A Unique High-Throughput Assay to Identify Novel Small Molecule Inhibitors of Chemotaxis and Migration.
A Unique High-Throughput Assay to Identify Novel Small Molecule Inhibitors of Chemotaxis and Migration.
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DOI：
10.1002/cpcb.17
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发表时间：
2017-03-03
影响因子：
--
通讯作者：
Kimmel AR
中科院分区：
文献类型：
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作者：
Liao XH;Kimmel AR
趋化性和细胞迁移在正常的生理过程如胚胎发生、炎症和伤口愈合中以及在病理过程包括慢性炎性疾病和癌症转移中起关键作用。新型趋化性/迁移抑制剂对于开发有效的治疗剂和探测分子机制是期望的。我们描述了一种基于荧光的表型测定在1536孔板格式的高通量筛选的新型抑制剂的趋化性/迁移复杂的图书馆内的多1000化合物。尽管该测定利用了网骨藻独特的细胞反应特性，但所鉴定的化合物能够抑制哺乳动物细胞的趋化性。此外，一个平行的细胞毒性计数屏幕与ATP含量测定的描述，消除细胞毒性化合物从屏幕上。这种新的化合物筛选方法能够快速鉴定抑制人类和其他细胞趋化性的新型先导化合物，用于药物开发和研究工具。
Chemotaxis and cell migration play pivotal roles in normal physiological processes such as embryogenesis, inflammation, and wound healing, as well as, in pathological processes including chronic inflammatory disease and cancer metastasis. Novel chemotaxis/migration inhibitors are desirable for developing effective therapeutics and probing molecular mechanisms. We describe a fluorescence-based phenotypic assay in a 1536-well plate format for high-throughput screening of novel inhibitors of chemotaxis/migration within complex libraries of multi-1000 compounds. Although the assay utilizes the unique cellular response properties of Dictyostelium, the compounds identified are able to inhibit chemotaxis of mammalian cells. In addition, a parallel cell cytotoxicity counter-screen with an ATP content assay is described that eliminates cytotoxic compounds from the screen. This novel compound screening approach enables rapid identification of novel lead compounds that inhibit chemotaxis in human and other cells for drug development and research tools.