Physiologically Relevant Estrogen Receptor Alpha Pathway Reporters for Single-Cell Imaging-Based Carcinogenic Hazard...

Area of Science:

• Endocrinology
• Molecular Biology
• Toxicology

Background:

• Estrogen receptor alpha (ERα) is a nuclear hormone receptor regulating cell growth and proliferation.
• Disruption of ERα pathways by estrogenic compounds can lead to adverse outcomes, including tumor formation.
• Quantifying key events in ERα-mediated adverse outcomes is crucial for distinguishing toxic effects from normal physiological responses.

Purpose of the Study:

• To develop novel fluorescent reporter cell lines for monitoring ERα-induced proliferation.
• To establish tools for quantifying the dynamics of ERα pathway activation at the single-cell level.
• To assess the potential carcinogenic activity of estrogenic compounds.

Main Methods:

• Established MCF7 reporter cell lines using bacterial artificial chromosome-based tagging of three ERα target genes: GREB1, PGR, and TFF1.
• Characterized the reporter cell lines' response to 17β-estradiol and other reference estrogenic compounds.
• Evaluated the activation dynamics and sensitivities of the reporter cell lines.

Main Results:

• The developed GFP reporter cell lines exhibited distinct activation dynamics upon exposure to 17β-estradiol.
• Reporter cell lines demonstrated specific activation patterns in response to various estrogenic compounds with potencies comparable to OECD test methods.
• The cell lines allow for monitoring spatial and temporal dynamics of ERα pathway activation.

Conclusions:

• Fluorescent reporter cell lines provide mechanistic insights into ERα pathway activation.
• These tools facilitate a detailed assessment of the carcinogenic potential of estrogenic compounds.
• The study offers a novel approach for evaluating endocrine disruptors and their impact on human health.