High-Throughput Screening for Drugs that Modulate Intermediate Filament Proteins

Methods in Enzymology
Jingyuan SunM B Omary

Abstract

Intermediate filament (IF) proteins have unique and complex cell and tissue distribution. Importantly, IF gene mutations cause or predispose to more than 80 human tissue-specific diseases (IF-pathies), with the most severe disease phenotypes being due to mutations at conserved residues that result in a disrupted IF network. A critical need for the entire IF-pathy field is the identification of drugs that can ameliorate or cure these diseases, particularly since all current therapies target the IF-pathy complication, such as diabetes or cardiovascular disease, rather than the mutant IF protein or gene. We describe a high-throughput approach to identify drugs that can normalize disrupted IF proteins. This approach utilizes transduction of lentivirus that expresses green fluorescent protein-tagged keratin 18 (K18) R90C in A549 cells. The readout is drug "hits" that convert the dot-like keratin filament distribution, due to the R90C mutation, to a wild-type-like filamentous array. A similar strategy can be used to screen thousands of compounds and can be utilized for practically any IF protein with a filament-disrupting mutation, and could therefore potentially target many IF-pathies. "Hits" of interest require validation in cell c...Continue Reading

Citations

Sep 14, 2017·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Graham F BradyM Bishr Omary
Apr 1, 2017·American Journal of Physiology. Gastrointestinal and Liver Physiology·M Bishr Omary
Aug 5, 2020·Expert Opinion on Therapeutic Patents·Fenju WeiPeng Zhan
Oct 16, 2019·Proceedings of the Japan Academy. Series B, Physical and Biological Sciences·Yuhei NishimuraMasaki Inagaki
Sep 16, 2020·International Journal of Molecular Sciences·Alexander J HinbestChristopher G Bunick
Nov 16, 2020·Current Opinion in Cell Biology·Sherif A EldiranyChristopher G Bunick

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