Reversing Electrostatic Interactions for Improved Gene Delivery

逆转静电相互作用以改善基因传递

• 批准号: 7261234
• 负责人: MARK W. GRINSTAFF
• 金额: $ 34.86万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-14 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7261234
• 关键词: Address; Affect; Animals; Benchmarking; Binding; Biological; Biological Assay; Breast Cancer Cell; Breast Cancer Treatment; Cancer cell line; Caring; Cells; Characteristics; Charge; Chemicals; Chemistry; Choline; Cleaved cell; Clinical; Combined Modality Therapy; Covalent Interaction; DNA; DNA Binding; Data; Dendrimers; Differential Scanning Calorimetry; Disease; Electrostatics; Endocytosis; Engineering; Esters; Esthetics; Fluorescence; Fluorescence Resonance Energy Transfer; Formazans; Future; Galactosidase; Gene Delivery; Gene Proteins; Genes; Goals; Hydrogen Bonding; In Vitro; Investigation; Label; Lead; Lipid Bilayers; Lipids; Mechanics; Medicine; Membrane; Microscopic; Microscopy; Molecular; Molecular Structure; Nature; Non-Viral Vector; Nuclear; Nucleic Acids; Numbers; Optics; Pathway interactions; Phosphorylcholine; Play; Polymers; Process; Property; Protein p53; Proteins; Range; Rate; Reaction; Reporter Genes; Research; Research Personnel; Role; Standards of Weights and Measures; Structure; Surface; System; TP53 gene; Techniques; Today; Transfection; Translating; Transmission Electron Microscopy; Tumor Suppressor Genes; Vesicle; X ray diffraction analysis; X-Ray Diffraction; base; cell type; cytotoxicity; design; driving force; esterase; gene therapy; improved; in vivo; malignant breast neoplasm; millimeter; nanoscale; novel strategies; programs; research clinical testing; response; small molecule; synthetic construct; therapeutic gene; vector

DESCRIPTION (provided by applicant): The overall goal of the proposed effort is to use the principles of intermolecular forces to design improved cationic amphiphile/DNA supramolecular assemblies for gene delivery. Specifically, we will design, synthesize, and evaluate new charge-reversal amphiphiles for gene delivery. These functional amphiphiles undergo an electrostatic transition from cationic to anionic in cells to release DNA from the supramolecular assembly. We hypothesize that this change in electrostatic interactions with DNA will translate to enhanced gene transfection efficiency. A detailed mechanistic investigation is proposed which entails the following three specific aims for this four-year proposal: Aim 1. Determine the key molecular characteristics of the amphiphile required for 1) DNA binding and release of DNA from the supramolecular assembly and 2) destabilization of lipid bilayers. Aim 2. Characterize the charge-reversal amphiphiles and amphiphile/DNA supramolecular assemblies. Aim 3. Evaluate functional interactions of charge-reversal amphiphile/DNA supramolecular assemblies with cells in vitro and deliver the p53 gene to breast cancer cells. Completion of these aims will afford: (1) the identification of one or more charge-reversal amphiphiles that can deliver DNA to cells; (2) the mechanism of nucleic acid delivery with these charge-reversal amphiphiles; (3) an understanding of the advantages and limitations of these amphiphiles for gene delivery in a given cell type; (4) the demonstration of a new approach that is a conceptual departure from the current gene delivery vectors; (5) delivery of the p53 gene to breast cancer cells; and (6) potentially identify the nature of the rate- limiting step in gene delivery. Breast cancer is one disease that would benefit from improved or alternative treatment options. Today, there is no standard of care for metastatic breast cancer; all of the first-line combination therapies are regarded as equally efficacious at about a 60% response rate. Thus, we are focusing on the delivery of the tumor suppressor gene (p53) for the treatment of breast cancer.

描述（由申请人提供）：拟议工作的总体目标是使用分子间力原理来设计用于基因传递的改进的阳离子两亲体/DNA超分子组装体。具体来说，我们将设计、合成和评估用于基因传递的新型电荷反转两亲体。这些功能性两亲体在细胞中经历从阳离子到阴离子的静电转变，从超分子组装中释放DNA。我们假设这种与DNA静电相互作用的变化将转化为提高基因转染效率。建议进行详细的机械调查，为这项为期四年的建议确定以下三个具体目标：目标1。确定两亲体所需的关键分子特征：1)DNA结合和从超分子组装中释放DNA； 2)脂质双分子层的不稳定。目标2。表征电荷反转两亲体和两亲体/DNA超分子组合。目标3。在体外评估电荷反转两亲体/DNA超分子组件与细胞的功能相互作用，并将p53基因传递给乳腺癌细胞。完成这些目标将提供：(1)鉴定一种或多种可以向细胞传递DNA的电荷反转两亲动物；(2)这些电荷反转的两亲体传递核酸的机制；(3)了解这些两亲体在特定细胞类型中传递基因的优势和局限性；(4)展示一种新方法，这种方法在概念上与目前的基因传递载体有所不同；(5)向乳腺癌细胞传递p53基因；(6)潜在地确定基因传递中速率限制步骤的性质。乳腺癌是一种将受益于改进或替代治疗方案的疾病。今天，对于转移性乳腺癌没有标准的治疗方法；所有的一线联合治疗被认为同样有效，有效率约为60%。因此，我们将重点放在肿瘤抑制基因（p53）的传递上，以治疗乳腺癌。