A high-throughput small molecule screen identifies synergism between DNA methylation and Aurora kinase pathways for X reactivation

作者：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^*

来源：Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(50): 14366-14371.

DOI：10.1073/pnas.1617597113

• X reactivation
• high-throughput screen
• small molecules
• Aurora kinase
• DNA methyltransferase

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 similar to 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.

• 出版日期2016-12-13
• 单位NIH