Controlled Release of DNA from Surfaces

DNA 从表面的受控释放

• 批准号: 7463115
• 负责人: DAVID M LYNN
• 金额: $ 32.06万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7463115
• 关键词: Address; Adverse effects; Animal Model; Antineoplastic Agents; Architecture; Arteries; Behavior; Biological Assay; Biomedical Research; Blood Vessels; Cells; Charge; Clinic; Clinical; Complex; Coronary Arteriosclerosis; DNA; DNA Structure; DNA delivery; Deposition; Development; Drug Delivery Systems; Engineering; Environment; Esters; Evaluation; Family suidae; Film; Fluorescence; Foundations; Future; Gene Delivery; Genes; Gray unit of radiation dose; Health; Human; Hydrophobicity; Implant; In Vitro; Investigation; Ions; Lead; Life; Localized; Mammalian Cell; Mechanics; Mediating; Methods; Morphology; Organic solvent product; Oryctolagus cuniculus; Outcome; Patients; Pharmaceutical Preparations; Physiological; Placement; Polyamines; Polymers; Procedures; Process; Property; Public Health; Range; Rate; Reporter; Reporter Genes; Research; Scheme; Serum; Side; Solid; Solutions; Spatial Distribution; Stainless Steel; Stents; Structure; Surface; Sus scrofa; Testing; Therapeutic; Thick; Tissues; Transfection; Vertebral column; analytical method; base; biodegradable polymer; cell type; controlled release; density; design; enhanced green fluorescent protein; iliac artery; implantable device; improved; in vivo; innovation; plasmid DNA; polyanion; polycation; pre-clinical; research clinical testing; research study; restenosis; therapeutic gene; tool; trafficking

DESCRIPTION (provided by applicant): This proposal seeks to develop new materials and approaches to the design of ultrathin films that permit spatial and temporal control over the release of DNA from surfaces. The proposed research seeks to test the hypothesis that ultrathin multilayered films composed of alternating layers of plasmid DNA and degradable polyamines can be used to promote broad and tunable control over surface-mediated cell transfect ion in vitro and in vivo. Our approach is founded on preliminary observations that ultrathin films (e.g., 100 nm thick) fabricated from alternating layers of DNA and degradable cationic polymers promote cell transfect ion when placed in contact with cells and tissues. These critical observations suggest the basis of general methods that permit spatial and temporal control over the release of DNA in vitro and facile new methods for the coating and localized delivery of DNA from the surfaces of indwelling devices in vivo. Our preliminary investigations validate the principles required to exploit ultrathin multilayered films for the surface-mediated delivery of DNA. The following Specific Aims are designed to systematically evaluate our hypothesis and evaluate the full potential of this approach. They are: 1) To characterize the influence of polymer structure on rates of release of DNA and the structure and morphology of DNA released from ultrathin films fabricated from alternating layers of plasmid DNA and degradable polyamines; 2) To investigate the influence of polymer structure and film architecture on levels of cell transfect ion and viability in vitro when objects coated with ultrathin films are placed in contact with cells; and 3) To demonstrate that materials developed and evaluated under Aims 1 and 2 can be used to coat indwelling devices such as intravascular stents, and that film-coated stents can be used transfect vascular tissue in vivo when implanted in the arteries of rabbits and pigs. We envisage the outcomes of the proposed research as having substantial fundamental and applied impacts by (i) providing new materials and tools for basic biomedical research and surface-mediated transfection in vitro, (ii) by developing robust new methods for the localized delivery of DNA from indwelling devices in vivo, and (iii) by providing a foundation for future research and the selection of materials with properties (e.g., specific release profiles) suited for the evaluation of therapeutic gene candidates directed toward a broad range of different therapeutic outcomes. Public Health Relevance: The potential impact of methods that provide control over the release of DNA from surfaces ranges from applications in basic biomedical research to, ultimately, the realization of gene-based therapies in the clinic. The research described in this proposal will impact public health by providing new tools for surface-mediated DNA delivery in vitro and new methods for localized delivery of DNA from indwelling devices in vivo that could improve the efficacies and reduce the side effects of gene-based therapies.

描述（由申请人提供）：该提案旨在开发新材料和超薄膜设计方法，以允许对DNA从表面的释放进行空间和时间控制。拟议的研究旨在测试的假设，即交替层的质粒DNA和可降解的多胺组成的多层膜可用于促进广泛的和可调的控制在体外和体内的表面介导的细胞粘附离子。我们的方法是建立在初步观察的基础上的，100 nm厚），其由DNA和可降解阳离子聚合物的交替层制成，当与细胞和组织接触时促进细胞粘附。这些重要的观察结果表明，一般的方法，允许在体外释放的DNA的空间和时间控制和简便的新方法的涂层和局部交付的DNA从留置装置的表面在体内的基础。我们的初步研究验证了利用多层膜表面介导的DNA传递所需的原则。以下具体目标旨在系统地评估我们的假设，并评估这种方法的全部潜力。它们是：1）表征聚合物结构对DNA释放速率的影响以及从由质粒DNA和可降解多胺交替层制成的聚合物膜释放的DNA的结构和形态：2）研究聚合物结构和膜结构对当将涂覆有聚合物膜的物体与细胞接触时的体外细胞粘附水平和存活率的影响;和3）证明根据目标1和2开发和评价的材料可用于涂覆留置器械（如血管内支架），并且当植入兔和猪的动脉时，薄膜涂层支架可用于体内切除血管组织。我们设想所提出的研究的结果具有实质性的基础和应用影响，通过（i）为基础生物医学研究和体外表面介导的转染提供新的材料和工具，（ii）通过开发用于体内留置装置的DNA局部递送的稳健的新方法，以及（iii）通过为未来的研究和具有特性的材料的选择提供基础（例如，特异性释放曲线），适用于评价针对广泛的不同治疗结果的治疗性基因候选物。 公共卫生相关性：控制DNA从表面释放的方法的潜在影响范围从基础生物医学研究中的应用到最终在临床中实现基于基因的治疗。本提案中描述的研究将通过提供体外表面介导DNA递送的新工具和体内留置装置局部递送DNA的新方法来影响公共卫生，这些方法可以提高疗效并减少基于基因的治疗的副作用。