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A Combination of screening and computational approaches for the identification of novel compounds that decrease mast cell degranulation

  • McShane, Marisa P. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
  • Friedrichson, Tim JADO Technologies GmbH, Dresden, Germany
  • Giner, Angelika Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
  • Meyenhofer, Felix Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany - University of Fribourg, Department of Medicine–Anatomy, Switzerland
  • Barsacchi, Rico Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
  • Bickle, Marc Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
  • Zerial, Marino Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
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    02.04.2015
Published in:
  • Journal of Biomolecular Screening. - 2015, p. 1087057115579613
English High-content screening of compound libraries poses various challenges in the early steps in drug discovery such as gaining insights into the mode of action of the selected compounds. Here, we addressed these challenges by integrating two biological screens through bioinformatics and computational analysis. We screened a small-molecule library enriched in amphiphilic compounds in a degranulation assay in rat basophilic leukemia 2H3 (RBL-2H3) cells. The same library was rescreened in a high-content image-based endocytosis assay in HeLa cells. This assay was previously applied to a genome-wide RNAi screen that produced quantitative multiparametric phenotypic profiles for genes that directly or indirectly affect endocytosis. By correlating the endocytic profiles of the compounds with the genome-wide siRNA profiles, we identified candidate pathways that may be inhibited by the compounds. Among these, we focused on the Akt pathway and validated its inhibition in HeLa and RBL-2H3 cells. We further showed that the compounds inhibited the translocation of the Akt-PH domain to the plasma membrane. The approach performed here can be used to integrate chemical and functional genomics screens for investigating the mechanism of action of compounds.
Faculty
Faculté des sciences et de médecine
Department
Département de Médecine
Language
  • English
Classification
Biology
License
License undefined
Identifiers
Persistent URL
https://folia.unifr.ch/unifr/documents/304121
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