In vivo imaging of specific drug-target binding at subcellular resolution
J M Dubach1, C Vinegoni1, R Mazitschek2
11] Center for System Biology, Massachusetts General Hospital and Harvard Medical School, Richard B. Simches Research Center, 185 Cambridge Street, Boston, Massachusetts 02114, USA [2].
Nature Communications
|May 29, 2014
Summary
We developed a new live-cell imaging technique to directly measure drug-target binding in real time. This method maps drug distribution and engagement at subcellular resolution, offering a deeper understanding of drug action in vivo.
Area of Science:
- Pharmacology and live-cell imaging
- Drug discovery and development
- Molecular biology and biophysics
Background:
- Current methods for measuring drug-target interactions in live cells are often indirect, static, or require cell lysis, limiting our understanding of drug action.
- Existing techniques provide incomplete insights into the dynamic spatial and temporal distribution of drugs and their binding status within cells.
- A need exists for a direct, real-time method to visualize and quantify drug-target engagement at subcellular resolution.
Purpose of the Study:
- To develop and validate a novel multiphoton fluorescence anisotropy microscopy technique for real-time measurement of drug-target binding in live cells.
- To enable high-resolution spatial and temporal mapping of both bound and unbound drug distribution within cells.
- To demonstrate the general applicability of the technique using a fluorescently labeled drug and assess its utility in vivo.
Main Methods:
- Utilized multiphoton fluorescence anisotropy microscopy for live-cell imaging.
- Developed a method for real-time, subcellular resolution mapping of drug-target interactions.
- Applied the technique to visualize the intracellular target engagement of Olaparib, a poly(ADP-ribose) polymerase inhibitor, in live cells and in vivo.
Main Results:
- Successfully measured and mapped real-time drug-target binding at subcellular resolution.
- Demonstrated high-resolution spatial and temporal mapping of bound and unbound drug distribution.
- Visualized Olaparib's intracellular target engagement in live cells and within a tumor in vivo.
Conclusions:
- The developed multiphoton fluorescence anisotropy microscopy technique provides a generalizable approach for directly measuring drug-target binding in vivo.
- This method offers a powerful new tool for enhancing the understanding of drug activity and action at the molecular level.
- The technique has significant potential for drug discovery and development by enabling direct visualization of drug engagement.
