Screen for small molecules increasing the mitochondrial membrane potential.

Area of Science:

• Biochemistry
• Cell Biology
• Pharmacology

Background:

• Mitochondrial function is crucial for cellular energy production and has therapeutic implications.
• Identifying small molecules to modulate mitochondrial respiration is vital for research and drug development.
• Current methods for screening mitochondrial modulators can be inefficient.

Purpose of the Study:

• To develop and validate a high-throughput yeast-based screening assay for identifying compounds that enhance mitochondrial function.
• To test the identified compounds in mammalian cell lines for therapeutic potential.
• To characterize the molecular mechanisms of promising compounds in hepatocytes.

Main Methods:

• Utilized a yeast assay to screen 13,680 compounds for increased mitochondrial membrane potential and adenosine triphosphate (ATP) levels.
• Performed secondary screening of hit compounds in mammalian cell types including myotubes, fibroblasts, PC-12, and HepG2 cells.
• Employed genomic and mitochondrial proteomic analyses to investigate the effects of compounds on hepatocytes.

Main Results:

• Identified 14 hit compounds from the initial yeast screen.
• Two compounds demonstrated increased ATP levels in hepatocytes, and two others increased ATP in fibroblasts.
• Characterized molecular changes in hepatocytes, including alterations in key metabolic and signaling factors like PGC-1α and β-Klotho.

Conclusions:

• The yeast-based screening approach is effective for high-throughput identification of mitochondrial function enhancers.
• The identified compounds show potential for modulating cellular energy metabolism in mammalian cells.
• Further investigation into these compounds could lead to novel therapeutic strategies for mitochondrial dysfunction.