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-2020-05315
• 负责人: Badea, Ildiko
• 金额: $ 2.04万
• 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: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=740504
• 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 for the past two decades, but the mainstream 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 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 and characterization of nanoparticles conjugated with bioactive molecules, amino acids and elucidation of their beneficial and detrimental interactions with mammalian cells, organs and whole body. 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 decorate the surface of nanodiamond with amino acids resulting in functionalized nanoparticles amenable to binding nucleic acids. We will assess the ability of these functionalized nanoparticles to deliver nucleic acids into model cells. The biodistribution of various functionalized nanodiamonds will be monitored. Relationships between structure/architecture - delivery efficiency/toxicity will be established. This will lead to the selection of candidate vectors with optimal properties (improved binding properties, cellular uptake and high activity with no or minimal toxicity, and stability in biological environment) 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. Long term, we will explore how 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. Correlation of biodistribution to surface properties will be used as a model for predicting the biological fate of other man-made nanoparticles with similar surface properties. 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 (a global market of $1.2B), eventually leading to gene therapy in humans and animals.

我的研究项目的目标是设计新型纳米材料，将核酸带入哺乳动物细胞。基因递送在过去的二十年中一直是一个重要的研究领域，但主流的基因递送纳米材料、脂质和聚合物在其稳定性、效率和毒性方面存在局限性。我的研究项目集中在使用固体核心纳米粒子，如纳米金刚石，以克服这些障碍。纳米金刚石具有均匀的尺寸分布和可用于化学改性的大表面积。它们在基因传递所需的剂量下是无毒的。我的研究策略是将氨基酸化学连接到纳米金刚石上，以促进它们与核酸的相互作用。该计划的新奇在于与生物活性分子，氨基酸缀合的纳米颗粒的合成和表征方法，以及阐明其与哺乳动物细胞，器官和全身的有益和有害相互作用。本论文的短期研究目标是在前期工作的基础上，设计自组装的脂质基和金刚石基纳米粒子。为了实现我们的目标，我们用氨基酸修饰纳米金刚石的表面，从而得到适合结合核酸的功能化纳米颗粒。我们将评估这些功能化纳米颗粒将核酸递送到模型细胞中的能力。将监测各种官能化纳米金刚石的生物分布。将建立结构/架构-递送效率/毒性之间的关系。这将导致选择具有用于将核酸递送到细胞中的最佳性质（改善的结合性质、细胞摄取和高活性，而没有或具有最小的毒性，以及在生物环境中的稳定性）的候选载体。为了研究递送系统的结构，将使用高分辨率显微镜、流式细胞术和基于同步加速器的技术。将这些纳米颗粒的表面性质与递送效率相关联将允许绘制通用和载体特异性递送机制。从长远来看，我们将探索如何将纳米金刚石颗粒的化学功能化的基本知识扩展到其他固体-核心和软纳米颗粒，建立核酸的通用递送原则。生物分布与表面性质的相关性将被用作预测具有类似表面性质的其他人造纳米颗粒的生物命运的模型。这项研究计划将推进知识的使用和安全性的功能化金刚石-核心纳米粒子作为核酸输送系统。优化这些传递系统的特性将有利于纳米技术（全球市场为12亿美元），最终导致人类和动物的基因治疗。