Author:
Li Quan,Henry Eric R.,Hofrichter James,Smith Jeffrey F.,Cellmer Troy,Dunkelberger Emily B.,Metaferia Belhu B.,Jones-Straehle Stacy,Boutom Sarah,Christoph Garrott W.,Wakefield Terri H.,Link Mary E.,Staton Dwayne,Vass Erica R.,Miller Jeffery L.,Hsieh Matthew M.,Tisdale John F.,Eaton William A.
Abstract
Although it has been known for more than 60 years that the cause of sickle cell disease is polymerization of a hemoglobin mutant, hydroxyurea is the only drug approved for treatment by the US Food and Drug Administration. This drug, however, is only partially successful, and the discovery of additional drugs that inhibit fiber formation has been hampered by the lack of a sensitive and quantitative cellular assay. Here, we describe such a method in a 96-well plate format that is based on laser-induced polymerization in sickle trait cells and robust, automated image analysis to detect the precise time at which fibers distort (“sickle”) the cells. With this kinetic method, we show that small increases in cell volume to reduce the hemoglobin concentration can result in therapeutic increases in the delay time prior to fiber formation. We also show that, of the two drugs (AES103 and GBT440) in clinical trials that inhibit polymerization by increasing oxygen affinity, one of them (GBT440) also inhibits sickling in the absence of oxygen by two additional mechanisms.
Publisher
Proceedings of the National Academy of Sciences
Cited by
42 articles.
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