Nano Calcium Phosphate for Gene Delivery

用于基因传递的纳米磷酸钙

• 批准号: 7098876
• 负责人: PRASHANT NAGESH KUMTA
• 金额: $ 3.47万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-20 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7098876
• 关键词: DNA; X ray crystallography; calcium phosphate; collagen; digital imaging; extracellular matrix; fibrin; gene delivery system; gene expression; gene therapy; histochemistry /cytochemistry; infrared spectrometry; interferometry; laboratory rat; plasmids; polymerase chain reaction; tissue /cell culture; transfection; transmission electron microscopy

DESCRIPTION (provided by applicant): Gene delivery using non-viral (plasmid) techniques are very desirable due to their economic, convenience, ease of manufacturing, cost-effectiveness, and safe characteristics. The techniques are currently limited by low transfection efficiencies. To address this limiting factor, our research focuses on developing nano-sized calcium phosphate (CAP) particles called "NanoCaPs" as a novel delivery agent for plasmid DNA transfection. The reduced crystallite dimensions of nano-sized materials, compared to conventionally synthesized micron sized CaP will result in increased surface-active DNA binding sites and enhanced reactivity, thus improving transfection efficiencies. Furthermore, the NanoCaPs are biocompatible and they could be used alone or incorporated in many synthetic or natural polymers to deliver genes in a sustained manner thus providing an elegant plasmid gene delivery system. The NanoCaPs have the potential to provide nucleic-acid based therapeutics that closely resembles traditional pharmaceuticals and gene delivery for tissue engineering. The overall objective of this grant application is to engineer a safe and versatile plasmid gene delivery system for tissue engineering applications. Our goal is to demonstrate that the synthesized NanoCaPs incorporated in a natural (fibrin, collagen etc.) or synthetic (PLA, PLGA, PCL, polyurethane etc.) biomimetic extracellular matrix (bECM) will enhance both the in vitro and in vivo transfection efficiency of plasmid DNA (pDNA) by increasing the uptake and expression of marker genes (Luciferase and/or LacZ). This will be achieved using quantitative digital imaging methods. Our preliminary in vitro data is in excellent agreement with our objective to design and develop an efficient plasmid gene therapy that could be utilized with or without biodegradable polymers for gene replacement therapy and tissue engineering. The present study will provide a concrete foundation for conducting further applied and basic science research related to plasmid gene therapy. The specific aims have been formulated to provide solutions to fundamental questions related to in vitro and in vivo pDNA transfection efficiency and the application of the pDNA/NanoCaPs/polymer in bone tissue engineering thus providing key information that is currently not available. The proposed research will enable the generation of novel carriers for plasmid gene delivery, the benefits of which will be seen in both hard and soft tissue engineering applications.

描述（由申请人提供）：使用非病毒（质粒）技术的基因递送由于其经济、便利、易于制造、成本效益和安全的特性而非常受欢迎。 该技术目前受到转染效率低的限制。 为了解决这一限制因素，我们的研究重点是开发称为“NanoCaPs”的纳米级磷酸钙 (CAP) 颗粒，作为质粒 DNA 转染的新型递送剂。 与传统合成的微米级 CaP 相比，纳米级材料的微晶尺寸减小，从而增加了表面活性 DNA 结合位点并增强了反应性，从而提高了转染效率。 此外，NanoCaP 具有生物相容性，它们可以单独使用或掺入许多合成或天然聚合物中，以持续的方式传递基因，从而提供优雅的质粒基因传递系统。 NanoCaP 有潜力提供基于核酸的疗法，与传统药物和组织工程基因传递非常相似。 该拨款申请的总体目标是为组织工程应用设计一个安全且多功能的质粒基因传递系统。 我们的目标是证明合成的 NanoCaP 掺入天然（纤维蛋白、胶原蛋白等）或合成（PLA、PLGA、PCL、聚氨酯等）仿生细胞外基质 (bECM) 中将提高体外和体内转染效率通过增加标记基因（荧光素酶和/或 LacZ）的摄取和表达来修饰质粒 DNA (pDNA)。 这将通过定量数字成像方法来实现。 我们的初步体外数据与我们设计和开发有效的质粒基因疗法的目标非常一致，该疗法可以与或不与可生物降解的聚合物一起用于基因替代疗法和组织工程。 本研究将为进一步开展质粒基因治疗相关的应用和基础科学研究提供坚实的基础。 制定的具体目标是为体外和体内 pDNA 转染效率以及 pDNA/NanoCaPs/聚合物在骨组织工程中的应用相关的基本问题提供解决方案，从而提供目前无法获得的关键信息。 拟议的研究将能够产生用于质粒基因传递的新型载体，其好处将在硬组织工程和软组织工程应用中看到。