Quality Control for Single Cell Imaging Analytics Using Endocrine Disruptor-Induced Changes in Estrogen Receptor...

Area of Science:

• Environmental Health Sciences
• Toxicology
• Molecular Biology

Background:

• Endocrine disrupting chemicals (EDCs) are pervasive environmental toxicants linked to human disease.
• EDCs often target nuclear receptors, modulating their activity and levels.
• Single-cell analysis of EDC effects on endogenous receptors has been limited by a lack of quality control metrics.

Purpose of the Study:

• To develop and validate single-cell imaging and informatic workflows for querying EDC effects.
• To assess the utility of single-cell estrogen receptor (ER) distribution as a sensitive and reproducible measure of EDC activity.
• To establish quality control metrics for single-cell analysis of EDCs.

Main Methods:

• High-throughput microscopy and image analytics were used to measure single-cell ER nuclear levels.
• A two-tiered quality control pipeline was developed for single-cell analysis.
• The pipeline was tested against numerous biological replicates and toxicants from regulatory lists.

Main Results:

• Single-cell ER distribution and its changes upon chemical exposure proved to be stable and reproducible features.
• The developed method demonstrated high sensitivity in identifying endocrine disruptor toxicants.
• Novel EDCs active solely on endogenous ER were identified using single-molecule RNA FISH.

Conclusions:

• Single-cell ER distribution analysis is a valuable tool for quality control in high-throughput screening of environmental toxicants.
• This approach enables sensitive and reproducible identification of EDCs.
• Coupled with orthogonal assays, it serves as a robust resource for environmental toxicant screening.