Small molecules with antioxidative properties are prevalent DUX4 inhibitors within HTS library

Bosnakovski, Darko (2012) Small molecules with antioxidative properties are prevalent DUX4 inhibitors within HTS library. In: FSH Society Facioscapulohumeral Muscular Dystrophy [FSHD].


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Small molecules with antioxidative properties are prevalent DUX4 inhibitors within HTS library Darko Bosnakovski1,2, Si Ho Choi2, Erik Toso2 , Michael A. Walters3, Jessica M. Strasser3 and Michael Kyba2 1University Goce Delcev - Stip, Faculty of Medical Sciences, Krste Misirkov b.b., 2000 Stip, R. Macedonia 2Lillehei Heart Institute, Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota, 55014 3Institute for Therapeutics Discovery and Development, College of Pharmacy, University of Minnesota, Minneapolis, Minnesota, 55014 DUX4 has emerged as the key molecular target in Facioscapulohumeral Muscular Dystrophy (FSHD). We have shown that DUX4 induces cell toxicity in skeletal muscle at high levels of expression while at low levels it interferes with differentiation and myogenic gene expression related to the FSHD molecular phenotype. A first step towards developing a targeted therapy for FSHD is to discover chemical compounds that inhibit the activity of the DUX4 protein. We have taken advantage of the conditional toxicity of DUX4-inducible myoblasts to develop a small molecule screening platform for identifying inhibitors of DUX4. The assay was based on rapid toxicity of high level DUX4 expression which leads to myoblast death within 24 hours. High throughput screening (HTS) of more than 200.000 small molecules as a part of UT Southwestern HTS compound library was done identified approximately 700 compounds with significant rescue effect. To identify direct inhibitors, we have conducted serial follow up assays, including secondary screens for interference with the conditional gene expression system, reversion of toxicity in other DUX4-expressing cell types (fibroblast, human ES cells) and protection against other cell death-inducing signals. We are currently working with a subset of 100 purchased compounds. These were selected based on pharmacological desirability analyses and the presence of multiple related compounds that form a chemical series within the hit set. While the majority of hits are singlets, our analysis identified 46 distinct chemical series. Rescreening these purchased compounds demonstrated Almost all of the purchased compounds demonstrated some degree of protection from DUX4-induced cell death indicating the success of the primary screen. Follow up screens indicated that the majority of these hits were also acting independently of the dox-inducible system. Interestingly, further analyses revealed that a very large subset of the selected compounds confer protection against an oxidative insult (tBHP). The data to date highlight the potential usefulness of antioxidant therapy in FSHD.

Item Type: Conference or Workshop Item (Lecture)
Subjects: Medical and Health Sciences > Basic medicine
Medical and Health Sciences > Clinical medicine
Divisions: Faculty of Medical Science
Depositing User: Darko Bosnakovski
Date Deposited: 29 Jan 2013 15:16
Last Modified: 29 Jan 2013 15:16

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