Development of a Gene and Oligonucleotide Delivery System

基因和寡核苷酸递送系统的开发

• 批准号: 8981573
• 负责人: Gary Fujii
• 金额: $ 32.29万
• 依托单位: MOLECULAR EXPRESS, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8981573
• 关键词: Antisense Oligonucleotides; Binding; Biological Assay; Breast Cancer Cell; Capsid; Capsid Proteins; Cell Line; Cell membrane; Cells; Cellular Assay; Cytoplasm; DNA; Development; Effectiveness; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Fluorescence Microscopy; Gene Delivery; Gene Targeting; Genes; Genetic; Genetic Medicine; Healthcare; Housing; Hypoxia; Hypoxia Inducible Factor; In Vitro; Incubated; Ionic Strengths; Label; Length; Ligands; Lipid Bilayers; Lipids; Literature; MDA MB 231; Mammalian Cell; Measurement; Medical; Medicine; Methods; Modeling; Mono-S; Oligonucleotides; Organ; Oxygen; Phospholipids; Plant Viruses; Plants; Polymers; Production; Proteins; RNA; Relative (related person); Small Interfering RNA; System; Technology; Testing; Tissues; Toxic effect; Vascular Endothelial Growth Factors; Viral; Virus; Virus-like particle; Work; angiogenesis; base; cancer cell; cowpea chlorotic mottle virus; immunogenicity; interest; malignant breast neoplasm; nanoparticle; novel; novel strategies; particle; public health relevance; reconstitution; research study; self assembly; targeted delivery; tumor; uptake; vector

 DESCRIPTION (provided by applicant): The knockdown of targeted genes by anti-sense oligonucleotides (ODNS) and genetic medicines (collectively, G-MEDS) holds promise for a variety of therapies. The delivery of effective quantities of ODNS to specific cells, however, has proved to be challenging. We propose here a novel approach to ODN delivery that involves enveloped virus-like-particles (EVLPs). These delivery agents are prepared by the in vitro self- assembly of pure G-MEDS and pure viral capsid protein (CP) into virus-like nanoparticles particles (VLPs). The capsid protects the contents yet, as we have demonstrated, is capable of giving up its contents in the cytoplasm of mammalian cells. The particles, which are highly mono-disperse, are then enveloped by lipid bilayers that can suppress the immunogenicity of the VLPs and are capable of being functionalized for targeting and uptake by mammalian cells of interest. In preliminary experiments, we have demonstrated our ability to prepare EVLPs using the CP of the cowpea chlorotic mottle virus and a model antisense ODN for vascular endothelial growth factor (VEGF), a protein over-expressed in many cancer cells to stimulate angiogenesis a facilitate tumor survival under low- oxygen conditions. We propose to optimize the assembly and to functionalize the lipid bilayers with epidermal growth factor (EGF), which binds the EGF receptor that is over-expressed on cancer cells, especially those of breast cancer. The effectiveness of the EVLP will be demonstrated by assaying the reduction in the secretion of VEGF in cultured breast cancer cells.

 描述（由申请人提供）：然而，通过反义寡核苷酸（ODNS）和基因药物（统称为 G-MEDS）敲低目标基因有望将有效量的 ODNS 递送至特定细胞。 ，已被证明是具有挑战性的，我们在这里提出了一种涉及包膜病毒样颗粒（EVLP）的新 ODN 递送方法，这些递送剂是通过纯 G-MEDS 的体外自组装来制备的。和纯病毒衣壳蛋白（CP）转化为病毒样纳米粒子（VLP），衣壳可以保护其内容物，但正如我们所证明的那样，它能够放弃其在哺乳动物细胞细胞质中的内容物。单分散，然后被脂质双层包裹，可以抑制 VLP 的免疫原性，并且能够被功能化以被感兴趣的哺乳动物细胞靶向和摄取。使用豇豆褪绿斑驳病毒的CP和血管内皮生长因子（VEGF）的模型反义ODN制备EVLP，VEGF是一种在许多癌细胞中过度表达的蛋白质，可刺激血管生成并促进肿瘤在低氧条件下存活。优化组装并用表皮生长因子 (EGF) 功能化脂质双层，表皮生长因子 (EGF) 与癌细胞（尤其是乳腺癌细胞）上过度表达的 EGF 受体结合。 EVLP 将通过检测培养的乳腺癌细胞中 VEGF 分泌的减少来证明。