High-Throughput Screening for Drugs that Modulate Intermediate Filament Proteins

摘要: Intermediate filament (IF) proteins have unique and complex cell 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 at conserved residues that result in a disrupted network. A critical need for entire IF-pathy field is identification of drugs can ameliorate cure these diseases, particularly since all current therapies target complication, such as diabetes cardiovascular disease, rather mutant protein gene. We describe high-throughput approach identify normalize proteins. This utilizes transduction lentivirus expresses green fluorescent protein-tagged keratin 18 (K18) R90C A549 cells. The readout drug "hits" convert dot-like distribution, mutation, wild-type-like filamentous array. similar strategy be used screen thousands compounds utilized practically any filament-disrupting could therefore potentially many IF-pathies. "Hits" interest require validation culture then using vivo experimental models. Approaches study mechanism normalization by potential are also described. ultimate goal this screening effective safe tested clinical efficacy patients.