High-content image-based screening for small-molecule chaperone amplifiers in heat shock.

Area of Science:

• Cellular Biology
• Biochemistry
• Pharmacology

Background:

• Heat shock proteins (HSPs) are crucial for cellular stress response and preventing protein misfolding diseases.
• Small-molecule HSP amplifiers have demonstrated therapeutic promise in preclinical models.
• HSF1 and HSP70 are key regulators of the cellular stress response, involving nuclear stress granule formation and nucleolar translocation.

Purpose of the Study:

• To develop and validate an image-based, multiparametric assay for screening HSF1/HSP70 amplifiers.
• To identify novel small molecules that enhance HSP expression and confer cellular protection.

Main Methods:

• Developed a high-content screening assay in heat-shocked HeLa cells to monitor HSF1/HSP70 stress granule formation.
• Screened a compound library using the robust assay (Z' = 0.62).
• Evaluated the cytoprotective effects of identified compounds against oxygen-glucose deprivation and rotenone-induced stress.

Main Results:

• The assay successfully identified compounds that amplify HSP70 induction in stressed cells without affecting unstressed cells.
• Screening demonstrated high robustness suitable for large-scale application.
• Selected hit compounds provided significant cytoprotection against various cellular stresses.

Conclusions:

• High-content screening of HSF1/HSP70 amplifiers is a practical approach for discovering therapeutics.
• This strategy holds potential for treating protein misfolding diseases.
• The developed assay is effective for identifying compounds that enhance cellular stress resilience.