Kinetic assay shows that increasing red cell volume could be a treatment for sickle cell disease
Kinetic assay shows that increasing red cell volume could be a treatment for sickle cell disease
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DOI：
10.1073/pnas.1619054114
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发表时间：
2017-01-31
影响因子：
11.1
通讯作者：
Eaton, William A.
中科院分区：
文献类型：
--
作者：
Li, Quan;Henry, Eric R.;Eaton, William A.
尽管60多年来人们已经知道镰状细胞病的原因是血红蛋白突变体的聚合，但羟基脲是美国食品和药物管理局批准用于治疗的唯一药物。然而，这种药物只是部分成功，并且由于缺乏灵敏和定量的细胞测定，抑制纤维形成的其他药物的发现受到阻碍。在这里，我们描述了这样一种方法，在96孔板格式，是基于激光诱导聚合的镰状性状细胞和强大的，自动化的图像分析，以检测精确的时间，在纤维扭曲（“镰刀”）的细胞。通过这种动力学方法，我们发现，细胞体积的小幅增加可以降低血红蛋白浓度，从而导致纤维形成前延迟时间的治疗性增加。我们还表明，在临床试验中通过增加氧亲和力抑制聚合的两种药物（AES103和GBT 440）中，其中一种（GBT 440）还通过两种额外的机制抑制缺氧条件下的镰状化。
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 kineticmethod, 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.