In Vivo Delivery of Nucleic Acids by Anionic Lipsomes

描述(由申请人提供) 目前的大量研究都是针对设计纠正或 转基因小鼠的代偿性遗传或治疗性干预 人类疾病的模型。这一领域的主要问题之一是缺乏 一种可靠的方法将遗传物质均匀地输送到血浆中 细胞膜，尤指未分裂的细胞，如神经元。我们已经开发出一种 一种可包装和输送遗传物质的阴离子脂质体 适用于有丝分裂后神经元的培养。这些阴离子脂质体进入 神经细胞的结构性内吞作用及其体外毒性的研究 属于自己的。这种新的输送系统具有实现均匀的巨大潜力 体内成熟神经元的转染法。这项拨款中提议的实验 寻求测试这种将遗传物质引入到 小鼠中枢神经系统。这些实验将为未来奠定基础 靶向a的基因和反义寡核苷酸的治疗性输送。 各种遗传性神经退行性疾病。我们的长期目标是 开发这项技术，用于实验和临床的基因操作。 我们目前的目标是测试这项技术，评估其局限性，并 优化CNS最大转染率的处方和给药方法。 实现这一目标将确定适用这一目标的最佳方案 向中枢神经系统基因传递的技术。然后，我们将能够将其应用于 三核苷酸重复病动物模型的比较 拟议的治疗亨廷顿病的基因干预和 脊髓小脑性共济失调。为了这个目标，我们建议确定DNA是否 被包裹在阴离子脂质体中可以被输送到纹状体内的细胞中 和小脑以及组织渗透和转基因的程度，可以 通过不同的中枢神经系统进入路线来实现。

DESCRIPTION (provided by applicant) A tremendous amount of current research is directed at devising corrective or compensatory genetic or therapeutic interventions in genetically altered mouse models of human disease. One of the major problems in the field is the absence of a reliable method to uniformly deliver genetic material across the plasma membrane, especially in nondividing cells such as neurons. We have developed an anionic liposomal delivery system that can package and deliver genetic material uniformly to cultures of postmitotic neurons. These anionic liposomes enter neurons via constitutive endocytosis and present no in vitro toxicity of their own. This novel delivery system holds great potential for effecting uniform transfection of mature neurons in vivo. The experiments proposed in this grant seek to test this delivery system for the introduction of genetic material into the mouse CNS. These experiments will lay the groundwork for the future therapeutic delivery of genes and antisense oligonucleotides targeted to a. variety of hereditary neurodegenerative diseases. Our long range goal is to develop this technique for both experimental and clinical genetic manipulation. Our current objective is to test this technology to assess its limits and to optimize the formulation and delivery method for maximal CNS transfection. Attainment of this objective will define the optimal protocol for applying this technology to CNS gene delivery. We will then be in a position to apply it to animal models of trinucleotide repeat diseases to compare the growing number of proposed genetic interventions for the treatment of Huntington's Disease and spinocerebellar ataxia. Towards this goal, we propose to determine if DNA encapsulated in anionic liposomes can be delivered into cells in the striatum and cerebellum and the extent of tissue penetration and transfection that can be achieved with different routes of CNS entry.