title: A dual-color fluorescence-based platform to identify selective inhibitors of Akt signaling.
abstract: BACKGROUND
  Inhibition of Akt signaling is considered one of the most promising therapeutic strategies for many cancers. However, rational target-orientated approaches to cell based drug screens for anti-cancer agents have historically been compromised by the notorious absence of suitable control cells.
  
  
  RESULTS
  In order to address this fundamental problem, we have developed BaFiso, a live-cell screening platform to identify specific inhibitors of this pathway. BaFiso relies on the co-culture of isogenic cell lines that have been engineered to sustain interleukin-3 independent survival of the parental Ba/F3 cells, and that are individually tagged with different fluorescent proteins. Whilst in the first of these two lines cell survival in the absence of IL-3 is dependent on the expression of activated Akt, the cells expressing constitutively-activated Stat5 signaling display IL-3 independent growth and survival in an Akt-independent manner. Small molecules can then be screened in these lines to identify inhibitors that rescue IL-3 dependence.
  
  
  CONCLUSIONS
  BaFiso measures differential cell survival using multiparametric live cell imaging and permits selective inhibitors of Akt signaling to be identified. BaFiso is a platform technology suitable for the identification of small molecule inhibitors of IL-3 mediated survival signaling.
authors: Experimental Therapeutics Program, Centro Nacional de Investigaciones Oncologicas, Madrid, Spain.
authors: Aranzazú
authors: A
authors: Rosado
authors: Fabian
authors: F
authors: Zanella
authors: Beatriz
authors: B
authors: Garcia
authors: Amancio
authors: A
authors: Carnero
authors: Wolfgang
authors: W
authors: Link
mesh_terms: D000818:Animals; D001483:Base Sequence; D018920:Coculture Techniques; D017931:DNA Primers; D005453:Fluorescence; D005456:Fluorescent Dyes; D007377:Interleukin-3 / Q000378:metabolism; D051379:Mice; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes; D007377:Interleukin-3; D047428:Protein Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt