title: Comparison of Calcium Dynamics and Specific Features for G Protein-Coupled Receptor-Targeting Drugs Using Live Cell Imaging and Automated Analysis.
abstract: G protein-coupled receptors (GPCRs) are targets for designing a large fraction of the drugs in the pharmaceutical industry. For GPCR-targeting drug screening using cell-based assays, measurement of cytosolic calcium has been widely used to obtain dose-response profiles. However, it remains challenging to obtain drug-specific features due to cell-to-cell heterogeneity in drug-cell responses obtained from live cell imaging. Here, we present a framework combining live cell imaging of a cell population and a feature extraction method for classification of responses of drugs targeting GPCRs CXCR4 and α2AR. We measured the calcium dynamics using confocal microscopy and compared the responses for SDF-1α and norepinephrine. The results clearly show that the clustering patterns of responses for the two GPCRs are significantly different. Additionally, we show that different drugs targeting the same GPCR induce different calcium response signatures. We also implemented principal component analysis and k means for feature extraction and used nondominated (ND) sorting for ranking a group of drugs at various doses. The presented approach can be used to model a cell population as a mixture of subpopulations. It also offers specific advantages, such as higher spatial resolution, classification of responses, and ranking of drugs, potentially providing a platform for high-content drug screening.
authors.affiliation: 1 Department of Chemical Engineering, Indian Institute of Technology-Hyderabad, Hyderabad, Telangana, India.
authors.forename: Rishikesh Kumar
authors.initials: RK
authors.lastname: Gupta
authors.affiliation: 1 Department of Chemical Engineering, Indian Institute of Technology-Hyderabad, Hyderabad, Telangana, India.
authors.forename: Sarpras
authors.initials: S
authors.lastname: Swain
authors.affiliation: 2 Department of Chemistry and Biochemistry, The University of Toledo, Toledo, OH, USA.
authors.forename: Dinesh
authors.initials: D
authors.lastname: Kankanamge
authors.affiliation: 1 Department of Chemical Engineering, Indian Institute of Technology-Hyderabad, Hyderabad, Telangana, India.
authors.forename: Pantula Devi
authors.initials: PD
authors.lastname: Priyanka
authors.affiliation: 1 Department of Chemical Engineering, Indian Institute of Technology-Hyderabad, Hyderabad, Telangana, India.
authors.forename: Ranjana
authors.initials: R
authors.lastname: Singh
authors.affiliation: 1 Department of Chemical Engineering, Indian Institute of Technology-Hyderabad, Hyderabad, Telangana, India.
authors.forename: Kishalay
authors.initials: K
authors.lastname: Mitra
authors.affiliation: 2 Department of Chemistry and Biochemistry, The University of Toledo, Toledo, OH, USA.
authors.forename: Ajith
authors.initials: A
authors.lastname: Karunarathne
authors.affiliation: 1 Department of Chemical Engineering, Indian Institute of Technology-Hyderabad, Hyderabad, Telangana, India.
authors.forename: Lopamudra
authors.initials: L
authors.lastname: Giri
mesh_terms: D002118:Calcium / Q000378:metabolism*; D045744:Cell Line, Tumor; D003600:Cytosol / Q000378:metabolism; D016503:Drug Delivery Systems / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D025341:Principal Component Analysis / Q000379:methods; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*
chemical_list: D043562:Receptors, G-Protein-Coupled; D002118:Calcium
keywords: G protein–coupled receptor (GPCR); cell-to-cell heterogeneity; confocal microscopy; high-content screening; live cell calcium imaging