Screening and identification of small molecule compounds perturbing mitosis using time-dependent cellular response...

Area of Science:

• Cell Biology
• Pharmacology
• Biophysics

Background:

• Cellular processes like mitosis are dynamic and time-dependent.
• Understanding how compounds perturb these processes requires temporal measurement.
• Existing methods may lack the resolution to capture nuanced temporal responses.

Purpose of the Study:

• To develop and validate a time-resolved impedance-based assay for cellular response profiling.
• To identify novel antimitotic compounds using this profiling approach.
• To characterize the temporal signatures of cellular responses to antimitotic perturbations.

Main Methods:

• Utilized impedance-based time-dependent cell response profiling (TCRP) to monitor cellular dynamics.
• Applied TCRP to measure responses to known antimitotic compounds and siRNAs.
• Screened a library of 119,595 compounds using TCRP signatures and clustering analysis.
• Validated hit compounds through independent antimitotic assays.

Main Results:

• Small molecule perturbation of mitosis generated unique and identifiable TCRP signatures.
• TCRP-based screening identified novel antimitotic compounds with high accuracy (113/117 validated).
• The approach identified potent agents interfering with tubulin polymerization and other mitotic mechanisms.

Conclusions:

• TCRP is a robust and predictive phenotypic profiling tool for studying cellular pathways.
• The time-resolved nature of TCRP offers a valuable complement to existing multidimensional profiling methods.
• This approach enables practical, unbiased screening for novel therapeutic agents targeting cell cycle regulation.