Engineering functionalized diamond nanoparticles for intracellular delivery of nucleic acids

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。在过去的二十年里，基因递送一直是一个重要的研究领域，但主流的化学基因递送纳米材料、脂质和聚合物在其稳定性、效率和毒性方面存在局限性。我的研究项目主要集中在使用固体核纳米颗粒，如纳米金刚石，以克服这些障碍。纳米金刚石凭借均匀的尺寸分布和可用于化学改性的大表面积而具有优于脂质和聚合物纳米颗粒的优点。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方法。*短期研究目标建立在我以前的工作基础上：设计自组装的基于脂质和基于金刚石的纳米颗粒。为了实现我们的目标，我们将化学连接纳米金刚石表面上的氨基酸，从而产生具有适合结合核酸的表面性质的功能化纳米颗粒。我们将评估这些功能化纳米颗粒将结合的核酸递送到模型哺乳动物细胞中的能力。将研究细胞摄取途径和潜在毒性机制。将建立结构/架构-递送效率/毒性之间的关系。这将导致鉴定具有用于将核酸递送到细胞中的最佳特性（改进的结合特性、稳定性和细胞摄取，而没有毒性或毒性最小）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，我们将探索纳米金刚石颗粒的化学功能化的基础知识是否可以扩展到其他固体核和软纳米颗粒，建立核酸的通用递送原则。这项研究计划将推进功能化金刚石核纳米粒子作为核酸递送系统的使用和安全性的知识。这些递送系统特征的优化将有利于纳米技术药物递送（全球市场规模为12亿美元，未来十年预计增长率为49%），最终导致人类和动物的基因治疗。了解纳米金刚石与核酸的相互作用也可以转化为生物传感器应用。

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.******