Efficient and Selective Delivery of Oligonucleotides

寡核苷酸的高效和选择性递送

• 批准号: 6938563
• 负责人: CHARLES M. ROTH
• 金额: $ 20.07万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6938563
• 关键词: DNA; antisense nucleic acid; biological transport; biomimetics; cell membrane; cell type; cytotoxicity; fibroblasts; functional /structural genomics; galactose; gene delivery system; gene expression; gene therapy; glycopeptides; ligands; liver cells; mathematical model; messenger RNA; nucleic acid metabolism; oligonucleotides; polymerase chain reaction; polymers; transfection /expression vector

The ability to modulate cell behavior through genetic modification has great potential as a therapeutic strategy, as well as providing a powerful tool for elucidating gene function (so-called functional genomics). Antisense oligonucleotides, which are most commonly single-stranded DNA molecules 15-25 nucleotides in length, modulate gene expression by binding to a complementary segment on the mRNA from the target gene. While antisense technology is becoming a viable therapeutic entity and platform for functional genomics, a major barrier to its widespread practice still exists: the delivery of the genetic material (polynucleic acid) to cells in a quantity that is biologically effective and in a form that is functionally intact, yet non-toxic. Our overall goal is to deliver antisense molecules selectively to a target cell type (hepatocytes), resulting in low-dose inhibition of expression of genes of interest. To achieve this goal, we will develop a new family of multifunctional DNA delivery vectors (multiplexes) using a combinatorial synthesis approach. These vectors will possess biomimetic polymers that condense DNA, cationic peptides that destabilize cellular membranes, and galactose moieties that target them to hepatocytes (primary liver cells). Vectors will be characterized for size, stability, and cytotoxicity. We will study the adsorption of these materials to target vs. non-target cells, and interpret the results in the framework of a colloid-chemical mathematical model, which will be used to refine and optimize the composition of the vectors. The effectiveness of these vectors to deliver gene expression-modulating antisense oligonucleotides will be evaluated and assessed to further refine the approach. W expect the long-term outcome to be a selective and efficient method for oligonucleotide delivery for therapeutic and functional genomics applications.

通过基因修饰调节细胞行为的能力作为一种治疗策略具有巨大的潜力，并为阐明基因功能（所谓的功能基因组学）提供了强大的工具。 反义寡核苷酸最常见的是长度为 15-25 个核苷酸的单链 DNA 分子，通过与靶基因 mRNA 上的互补片段结合来调节基因表达。 虽然反义技术正在成为功能基因组学的可行治疗实体和平台，但其广泛应用的主要障碍仍然存在：将遗传物质（多核酸）以生物有效的量、功能完整且无毒的形式递送至细胞。我们的总体目标是将反义分子选择性地递送至靶细胞类型（肝细胞），从而低剂量抑制感兴趣基因的表达。 为了实现这一目标，我们将使用组合合成方法开发一个新的多功能 DNA 递送载体（多重）家族。 这些载体将拥有浓缩 DNA 的仿生聚合物、破坏细胞膜稳定性的阳离子肽以及将其靶向肝细胞（原代肝细胞）的半乳糖部分。 将表征载体的大小、稳定性和细胞毒性。 我们将研究这些材料对靶细胞和非靶细胞的吸附，并在胶体化学数学模型的框架中解释结果，该模型将用于细化和优化载体的组成。 将评估这些载体递送调节基因表达的反义寡核苷酸的有效性，以进一步完善该方法。 我们期望长期结果是一种用于治疗和功能基因组学应用的选择性且有效的寡核苷酸递送方法。