Continuous Probing of Nanoconstruct-Cell Interactions at Biologically Relevant Time Scales

在生物学相关时间尺度上连续探测纳米结构-细胞相互作用

• 批准号: 10475700
• 负责人: Teri Wang Odom
• 金额: $ 19.73万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-27 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10475700
• 关键词: 3-Dimensional; Affect; Behavior; Binding; Biological; Biological Models; Biological Process; Cell Communication; Cell membrane; Cells; Cellular Membrane; Cellular Structures; Chemicals; Chemistry; Clinical Trials; Complex; Custom; DNA; Diagnostic; Drug Delivery Systems; Dyes; Endosomes; Environment; Eukaryotic Cell; Event; FDA approved; Fluorescence; Functional Imaging; Glioblastoma; Goals; Gold; Image; Imaging Device; Immune response; Label; Ligands; Lipid Bilayers; Mediating; Membrane; Membrane Proteins; Microscope; Microscopy; Molecular; Monitor; Multimodal Imaging; Multiple Partners; Nomarski Interference Contrast Microscopy; Nucleic Acids; Oligonucleotides; Optics; Organelles; Particle Size; Physiological; Play; Positioning Attribute; Process; Property; Public Health; Raman Spectrum Analysis; Role; Rotation; Shapes; Signal Transduction; Small Interfering RNA; Structure; Structure-Activity Relationship; Surface; Therapeutic; Time; Treatment Efficacy; Work; aptamer; base; density; design; experimental study; in vivo; insight; interest; live cell imaging; machine learning algorithm; multimodality; nanoparticle; nanoparticle drug; nanoprobe; nanoscale; optical imaging; particle; plasmonics; programs; response; simulation; tool; trafficking; tumor; uptake

PROJECT SUMMARY This proposal aims to develop live-cell, multi-channel imaging tools that can visualize—continuously, and in real time—nanoparticle interactions with cellular components. We will focus on several different time windows, where early time periods will monitor nanoparticle-cell membrane binding and uptake and later times will track endosomal accumulation and escape. The ability to resolve temporally and spatially how particle size/shape and ligand density affects interactions in 3D is critical for determining structure-activity-relationships and mechanism of action in live cells. To characterize interactions of functional nanoparticles with different cellular structures at physiologically relevant times, we propose to design a multi-channel optical microscope integrated with an opto- splitter and custom live-cell imaging chamber. Simultaneous images can be acquired in different channels of the fluorescence of dye-labeled organelles and dye-labeled ligands on the particles as well as differential interference contrast (DIC) signals of whole cells, cellular components, and nanoparticle cores. Correlation of structural and functional images provides a powerful window into how local nanoconstruct interactions can mediate a biological response. For model systems, we will compare gold nanoconstructs with oligonucleotide ligand shells of both targeting (DNA aptamers) or non-targeting (siRNA) properties. Nanoparticle shape enables a unique handle to probe rotation and orientation of intracellular particle interactions. This work can bridge a gap in understanding the behavior of nanoconstructs intracellularly and how the integrity and presentation of oligonucleotides in ligand shells affects targeting and other processes such as endosomal escape, which is critical to assess therapeutic efficacy.

项目总结