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Image-based Identification of Chemical Compounds Capable of Trapping FOXO in the Cell Nucleus

Susana Machado1, Catarina Raposo1, Bibiana I Ferreira1,2

  • 1Centre for Biomedical Research (CBMR), University of Algarve, Faro, Portugal.

Methods in Molecular Biology (Clifton, N.J.)
|November 11, 2018
PubMed

Insights

We developed a high-throughput screening assay to identify drugs that trap tumor suppressor proteins, Forkhead box O (FOXO) factors, in the nucleus. This approach could lead to new cancer and age-related disease treatments.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Pharmacology

Background:

  • Forkhead box O (FOXO) proteins are tumor suppressors frequently inactivated in human cancers.
  • Genetic variations in FOXO3a are linked to human longevity.
  • FOXO inactivation typically involves cytoplasmic mislocalization due to posttranslational modifications.

Purpose of the Study:

  • To develop a cell-based imaging assay for screening chemical agents that inhibit nuclear export.
  • To identify compounds that promote nuclear localization of FOXO proteins for therapeutic applications.
  • To establish a high-throughput screening method for monitoring nuclear export.

Main Methods:

  • Development of a cell-based imaging assay using fluorescent-tagged reporter proteins.
  • Screening for chemical agents that inhibit nuclear export of proteins containing a nuclear export sequence.
  • Monitoring changes in fluorescent signal localization (cytoplasmic vs. nuclear).

Main Results:

  • The assay successfully distinguishes between cytoplasmic and nuclear localization of reporter proteins.
  • Untreated cells show predominantly cytoplasmic localization of fluorescent signals.
  • Inhibitors of nuclear export cause reporter proteins to accumulate in the nucleus.

Conclusions:

  • Pharmacological activation of FOXO proteins by promoting nuclear localization is a promising therapeutic strategy.
  • The developed assay is suitable for high-throughput screening of compound libraries.
  • This approach holds potential for treating cancer and age-related diseases in a personalized medicine context.
Keywords:
CMR1FOXOHigh-content screeningNuclear export

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