Novel Self Assembly of siRNA for Efficient and Safe Delivery

新型 siRNA 自组装技术可实现高效、安全的递送

• 批准号: 0933966
• 负责人: Huixin He
• 金额: $ 30万
• 依托单位: Rutgers University Newark
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0933966&HistoricalAwards=false
• 关键词: Novel; Self; Assembly; siRNA; Efficient

0933966HeThis proposal describes an integrated approach for interdisciplinary research, education, and outreach in nanotechnology and biomedical engineering. The objective of the research is to develop an innovative and nontoxic delivery platform that will enable cell specific delivery of small interference RNAs (siRNAs) to silence their targeted oncogenes both in vitro and in vivo. The other objective of this research is to seek an extensive understanding of the fundamental physicochemical characteristics, especially nanomechanical properties, of siRNA nanoparticles with their biological performance. To reach these goals, the proposal has three associated specific aims: (1) To develop a novel approach for assembly and delivery of siRNA without toxicity using labile Au nanoparticles modified with several low generation dendrimers. (2) To engineer the surface of siRNA nanoparticles for target delivery, and to study the mechanical properties of the individual siRNA nanoparticles (both with and without Au nanoparticles encapsulated, and both engineered and non-engineered). (3) To determine structure/property-biodistribution, biological potency, and toxicity relationships of the siRNA nanoparticles in vitro and in vivo. Intellectual Merit: A novel siRNA assembly approach where Au nanopartices (Au NPs) will be used to dramatically enhance non toxic, low-generation dendrimers to efficiently condense siRNA to discrete nanoparticles. However, the Au NPs can be controlled "in" or "out" of the final siRNA complexes, which is the key difference from previous reports. The potential toxic problem accompanied with the Au NPs will be solved by selectively removing the Au NPs before the siRNA complexes are delivered. In addition, to satisfy the requirements for in vivo targeted delivery of siRNA through a systemic route, the formed siRNA nanoparticles will be engineered by a layer-by-layer modular approach to enable them for spatially- and temporally- controlled release in specific sub-cellular compartments. These properties will add additional therapeutic activities and further decrease the side effects of RNAi-based therapy.In addition to the physicochemical properties, the nanomechanical properties of the individual siRNA nanoparticles from various formulations will be studied by single force microscopy. By combining the biological investigation of these siRNA nanoparticles, this proposal will link, for the first time, the physicochemical properties and the nanomechanical properties of the siRNA nanoparticles with their cellular internalization, circulation, biodistribution, and therefore, their targeting and therapeutic efficacy during in vitro and in vivo systemic delivery. The improved understanding of these relationships will lead to future design and development of new materials and strategies for efficient and safe delivery of siRNA, and therefore help to realizing its full therapeutic potentials. Successful completion of this research will also provide critical understanding how we can utilize information obtained from various multifunctional nanomedicine platforms which are constructed by engineered inorganic nanocarriers (relatively hard) to guide the development of efficient organic nanocarriers (relatively soft) and vice versa. Broader Impacts: The proposed investigations are fundamentally and practically important for efficient and safe siRNA delivery. The proposal focuses on design multifunctional siRNA nanoparticles capable of cell specific delivery and silencing of gene expression of EZH2 genes for breast cancer therapy. Given the widespread applications of siRNA in numerous fundamental and therapeutic applications, the knowledge gained from this project will have far reaching scientific and economic impacts on pharmaceutical and biotechnology industry and health care. The educational plan will bring nanoscience tools and concepts to a wide range of students on two campus of Rutgers known as the most diverse in the nation. The inherently interdisciplinary nature of this research will produce students with exceptional training in nanotechnology, biomedical engineering, molecular biology, and drug delivery. Research activities designed for undergraduates and high school students will promote more gifted minority students into the nanoscience ranks. Extensive outreach to the Newark area, a minority-dominated region, will raise the public awareness of the impact of nanoscience and nanotechnology.

0933966 He该提案描述了纳米技术和生物医学工程的跨学科研究，教育和推广的综合方法。该研究的目的是开发一种创新的无毒递送平台，使细胞特异性递送小干扰RNA（siRNA），以在体外和体内沉默其靶向癌基因。本研究的另一个目的是寻求对siRNA纳米颗粒的基本物理化学特性，特别是纳米力学特性及其生物学性能的广泛理解。为了达到这些目标，该提案有三个相关的具体目标：（1）开发一种新的方法，用于组装和交付siRNA无毒性使用不稳定的Au纳米粒子修饰的几个低代树枝状聚合物。(2)设计siRNA纳米颗粒的表面以用于靶向递送，并研究单个siRNA纳米颗粒的机械性质（有和没有封装Au纳米颗粒，以及工程化和非工程化）。(3)确定siRNA纳米颗粒的结构/性质-生物分布、生物学效力和毒性关系。智力优势：一种新的siRNA组装方法，其中Au纳米颗粒（Au NPs）将用于显著增强无毒、低代树枝状聚合物以有效地将siRNA浓缩到离散的纳米颗粒。然而，Au NPs可以被控制在最终siRNA复合物的“内”或“外”，这是与先前报道的关键区别。伴随Au NP的潜在毒性问题将通过在递送siRNA复合物之前选择性地去除Au NP来解决。此外，为了满足通过全身途径体内靶向递送siRNA的要求，所形成的siRNA纳米颗粒将通过逐层模块化方法进行工程化，以使它们能够在特定的亚细胞区室中进行空间和时间控制释放。这些特性将增加额外的治疗活性，并进一步减少基于RNAi的治疗的副作用。除了物理化学性质，来自各种配方的单个siRNA纳米颗粒的纳米机械性质将通过单力显微镜进行研究。通过结合这些siRNA纳米颗粒的生物学研究，该提案将首次将siRNA纳米颗粒的物理化学性质和纳米机械性质与其细胞内化、循环、生物分布联系起来，从而将其在体外和体内全身递送期间的靶向和治疗功效联系起来。对这些关系的进一步理解将导致未来设计和开发新材料和策略，以有效和安全地递送siRNA，从而有助于实现其全部治疗潜力。这项研究的成功完成还将提供关键的理解，我们如何利用从各种多功能纳米医学平台获得的信息，这些平台由工程无机纳米载体（相对较硬）构建，以指导有效的有机纳米载体（相对较软）的开发，反之亦然。更广泛的影响：所提出的研究对于有效和安全的siRNA递送具有根本性和实际重要性。该提案的重点是设计能够细胞特异性递送和沉默EZH 2基因表达的多功能siRNA纳米颗粒，用于乳腺癌治疗。鉴于siRNA在许多基础和治疗应用中的广泛应用，从该项目中获得的知识将对制药和生物技术行业以及医疗保健产生深远的科学和经济影响。该教育计划将为罗格斯大学两个校园的广泛学生带来纳米科学工具和概念，这两个校园被称为全国最多样化的校园。这项研究固有的跨学科性质将产生学生在纳米技术，生物医学工程，分子生物学和药物输送的特殊培训。为本科生和高中生设计的研究活动将促进更多有天赋的少数民族学生进入纳米科学行列。广泛推广到纽瓦克地区，一个少数民族占主导地位的地区，将提高公众对纳米科学和纳米技术的影响的认识。