An integrated one-step system to extract, analyze and annotate all relevant information from image-based cell screening of chemical libraries.
Here we report the development and validation of a complete solution to manage and analyze the data produced by image-based phenotypic screening campaigns of small-molecule libraries. In one step initial crude images are analyzed for multiple cytological features, statistical analysis is performed and molecules that produce the desired phenotypic profile are identified. A naïve Bayes classifier, integrating chemical and phenotypic spaces, is built and utilized during the process to assess those images initially classified as "fuzzy"-an automated iterative feedback tuning. Simultaneously, all this information is directly annotated in a relational database containing the chemical data. This novel fully automated method was validated by conducting a re-analysis of results from a high-content screening campaign involving 33 992 molecules used to identify inhibitors of the PI3K/Akt signaling pathway. Ninety-two percent of confirmed hits identified by the conventional multistep analysis method were identified using this integrated one-step system as well as 40 new hits, 14.9% of the total, originally false negatives. Ninety-six percent of true negatives were properly recognized too. A web-based access to the database, with customizable data retrieval and visualization tools, facilitates the posterior analysis of annotated cytological features which allows identification of additional phenotypic profiles; thus, further analysis of original crude images is not required.
Experimental Therapeutics Programme, Spanish National Cancer Research Centre (CNIO), Melchor Fernandez Almagro, 3, 28029 Madrid, Spain.
Obdulia
O
Rabal
Wolfgang
W
Link
Beatriz G
BG
Serelde
James R
JR
Bischoff
Julen
J
Oyarzabal
D002460:Cell Line; D003628:Database Management Systems; D004353:Drug Evaluation, Preclinical / Q000706:statistics & numerical data*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000706:statistics & numerical data*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors; D015398:Signal Transduction / Q000187:drug effects; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000494:pharmacology*; D049490:Systems Biology
D004791:Enzyme Inhibitors; D000081082:Phosphoinositide-3 Kinase Inhibitors; D054852:Small Molecule Libraries; D051057:Proto-Oncogene Proteins c-akt