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

Document text

Principal Investigator: Teri Wang Odom
Organization: NORTHWESTERN UNIVERSITY
Fiscal Year: 2020
Award: $198,306
Funding agency: National Institute of General Medical Sciences

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.

Terms: <3-D><3-Dimensional><3D><Affect><Behavior><Binding><Biologic Models><Biological><Biological Function><Biological Models><Biological Process><Cell Body><Cell Communication><Cell Communication and Signaling><Cell Components><Cell Interaction><Cell Signaling><Cell Structure><Cell membrane><Cell-to-Cell Interaction><Cells><Cellular Membrane><Cellular Structures><Chemicals><Chemistry><Clinical Trials><Coloring Agents><Complex><Custom><Cytoplasmic Membrane><DNA><Deoxyribonucleic Acid><Diagnostic><Differential Interference Contrast Microscopy><Drug Delivery><Drug Delivery Systems><Dyes><Endosomes><Environment><Eukaryotic Cell><Event><FDA approved><Fluorescence><Functional Imaging><Glioblastoma><Goals><Gold><Grade IV Astrocytic Neoplasm><Grade IV Astrocytic Tumor><Grade IV Astrocytoma><IR/UV/Raman Spectroscopy><Image><Imaging Device><Imaging Instrument><Imaging Tool><Immune response><Immunological response><Intracellular Communication and Signaling><Label><Ligands><Lipid Bilayers><Mediating><Membrane><Membrane Protein Gene><Membrane Proteins><Membrane-Associated Proteins><Microscope><Microscopy><Model System><Molecular><Molecular Interaction><Monitor><Multimodal Imaging><Multiple Partners><Nomarski Interference Contrast Microscopy><Nucleic Acids><Oligo><Oligonucleotides><Optics><Organelles><Particle Size><Physiologic><Physiologic Imaging><Physiological><Plasma Membrane><Play><Position><Positioning Attribute><Process><Property><Public Health><Raman Spectroscopy><Raman Spectrum Analysis><Raman spectrometry><Receptosomes><Role><Rotation><Shapes><Short interfering RNA><Signal Transduction><Signal Transduction Systems><Signaling><Small Interfering RNA><Structure><Structure-Activity Relationship><Surface><Surface Proteins><Therapeutic><Time><Treatment Efficacy><Work><aptamer><base><biological signal transduction><chemical structure function><density><design><designing><experiment><experimental research><experimental study><glioblastoma multiforme><host response><imaging><immunoresponse><in vivo><insight><interest><intervention efficacy><lipid bilayer membrane><live cell image><live cell imaging><live cellular image><live cellular imaging><machine learned algorithm><machine learning algorithm><membrane structure><multi-modal imaging><multi-modality><multi-modality imaging><multimodality><multimodality imaging><nano meter scale><nano meter sized><nano particle><nano particle drug><nano probe><nano scale><nano-sized particle><nanometer scale><nanometer sized><nanoparticle><nanoparticle drug><nanoprobe><nanoscale><nanosized particle><oligos><optic imaging><optical><optical imaging><particle><physiological imaging><plasmalemma><plasmonics><programs><response><siRNA><simulation><social role><spongioblastoma multiforme><structure function relationship><therapeutic efficacy><therapeutically effective><therapy efficacy><three dimensional><tool><trafficking><tumor><uptake>