症事件。首先，一些不同的 将生产树枝状大分子以确定聚合物设计中的哪些变化 或聚合物与分子的结合，如碳水化合物、药物、 抗体，或合成肽，将允许有效的靶向和 DNA/树枝状大分子复合体转移到特定的呼吸道上皮细胞或 炎性细胞。一个明确的肺纤维化模型将在那时 被用来测试基因干预的有效性 DNA/树枝状大分子复合体可预防纤维化的发展。这 提案是基于我的实验室的工作和来自其他人的研究 已经证明树枝状大分子很容易连接到 靶向分子和共轭树枝状大分子可以有效靶向 放射性核苷酸对体内细胞的影响。这些研究的结果将 提供有关基因传递的基本信息，以便进一步 支气管腔内基因转移方案和试剂的改进。

Recent work in has demonstrated the plyamidoamine dendrimers, a new type of highly branched dendtiric polymer, can b used to mediate the efficient transfer of genetic material into eukaryotic cell. This polymer has a highly defined structure, is inert and non-immunogenic, and binds nucleic acids of any size or structure in a wide variety of conditions. High efficiency DNA transfer has been mediated to a wide variety of cells in vitro using the dendrimer/DNA complexes. These cells have included airway epithelial cells lines. In addition, preliminary evidence suggests that DNA/dendrimer complexes can transfect lung cells in vivo and that DNA complexed to dendrimers has prolonged persistence in vivo. In addition, the unique ability to alter specific parameters in dendrimer structure allows these molecules to be used as probes to examine particular aspects of the transfection of different types of airway epithelial cells. The central hypothesis of this proposal is that unique forms of dendritic polymers will bind and effectively deliver therapeutic nucleic acid molecules to airway mucosal cells in vivo. This form of gene transfer would be employed for the treatment of airway inflammatory diseases an involve the delivery of different forms of therapeutic nucleic acids to alter a variety of inflammatory events. First, a number of different dendrimers will be produced to determine what alterations in polymer design or the conjugation of the polymers to molecules, such carbohydrates, drugs, antibodies, or synthetic peptides, will allow the efficient targeting and transfer of DNA/dendrimer complexes to specific airway epithelial or inflammatory cells. A well defined model of pulmonary fibrosis will then be employed to test the efficacy of genetic intervention using DNA/dendrimer complexes to prevent the development of fibrosis. This proposal is based on work from my laboratory and also studies from other groups that have documented that dendrimers are easily conjugated to targeting molecules and that conjugated dendrimers can efficiently target radionucleotides to cells in vivo. The results of these studies will provide basic information about gene delivery that should allow further refinements in protocols and agents for endobronchial gene transfer.