Nanomedicine:Supra-molecular cellular compartments (RMI)

纳米医学：超分子细胞区室（RMI）

• 批准号: 6930773
• 负责人: Edward N Pugh
• 金额: $ 5.35万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6930773
• 关键词: cell component structure /function; confocal scanning microscopy; fluorescence microscopy; molecular assembly /self assembly; nanomedicine; nanotechnology; tissue /cell culture

The long-term goals of the University of Pennsylvania Nanomedicine Development Center (NDC) are to characterize quantitatively at the molecular scale the components and functional organization of selected supra-molecular cellular compartments (SMCCs), and to develop the nanoscale tools required to repair or replace these components in diseased cells. Many developmental and acquired diseases are caused by deficits in assembly or repair of specialized SMCCs. The molecular details of how cells accomplish the programmed self-assembly of complex compartments, such as photoreceptor outer segments, primary cilia in the renal tubules, the nerve growth cone, and the myofibril, have tremendous health-care relevance and intrinsic nanotechnological interest. We will combine state-of-the-art molecular biologic techniques with both existing and new nanotechniques to investigate the specific requirements of the individual SMCC components, the order of their assembly, their intramolecular contacts and interactions, and the crucial involvement of accessory proteins, such as chaperones and molecular motors. Improved understanding of these details will have two important outcomes: Firstly, replacement of defective components will be more readily targeted in disease processes, such as retinal degenerations, polycystic kidney disease and cardiomyopathy. Secondly, full understanding of the assembly requirements will allow design of practical, artificial nanotechnological devices, such as sensors, actuators and delivery vehicles, using the same principles of specificity and self-assembly. We will refine and develop several nanomedical tools for the proposed research. For example, we will perfect the two-photon confocal microscopy techniques required to quantitate individual SMCC components and follow their movements in real time within living cells. We will develop the use of synthetic polymersome nanoparticles to deliver fluorescently labeled SMCC proteins to living cells on the microscope stage for these experiments. We will use total internal reflection epifluorescence microscopy (TIRF) techniques to monitor cell-free assembly of SMCC component nanomachines. The use of modified polymersome and viral vector nanoparticles will be developed for delivery of essential SMCC components to cells in culture and in vivo. In addition, we will investigate new techniques for measurement of intracellular assembly of SMCC components at high resolution using nanotubes as optical sensors. These research tools will be useful for the study of multiple SMCCs, and broadly applicable to nanomedicine research in general.

宾夕法尼亚大学纳米医学开发中心（NDC）的长期目标是在分子尺度上定量表征所选超分子细胞隔室（SMCC）的组件和功能组织，并开发用于修复或替代这些分离细胞中所需的纳米级工具。许多发育和获得的疾病是由专业SMCC的组装或修复缺陷引起的。细胞如何完成复杂区室的编程自组装的分子细节，例如感光体外部段，肾小管中的原发性纤毛，神经生长锥和肌原纤维具有巨大的医疗保健相关性和内在的纳米技术兴趣。我们将结合最先进的分子 具有现有和新纳米技术的生物学技术，以研究单个SMCC组件的特定要求，组装顺序，分子内接触和相互作用以及辅助蛋白（例如伴侣蛋白和分子运动）的关键参与。改善了对这些细节的理解 将有两个重要的结果：首先，替换有缺陷的成分将更容易针对疾病过程，例如视网膜退化，多囊性肾脏疾病和心肌病。其次，对组装要求的充分理解将允许使用相同的特殊性和自组装原理设计实用的人工纳米技术设备，例如传感器，执行器和送货工具。我们将完善并开发几种纳米医学工具进行拟议的研究。例如，我们将完善定量单个SMCC组件并实时跟随活细胞内的两光子共聚焦显微镜技术。我们将开发使用合成多颗粒纳米颗粒的使用，以在显微镜阶段将荧光标记的SMCC蛋白标记为活细胞进行这些实验。我们将使用总内部反射荧光显微镜（TIRF）技术来监测SMCC组件纳米机器的无细胞组装。将开发使用改性多聚合物体和病毒载体纳米颗粒以提供 培养和体内细胞的必需SMCC成分。此外，我们将研究使用纳米管作为光学传感器的高分辨率测量SMCC组件细胞内组件的新技术。这些研究工具将用于多个SMCC的研究，并且通常适用于纳米医学研究。