title: High-throughput high-content imaging assays for identification and characterization of selective AXL pathway inhibitors.
abstract: Receptor tyrosine kinases (RTKs) regulate a wide range of important biological activities, including cell proliferation, differentiation, migration, and apoptosis. Abnormalities in RTKs are involved in numerous diseases, including cancer and other proliferative disorders. AXL belongs to the TAM (Tyso3, AXL, and Mer) family of RTKs. The AXL signaling pathway represents an attractive target for the treatment of diseases, such as cancer. Using phospho-AKT as readout, a high-throughput 384-well cell-based assay was established in the NCI-H1299 human non-small cell lung carcinoma cell line to evaluate compound potency in inhibiting AXL pathway activation. In addition, a counter screen assay was established in the same cellular background to differentiate AXL kinase inhibitors from AXL receptor antagonists, which block the interaction of AXL and its natural ligand GAS6. These cell-based functional assays are useful tools in the identification and optimization of small molecules and biological reagents for potential therapeutics for the treatment of GAS6/AXL-related diseases.
authors.affiliation: 1 Department of Leads Discovery and Optimization, Bristol-Myers Squibb , Princeton, New Jersey.
authors.forename: Huaping
authors.initials: H
authors.lastname: Tang
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.forename: Ding Ren
authors.initials: DR
authors.lastname: Shen
authors.forename: Deepa
authors.initials: D
authors.lastname: Calambur
authors.forename: Mark
authors.initials: M
authors.lastname: Witmer
authors.forename: Sophie
authors.initials: S
authors.lastname: Wu
authors.forename: Brian
authors.initials: B
authors.lastname: Carpenter
authors.forename: Yaqun
authors.initials: Y
authors.lastname: Zhang
authors.forename: Mian
authors.initials: M
authors.lastname: Gao
authors.forename: Keith
authors.initials: K
authors.lastname: Constantine
authors.forename: Litao
authors.initials: L
authors.lastname: Zhang
authors.forename: Mary Ellen
authors.initials: ME
authors.lastname: Cvijic
mesh_terms: D001681:Biological Assay / Q000379:methods; D002289:Carcinoma, Non-Small-Cell Lung / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004353:Drug Evaluation, Preclinical / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008175:Lung Neoplasms / Q000378:metabolism* / Q000473:pathology; D008856:Microscopy, Fluorescence / Q000379:methods; D057054:Molecular Imaging / Q000379:methods; D047428:Protein Kinase Inhibitors / Q000008:administration & dosage*; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D000094963:Axl Receptor Tyrosine Kinase
chemical_list: D047428:Protein Kinase Inhibitors; D011518:Proto-Oncogene Proteins; D020794:Receptor Protein-Tyrosine Kinases; D000094963:Axl Receptor Tyrosine Kinase; C000722289:AXL protein, human