Title: A high-throughput small molecule screen identifies synergism between DNA methylation and Aurora kinase pathways for X reactivation.
Authors: Lessing, Derek; Dial, Thomas O; Wei, Chunyao; Payer, Bernhard; Carrette, Lieselot L G; Kesner, Barry; Szanto, Attila; Jadhav, Ajit; Maloney, David J; Simeonov, Anton; Theriault, Jimmy; Hasaka, Thomas; Bedalov, Antonio; Bartolomei, Marisa S; Lee, Jeannie T
Published In Proc Natl Acad Sci U S A, (2016 12 13)
Abstract: X-chromosome inactivation is a mechanism of dosage compensation in which one of the two X chromosomes in female mammals is transcriptionally silenced. Once established, silencing of the inactive X (Xi) is robust and difficult to reverse pharmacologically. However, the Xi is a reservoir of >1,000 functional genes that could be potentially tapped to treat X-linked disease. To identify compounds that could reactivate the Xi, here we screened ∼367,000 small molecules in an automated high-content screen using an Xi-linked GFP reporter in mouse fibroblasts. Given the robust nature of silencing, we sensitized the screen by "priming" cells with the DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (5azadC). Compounds that elicited GFP activity include VX680, MLN8237, and 5azadC, which are known to target the Aurora kinase and DNA methylation pathways. We demonstrate that the combinations of VX680 and 5azadC, as well as MLN8237 and 5azadC, synergistically up-regulate genes on the Xi. Thus, our work identifies a synergism between the DNA methylation and Aurora kinase pathways as being one of interest for possible pharmacological reactivation of the Xi.
PubMed ID: 28182563
MeSH Terms: Animals; Aurora Kinase A/antagonists & inhibitors; Aurora Kinase A/genetics; Aurora Kinase B/antagonists & inhibitors; Aurora Kinase B/genetics; Aurora Kinases/antagonists & inhibitors*; Aurora Kinases/genetics; Azacitidine/administration & dosage; Azacitidine/analogs & derivatives; Azepines/administration & dosage; Cell Line; DNA Methylation/drug effects*; Drug Evaluation, Preclinical; Drug Synergism; Female; Gene Knockdown Techniques; Genes, X-Linked; Green Fluorescent Proteins/genetics; Green Fluorescent Proteins/metabolism; High-Throughput Screening Assays; Mice; Mice, Transgenic; Piperazines/administration & dosage; Pyrimidines/administration & dosage; X Chromosome Inactivation/drug effects*; X Chromosome/drug effects; X Chromosome/genetics