Development of Helical Polypeptides for Non-Viral Gene Delivery Systems

用于非病毒基因传递系统的螺旋多肽的开发

• 批准号: 8096083
• 负责人: Jianjun Cheng
• 金额: $ 22.46万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8096083
• 关键词: Address; Amines; Antibodies; Architecture; Binding; Blood Circulation; Cell Line; Cell Nucleus; Cells; Chemistry; Clinical Trials; Complex; Cytoplasm; Cytosol; DNA; DNA Binding; Development; Endocytic Vesicle; Environment; Equilibrium; Gene Delivery; Generations; Genetic; Genetic Materials; Goals; Health; In Vitro; Individual; Libraries; Library Materials; Lipids; Mediating; Membrane; Methodology; Nature; Non-Viral Vector; Nuclear; Nuclear Envelope; Nuclear Localization Signal; Pathway interactions; Performance; Polymers; Preparation; Property; Proteins; Reaction; Risk; Scheme; Screening procedure; Serum; Side; Small Interfering RNA; Structure; Suspension substance; Suspensions; Testing; Tissues; Vertebral column; Virus; base; design; extracellular; gene delivery system; gene therapy; immunogenic; immunogenicity; improved; in vivo; interest; meetings; monomer; non-oncogenic; non-viral gene delivery; polymerization; polypeptide; receptor; small molecule; transgene expression; uptake; vector

DESCRIPTION (provided by applicant): Because of its broad potential, gene therapy has been heavily investigated over the past 20 years. However, a clinically viable gene therapy treatment has yet to be realized. The single greatest impediment to the translational development of gene therapy is the lack of safe and efficient means to deliver genetic information to target cells and tissues. While viruses are currently the most efficient way to deliver genetic information, they also pose serious health risks-including immunogenicity and oncogenicity-which have manifested themselves in prior clinical trials. Cationic polymers and lipids have the potential to be non-immunogenic and non-oncogenic delivery vehicles, but many of the available materials are relatively inefficient. Even the most efficient lipids and polymers are orders of magnitude less efficient than viruses, often necessitating the use of micrograms of DNA to achieve transgene expression comparable to that resulting from a virus suspension containing only picograms of genetic material. In order to improve the efficiency of non-viral vectors, they must be designed to overcome extracellular barriers common to all gene delivery vehicles as well as a second set of intracellular barriers encountered once the delivery vehicle reaches the cells of interest. Specifically, 1) the vector must bind to the cell, 2) be internalized, 3) escape from endocytic vesicles into the cytoplasm, 4) move through the cytosol to the nuclear envelope, 5) migrate into the nucleus and 6) release the DNA. Viruses are very efficient because they have evolved specific functions for meeting each of these challenges. Current synthetic materials, however, lack some or all of the functions necessary for efficiently escorting the DNA from outside the extracellular environment into the nucleus. With this R21 application, we propose to develop a reactive cationic ?-helical template for the construction of a library of materials for gene delivery. We are interested in retaining helical architecture throughout the library due to the frequent occurrence of helical domains in many membrane disruptive materials. We believe having this secondary structure as the core feature of the library will yield materials which are able to effectively escape from endocytic vesicles. By subtly changing the hydrophilic/hydrophobic balance of side chains grafted onto the helical backbone, we will generate materials which have a variety of DNA binding strengths. In this manner, we hope to discover polymers with the appropriate balance of DNA binding strength and endosomolytic properties to yield efficient gene delivery. PUBLIC HEALTH RELEVANCE: This R21 application addresses endosomal escape-one of the most important barriers to efficient non-viral gene delivery-through the development of a reactive cationic ?-helical template for the construction of a library of materials for non-viral gene delivery. The cationic helical architecture is believed to aid endosomolysis while subtle changes in the hydrophilic/hydrophobic balance of side chains grafted onto the helical template will allow the identification of polymers with the appropriate balance of DNA binding strength and endosomolytic properties to yield efficient gene delivery.

描述（由申请人提供）：由于其广泛的潜力，基因治疗在过去20年中得到了大量研究。然而，临床上可行的基因疗法治疗尚未实现。基因治疗转化发展的最大障碍是缺乏安全有效的方法将遗传信息传递到靶细胞和组织。虽然病毒是目前传递遗传信息最有效的方式，但它们也会带来严重的健康风险，包括免疫原性和致癌性，这在之前的临床试验中已经表现出来。阳离子聚合物和脂质具有成为非免疫原性和非致癌性递送载体的潜力，但许多可用的材料相对低效。即使是最有效的脂质和聚合物的效率也比病毒低几个数量级，通常需要使用微克的DNA来实现转基因表达，这与仅含有皮克遗传物质的病毒悬浮液产生的转基因表达相当。为了提高非病毒载体的效率，它们必须被设计成克服所有基因递送载体共有的细胞外屏障以及一旦递送载体到达感兴趣的细胞时遇到的第二组细胞内屏障。具体地，1）载体必须结合细胞，2）被内化，3）从内吞囊泡逃逸到细胞质中，4）通过细胞质溶胶移动到核膜，5）迁移到细胞核中，以及6）释放DNA。病毒非常有效，因为它们已经进化出特定的功能来应对这些挑战。然而，目前的合成材料缺乏将DNA从细胞外环境有效护送到细胞核中所必需的一些或全部功能。通过R21的应用，我们建议开发一种反应性阳离子？用于构建基因递送材料文库的螺旋模板。由于螺旋结构域在许多膜破坏性材料中的频繁出现，我们有兴趣在整个文库中保留螺旋结构。我们相信具有这种二级结构作为文库的核心特征将产生能够有效地从内吞囊泡逃逸的材料。通过巧妙地改变接枝到螺旋骨架上的侧链的亲水/疏水平衡，我们将产生具有各种DNA结合强度的材料。通过这种方式，我们希望发现具有DNA结合强度和内体溶解性质的适当平衡的聚合物，以产生有效的基因递送。 公共卫生关系：该R21应用解决了内体逃逸-有效非病毒基因递送的最重要障碍之一-通过开发反应性阳离子？用于构建非病毒基因递送材料文库的螺旋模板。阳离子螺旋结构被认为有助于内体溶解，而接枝到螺旋模板上的侧链的亲水/疏水平衡的细微变化将允许鉴定具有DNA结合强度和内体溶解性质的适当平衡的聚合物，以产生有效的基因递送。