HIGH CONTENT ANALYSIS TO IDENTIFY MODULATORS OF HUNTINGTIN N17 PHOSPHORYLATION

Abstract

The N17 domain in the huntingtin protein is an important modulator of mutant huntingtin toxicity in Huntington’s Disease (HD). Notably, this domain is hypophosphorylated in HD, and promoting its phosphorylation is protective in HD models. I conducted high content analysis of a library of 1163 compounds to identify modulators of phospho-N17 in order to gain mechanistic insight into the pathways which affect N17 phosphorylation. I found that the pharmaceutical compound Pranlukast promoted N17 phosphorylation by inhibiting the NFκB signaling pathway, which was also shown to modulate N17 phosphorylation in our lab’s previous work. Throughout the course of this project, I developed a novel method for screening moderately sized compound libraries that is inexpensive, unbiased, sensitive, and accurate. This new high content analysis procedure, known as the “Gladiator” screening method, can be used as a framework to screen compounds in small laboratory.