In vivo analysis of compound activity and mechanism of action using epistasis in Drosophila
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ORIGINAL ARTICLE
In vivo analysis of compound activity and mechanism of action using epistasis in Drosophila Erdem Bangi & Dan Garza & Marc Hild
Received: 28 May 2010 / Accepted: 30 November 2010 / Published online: 22 December 2010 # Springer-Verlag 2010
Abstract The recent establishment of high-throughput methods for culturing Drosophila provided a unique ability to screen compound libraries against complex disease phenotypes in the context of whole animals. However, as compound studies in Drosophila have been limited so far, the degree of conservation of compound activity between Drosophila and vertebrates or the effectiveness of feeding as a compound delivery system is not well known. Our comprehensive in vivo analysis of 27 small molecules targeting seven signaling pathways in Drosophila revealed a high degree of conservation of compound activity between Drosophila and vertebrates. We also investigated the mechanism of action of AY9944, one of the Hh pathway antagonists that we identified in our compound feeding experiments. Our epistasis analysis of AY9944 provided novel insights into AY9944’s mechanism of action and revealed a novel role for cholesterol transport in Hh signal transduction. Electronic supplementary material The online version of this article (doi:10.1007/s12154-010-0051-5) contains supplementary material, which is available to authorized users. M. Hild Developmental and Molecular Pathways, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA Present Address: D. Garza Proteostasis Therapeutics, Cambridge, MA 02139, USA Present Address: E. Bangi (*) Developmental and Regenerative Biology, Mount Sinai Medical Center, New York, NY 10029, USA e-mail: [email protected]
Keywords Chemical genetics . Compound feeding . Drosophila . Drug discovery . Signal transduction
Introduction In recent years, there has been growing interest in utilizing the genetic power of invertebrate model systems such as Drosophila in drug discovery as they provide highthroughput screening-compatible platforms that allow compound testing in a physiologically relevant environment [39]. Drosophila is a particularly attractive screening tool in this regard as many complex disease states associated with physiological defects in humans have been successfully modeled in flies, and these models mimic the pathology and specific symptoms of the diseases that are not easy to recapitulate in cell-based or in vitro assays [3, 5, 23, 25, 29]. Moreover, the reduced redundancy of the Drosophila genome could allow the identification of novel druggable targets that may be missed due to partially redundant activities of homologous proteins usually present in mammalian cells. Lastly, the availability of a large number of genetic tools allows rapid and detailed studies of compound activity in various mutant backgrounds and can provide novel insights into the compounds’ mechanism of action. Only a small number of compound studies in Drosophila have been reported so far, mostly focusing on analyses of individual compounds [3
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