Shape Control and Transport Properties of DNA-Copolymer Micelles

 描述：模仿病毒颗粒大小和形状的胶束纳米颗粒作为DNA输送载体具有吸引力，因为它们改善了胶体稳定性和运输性能，能够逃避免疫清除，以及高有效载荷包装能力。此外，纳米颗粒的形状已被明确地确定为决定其传输性能和传递效率的重要因素。然而，还没有可用的纳米颗粒合成方法来包装质粒DNA有效载荷，同时允许足够的颗粒大小和形状控制。最近，我们已经证明，在可控的组装条件下，通过将质粒DNA与工程嵌段或接枝的聚阳离子和聚乙二醇共聚物络合，可以实现对DNA胶束的明显形状控制和调节。在这项拟议的研究中，我们将开发一个包括并行和集成的实验和计算策略的协同研究计划，以(1)开发和了解DNA胶束组装的新方法，以实现形状可控、高度均匀的合成，并具有高稳定性；(2)揭示形状依赖的纳米颗粒在体外和体内生理介质中的扩散和传输特性；以及(3)通过形状可控的DNA胶束展示治疗载体的传递效率以及它们的成像和治疗效果。这项拟议的研究将DNA纳米颗粒组装、基于微流体的单颗粒分析/荧光相关光谱、活体成像、癌症治疗和计算机模拟方面的独特专业知识结合在一起，以解决DNA纳米治疗技术的工程和交付方面的关键知识缺口。这不仅将为合成形状可控的DNA胶束提供一种新的、可推广的方法，而且还将从机理上理解纳米粒子的形状相关传输特性。此外，我们的实验和计算方法的集成性质建立了一个新的范式，将极大地加速新的DNA纳米颗粒系统的发现和开发，以实现高效的基因药物输送。

 DESCRIPTION: Micellar nanoparticles that mimic the size and shape of viral particles are attractive as a DNA delivery vehicle because of their improved colloidal stability and transport properties, ability to evade immune clearance, and high payload packaging capacity. Moreover, nanoparticle shape has explicitly been identified as an important factor determining their transport properties and delivery efficiency. However, there is no available nanoparticle synthesis method for packaging plasmid DNA payloads while allowing sufficient control over particle size and shape. Recently, we have shown that distinct shape control and tuning for DNA micelles can be achieved through complexation of plasmid DNA with engineered block or graft copolymers of polycation and poly (ethylene glycol) under controlled assembly conditions. In this proposed study, we will develop a synergistic research program comprising parallel and integrated experimental and computational strategies to (1) develop and understand new methods for DNA micelle assembly that permit scalable, high-uniformity synthesis with shape control and high stability; (2) reveal shape-dependent nanoparticle diffusion and transport properties in physiologically media in vitro and in vivo; and (3) demonstrate the delivery efficiency of a theranostic vector by shape-controlled DNA micelles and their imaging and therapeutic efficacy using mouse models of human metastatic cancers. The proposed study brings together a unique combination of expertise in DNA nanoparticle assembly, microfluidics-based single-particle analysis/fluorescence correlation spectroscopy, in vivo imaging, cancer theranostics, and computer simulations to address a crucial knowledge gap in the engineering and delivery of DNA nano-therapeutics. It will not only offer a new, generalizable method for synthesizing shape-controlled DNA micelles, but also provide a mechanistic understanding of shape- dependent transport properties of nanoparticles. Moreover, the integrated nature of our experimental and computational approach establishes a new paradigm that will greatly accelerate the discovery and development of new DNA nanoparticle systems for efficient gene medicine delivery.