Histone targeted non-viral gene delivery to enhance bone repair

组蛋白靶向非病毒基因传递以增强骨修复

• 批准号: 8842129
• 负责人: Millicent O Sullivan
• 金额: $ 34.35万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8842129
• 关键词: Address; Area; BMP2 gene; Binding; Biocompatible Materials; Biology; Bone Injury; Bone Regeneration; Bone Transplantation; Cell physiology; Cells; Chemistry; Chromatin; Clinical; Clinical Research; Complex; Coupled; DNA; DNA Binding; Defect; Development; Exhibits; Fostering; Fracture; Gene Delivery; Gene Expression; Gene Transfer; Genes; Genetic Transcription; Goals; Growth Factor; Growth Factor Gene; Healed; Health; Heterotopic Ossification; Histones; Image; Image Analysis; Implant; In Situ; In Vitro; Individual; Knowledge; Letters; Link; Mediating; Mediation; Mesenchymal Stem Cells; Methods; Modeling; Monitor; Mus; Mutagenesis; Natural regeneration; Nature; Nuclear; Operative Surgical Procedures; Orthopedics; Osteogenesis; Outcome; Pathway interactions; Peptides; Post-Translational Protein Processing; Process; Production; Productivity; Proteins; Publishing; Rattus; Regulator Genes; Reporting; Role; Safety; Speed; Structure; System; Tail; Testing; Therapeutic; Tissue Engineering; Transfection; Translations; Viral; Viral Genes; base; biomaterial compatibility; bone; bone healing; bone morphogenetic protein 2; clinical efficacy; combinatorial; controlled release; cost; design; economic cost; gene delivery system; gene therapy; healing; immunogenicity; improved; in vivo; insight; interest; mimetics; nanoGold; nanoparticle; new technology; non-viral gene delivery; novel; osteogenic; plasmid DNA; repaired; scaffold; self assembly; stem cell differentiation; success; tissue repair; trafficking; uptake; viral gene delivery

DESCRIPTION (provided by applicant): Large segmental bone defects are a persistent clinical challenge that poses significant economic costs as well as costs to individual health and societal productivity. Limitations in bone grafting emphasize the need for alternative strategies. Accordingly, a myriad of therapeutic approaches have been explored to address this health problem, including the direct delivery of growth factors and delivery of viral growth factor gene therapies. While these approaches have improved healing outcomes, they have not been widely employed clinically, owing at least in part to limited protein stability (direct delivery) and safey concerns (viral gene therapies). Non-viral strategies to induce efficient, cell-mediated production of growth factors would offer a provocative approach to overcome these difficulties. We propose to address these challenges through the development of new, histone-targeted gene transfer scaffolds with tunable DNA binding and controllable gene delivery. The display of histone motifs on nanoparticle scaffolds (e.g. nanogold) will capitalize on our preliminary studies proving that post-translationally modified histone tails promote nuclear accumulation, DNA release, transcription, and enhanced transfection by non-viral vehicles. Additionally, our approach builds on established advantages of nanogold, including imaging potential, biocompatibility, functionalizability, and cell entry capacity. The novel presentation of histone tails on nanogold should mimic the native, multifaceted display and functionality of these sequences on the histone octamer. Hence, these new scaffolds should provide additional important, yet unexplored, benefits for controlling and understanding gene packaging, trafficking, and release through enhanced utilization of native gene transfer and trafficking pathways. Accordingly, the goal of this proposal is to create and optimize multifunctional "designer" histones to induce efficient gene transfer, and ultimately, enable improved tissue repair for orthopedics and other applications. We will produce nanogold-plasmid DNA (pDNA) assemblies, and will determine whether the multivalent presentation of pDNA-binding residues and histone peptides enhances pDNA-binding stability and improves transfection. We will capitalize on traditional as well as nanogold-specific imaging analyses to elucidate key steps in non-viral gene transfer, and will clearly link enhanced gene transfer efficiency to improved osteogenic potential of growth factors such as bone morphogenetic protein-2 (BMP-2). Finally, we will use murine and rat orthopedic models to test the ability of the BMP-2 gene product to enhance bone formation and increase the speed and efficacy of defect repair. Our approaches will not only elucidate mechanistic details about the trafficking of histone-associated genes, but will also ultimately will be useful as a general biomaterials platform applicable to bone repair, implant functionalization, and tissue engineering.

描述（由申请人提供）：大节骨缺陷是持续的临床挑战，其构成了巨大的经济成本以及个人健康和社会生产力的成本。骨嫁接的局限性强调了替代策略的需求。因此，已经探索了无数的治疗方法来解决这个健康问题，包括直接递送生长因子和病毒生长因子基因疗法的递送。尽管这些方法改善了愈合结果，但尚未在临床上广泛使用，至少部分是由于有限的蛋白质稳定性（直接递送）和SAFEY关注（病毒基因疗法）。非病毒策略诱导有效的，细胞介导的产生 增长因素将提供一种挑衅的方法来克服这些困难。 我们建议通过开发具有可调DNA结合和可控基因递送的新的，组蛋白的基因转移支架来应对这些挑战。在纳米颗粒支架上的组蛋白基序的显示（例如纳米卵）将利用我们的初步研究 证明，翻译后修饰的组蛋白尾巴促进了核积累，DNA释放，转录和通过非病毒车辆的转染增强。此外，我们的方法基于纳米质量的既定优势，包括成像潜力，生物相容性，功能化性和细胞进入能力。组蛋白尾巴在纳米质量上的新型表现应模仿组蛋白八聚体上这些序列的天然，多方面的显示和功能。因此，这些新的脚手架应通过通过增强本地基因转移和运输途径的利用来控制和理解基因包装，运输和释放的其他重要但未开发的好处。 因此，该提案的目的是创建和优化多功能的“设计师”组蛋白来诱导有效的基因转移，并最终为骨科和其他应用提供改进的组织修复。我们将产生纳米质质粒DNA（pDNA）组件，并确定pDNA结合残基和组蛋白肽的多价表示是否增强了pDNA结合稳定性并改善转染。我们将利用传统和纳米质量特异性的成像分析，以阐明非病毒基因转移的关键步骤，并将明确将增强的基因转移效率与改善生长因子（例如骨形态发生蛋白2（BMP-2））的成骨潜力联系起来。最后，我们将使用鼠和大鼠骨科模型来测试BMP-2基因产物增强骨形成并提高缺陷修复的速度和功效的能力。我们的方法不仅将阐明有关组蛋白相关基因的运输的机理细节，而且最终也将作为适用于骨修复，植入物功能化和组织工程的一般生物材料平台有用。