Collaborative Research: An Experimental/Theoretical Program on Reconfigured Polycationic Architectures for Improved Gene Therapy

合作研究：用于改进基因治疗的重构聚阳离子结构的实验/理论计划

• 批准号: 1460380
• 负责人: Arthi Jayaraman
• 金额: $ 7.94万
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1460380&HistoricalAwards=false
• 关键词: Collaborative; Research; Experimental; Theoretical; Program

This collaborative research award by the Biomaterials program in the Division of Materials Research, made to the University of Massachusetts and University of Colorado, is to carry out experiments and simulations to understand polycation-DNA complexation towards improved non-viral gene therapy using synthetic polymers. This award cofounded by the Polymer program in the Division of Materials Research seeks to understand DNA complexation with tailored macromolecular cationic architectures, and the impact of different modes of complexation on DNA delivery. The proposed experiments focus on synthetic polycations, in which cationic oligomers are grafted pendent to a polymer backbone at precise inter-graft spacing. This architecture platform will be augmented by insertion of nuclear localization sequences into the backbone in preferred orientations. Polyplex stability and DNA delivery efficiency will be determined in cell culture under a variety of conditions, aiming to combine high cell viability with excellent transfection efficiency. The computational approach, carried out in conjunction with experiments, will involve: 1) atomistic simulations to explain the effects of systematically varying architectural and chemical features of the polycation on free energy of DNA-polycation binding, and in turn connect to experimental trends in polyplex stability; and 2) coarse-grained simulations that focus on polyplex structure (relative size, shape and charge) as a function of polycation architecture and composition, which directly relate to experiments on polyplex structure, and carries key implications for serum stability and transfection efficiency. In addition to transforming the understanding and practice of polymer-based gene therapy, the project will open a unique opportunity to train graduate students in an integrated multidisciplinary experimental/theoretical project through regular inter-group interactions via SkypeTM sessions, and by devoting time and resources for the researchers to visit their collaborator's institution. The PIs will maintain a commitment to participation of undergraduates in this research project, utilizing the Undergraduate Research Opportunity and Research Experience for Undergraduates programs at University of Colorado and University of Massachusetts. Planned workshop activities would connect U.S. and worldwide leaders in both experiment and computation of polyelectrolytes and complexation, and would involve young scientists at the outset of their careers, through the organization of meetings at the Telluride Science Research Conference, a highly interactive setting for research presentations and discussions. The design and implementation of effective polymer based gene delivery will advance genomic research and open new avenues to deliver DNA for treating many diseases, such as muscular dystrophy. There is a pressing need for improved DNA delivery, since currently used delivery methods, based on viruses, have their own health and safety risks. In this collaborative project, computer simulations will help experimentalists decide which polymers to prepare for complexing DNA, and how to modify those polymers for most effective therapeutic action. Outcomes of this project include both the development of novel and more efficient DNA delivery agents, and a more thorough fundamental understanding of charged polymers and their interactions with key therapeutic biomolecules such as DNA and RNA. The multidisciplinary nature of this experimental/theoretical project will allow for a well-rounded training of students in Massachusetts and Colorado, at the high school, undergraduate, and graduate levels, in areas of molecular simulation, polymer synthesis, and gene delivery.

这项合作研究奖由材料研究部生物材料项目授予马萨诸塞大学和科罗拉多大学，旨在进行实验和模拟，以了解聚阳离子-DNA 络合，从而使用合成聚合物改进非病毒基因治疗。该奖项由材料研究部聚合物项目共同创立，旨在了解 DNA 与定制大分子阳离子结构的络合，以及不同络合模式对 DNA 传递的影响。所提出的实验重点是合成聚阳离子，其中阳离子低聚物以精确的接枝间间距接枝到聚合物主链上。该架构平台将通过将核定位序列以首选方向插入主干来增强。 Polyplex 的稳定性和 DNA 递送效率将在多种条件下的细胞培养中测定，旨在将高细胞活力与出色的转染效率结合起来。与实验结合进行的计算方法将涉及：1）原子模拟，以解释系统地改变聚阳离子的结构和化学特征对DNA-聚阳离子结合自由能的影响，并进而与聚合复合物稳定性的实验趋势联系起来； 2) 粗粒度模拟，重点关注多聚复合物结构（相对大小、形状和电荷）作为聚阳离子结构和组成的函数，这与多聚复合物结构实验直接相关，并对血清稳定性和转染效率具有重要影响。除了改变对基于聚合物的基因治疗的理解和实践之外，该项目还将提供一个独特的机会，通过 SkypeTM 会议定期进行小组间互动，并为研究人员投入时间和资源来参观其合作者的机构，从而在综合多学科实验/理论项目中培训研究生。 PI 将利用科罗拉多大学和马萨诸塞大学的本科生研究机会和本科生研究经验项目，继续承诺让本科生参与该研究项目。计划中的研讨会活动将通过在特柳赖德科学研究会议上组织会议，将聚电解质和络合实验和计算方面的美国和世界领先者联系起来，并让年轻科学家参与到职业生涯的开始，特柳赖德科学研究会议是一个高度互动的研究演示和讨论环境。 基于聚合物的有效基因传递的设计和实施将推进基因组研究，并开辟新的途径来传递 DNA 以治疗许多疾病，例如肌营养不良症。由于目前使用的基于病毒的递送方法有其自身的健康和安全风险，因此迫切需要改进 DNA 递送。在这个合作项目中，计算机模拟将帮助实验人员决定准备哪些聚合物来复合 DNA，以及如何修改这些聚合物以获得最有效的治疗作用。 该项目的成果包括开发新型且更高效的 DNA 递送剂，以及对带电聚合物及其与 DNA 和 RNA 等关键治疗生物分子的相互作用有更全面的基础了解。该实验/理论项目的多学科性质将使马萨诸塞州和科罗拉多州的高中、本科生和研究生水平的学生在分子模拟、聚合物合成和基因传递领域得到全面的培训。