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High-Throughput Screening for Drugs that Modulate Intermediate Filament Proteins
Jingyuan Sun1, Vincent E Groppi2, Honglian Gui3
1Department of Molecular & Integrative Physiology, University of Michigan, Ann Arbor, Michigan, USA; Department of Medicine, University of Michigan, Ann Arbor, Michigan, USA; VA Ann Arbor Healthcare System, Ann Arbor, Michigan, USA; Hepatology Unit, Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, PR China; Department of Radiation Oncology, Nanfang Hospital, Southern Medical University, Guangzhou, PR China.
Researchers developed a high-throughput drug screening method to identify compounds that can normalize disrupted intermediate filament (IF) proteins. This approach offers a potential therapeutic strategy for numerous intermediate filament-related diseases (IF-pathies).
Area of Science:
- Cell Biology
- Biochemistry
- Genetics
Background:
- Intermediate filament (IF) proteins exhibit complex tissue-specific expression.
- Mutations in IF genes lead to over 80 human diseases (IF-pathies), often caused by disrupted IF networks.
- Current therapies address IF-pathy complications, not the underlying protein defects.
Purpose of the Study:
- To develop a high-throughput screening method for identifying drugs that normalize aberrant IF proteins.
- To establish a strategy applicable to various IF proteins and filament-disrupting mutations.
- To facilitate the discovery of novel therapeutics for IF-pathies.
Main Methods:
- Utilized lentiviral transduction to express green fluorescent protein-tagged keratin 18 (K18) R90C mutant in A549 cells.
- Developed a readout assay to detect compounds that restore wild-type-like filamentous IF distribution from a dot-like pattern.
- Screened thousands of compounds for their ability to normalize mutant IF proteins.
Main Results:
- Identified drug "hits" capable of converting the dot-like keratin filament distribution caused by the R90C mutation to a filamentous array.
- Demonstrated the adaptability of the screening strategy for different IF proteins with filament-disrupting mutations.
- Established methods for validating drug hits in cell culture and in vivo models.
Conclusions:
- The high-throughput screening approach can identify compounds that normalize disrupted IF proteins.
- This method holds promise for developing targeted therapies for a wide range of IF-pathies.
- The ultimate goal is to find safe and effective drugs for clinical testing in patients with IF-related diseases.
