Engineering functionalized diamond nanoparticles for intracellular delivery of nucleic acids

工程化功能化金刚石纳米粒子用于核酸的细胞内递送

• 批准号: RGPIN-2015-03689
• 负责人: Badea, Ildiko
• 金额: $ 2.19万
• 依托单位: University of Saskatchewan
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=683448
• 关键词: Engineering; functionalized; diamond; nanoparticles; intracellular

The objective of my research program is to engineer novel nanomaterials that carry nucleic acids into mammalian cells. Gene delivery has been a significant research area of interest for the past two decades, but the mainstream chemical gene delivery nanomaterials, lipids and polymers pose limitations with respect to their stability, efficiency and toxicity. My research program focuses on the use of solid-core nanoparticles, such as nanodiamonds, to overcome these barriers. Nanodiamonds have advantages over lipid and polymer nanoparticles by virtue of a uniform size distribution and a large surface area available for chemical modifications. They are nontoxic in doses necessary for gene delivery.****My research strategy is to chemically link amino acids to the nanodiamond in order to facilitate their interaction with nucleic acids. The novelty of this program lies in the approaches to the synthesis of nanoparticles conjugated with bioactive molecules, amino acids.****The short-term research objectives build on my previous work: design of self-assembling lipid-based and diamond-based nanoparticles. To achieve our goal, we will chemically link amino acids on the surface of the nanodiamond resulting in functionalized nanoparticles with surface properties amenable to binding nucleic acids. We will assess the ability of these functionalized nanoparticles to deliver the bound nucleic acids into model mammalian cells. The pathways of cellular uptake and the mechanism of potential toxicity will be investigated. Relationships between structure/architecture - delivery efficiency/toxicity will be established. This will lead to the identification of candidate vectors with optimal properties (improved binding properties, stability, and cellular uptake with no or minimal toxicity) for delivering nucleic acids into cells.****To study the architecture of the delivery systems, high-resolution microscopy, flow cytometry and synchrotron-based techniques will be used. Correlating the surface properties of these nanoparticles with the delivery efficiency will allow for mapping universal and carrier-specific delivery mechanisms.****On the long term, we will explore whether the basic knowledge on chemical functionalization of the nanodiamond particles could be expanded to other solid-core and soft nanoparticles, establishing universal delivery principles for nucleic acids. This research program will advance knowledge of the use and safety of the functionalized diamond-core nanoparticles as delivery systems for nucleic acids. Optimization of the characteristics of these delivery systems will benefit nanotechnology drug delivery (a global market of $1.2 billion with an estimated growth rate of 49% in the next decade), eventually leading to gene therapy in humans and animals. Understanding the interactions of nanodiamonds with nucleic acids can also translate into biosensor applications.******

我的研究计划的目标是设计新型纳米材料，将核酸带入哺乳动物细胞。在过去的二十年里，基因传递一直是一个重要的研究领域，但主流的化学基因传递纳米材料、脂类和聚合物在稳定性、有效性和毒性方面存在局限性。我的研究计划重点是使用固体核心纳米颗粒，如纳米钻石，以克服这些障碍。纳米钻石比脂类和聚合物纳米粒子具有更大的优势，因为纳米钻石具有均匀的尺寸分布和较大的可用于化学修饰的表面积。它们在基因传递所需的剂量上是无毒的。*我的研究策略是将氨基酸化学连接到纳米钻石上，以促进它们与核酸的相互作用。这项计划的新颖性在于合成与生物活性分子、氨基酸结合的纳米粒子的方法。*短期研究目标建立在我之前的工作基础上：设计基于脂类和钻石的自组装纳米粒子。为了实现我们的目标，我们将把纳米钻石表面的氨基酸化学连接起来，形成具有表面属性的功能化纳米粒子，这些纳米粒子的表面性质符合结合核酸的要求。我们将评估这些功能化纳米颗粒将结合的核酸输送到模型哺乳动物细胞的能力。将研究细胞摄取的途径和潜在毒性的机制。将建立结构/架构-传递效率/毒性之间的关系。这将导致进一步确定具有最佳性质(改进的结合特性、稳定性和细胞摄取，没有或最小毒性)的候选载体，以将核酸输送到细胞中。*为了研究输送系统的结构，将使用高分辨率显微镜、流式细胞仪和基于同步加速器的技术。将这些纳米颗粒的表面性质与输送效率相关联，将允许绘制通用的和载体特定的输送机制。*从长远来看，我们将探索纳米金刚石颗粒的化学功能化的基本知识是否可以扩展到其他固体核和软纳米颗粒，建立核酸的通用输送原则。这项研究计划将促进人们对功能化钻石核纳米颗粒作为核酸输送系统的使用和安全性的了解。这些输送系统的特性的优化将使纳米技术药物输送受益(全球市场规模为12亿美元，未来十年的增长率估计为49%)，最终导致人类和动物的基因治疗。了解纳米钻石与核酸的相互作用也可以转化为生物传感器的应用。