title: Rapid recognition of drug-resistance/sensitivity in leukemic cells by Fourier transform infrared microspectroscopy and unsupervised hierarchical cluster analysis.
abstract: We tested the ability of Fourier Transform (FT) InfraRed (IR) microspectroscopy (microFTIR) in combination with unsupervised Hierarchical Cluster Analysis (HCA) in identifying drug-resistance/sensitivity in leukemic cells exposed to tyrosine kinase inhibitors (TKIs). Experiments were carried out in a well-established mouse model of human Chronic Myelogenous Leukemia (CML). Mouse-derived pro-B Ba/F3 cells transfected with and stably expressing the human p210(BCR-ABL) drug-sensitive wild-type BCR-ABL or the V299L or T315I p210(BCR-ABL) drug-resistant BCR-ABL mutants were exposed to imatinib-mesylate (IMA) or dasatinib (DAS). MicroFTIR was carried out at the Diamond IR beamline MIRIAM where the mid-IR absorbance spectra of individual Ba/F3 cells were acquired using the high brilliance IR synchrotron radiation (SR) via aperture of 15 × 15 μm(2) in sizes. A conventional IR source (globar) was used to compare average spectra over 15 cells or more. IR signatures of drug actions were identified by supervised analyses in the spectra of TKI-sensitive cells. Unsupervised HCA applied to selected intervals of wavenumber allowed us to classify the IR patterns of viable (drug-resistant) and apoptotic (drug-sensitive) cells with an accuracy of >95%. The results from microFTIR + HCA analysis were cross-validated with those obtained via immunochemical methods, i.e. immunoblotting and flow cytometry (FC) that resulted directly and significantly correlated. We conclude that this combined microFTIR + HCA method potentially represents a rapid, convenient and robust screening approach to study the impact of drugs in leukemic cells as well as in peripheral blasts from patients in clinical trials with new anti-leukemic drugs.
authors: Azienda Ospedaliera Universitaria Integrata Verona, Department of Pathology and Diagnostics - Unit of Immunology, Policlinico G. Rossi, P.le L.A. Scuro, 10, I-37134, Verona, Italy. giuseppe.bellisola@univr.it
authors: Giuseppe
authors: G
authors: Bellisola
authors: Gianfelice
authors: G
authors: Cinque
authors: Marzia
authors: M
authors: Vezzalini
authors: Elisabetta
authors: E
authors: Moratti
authors: Giovannino
authors: G
authors: Silvestri
authors: Sara
authors: S
authors: Redaelli
authors: Carlo
authors: C
authors: Gambacorti Passerini
authors: Katia
authors: K
authors: Wehbe
authors: Claudio
authors: C
authors: Sorio
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D016000:Cluster Analysis; D000069439:Dasatinib; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D016044:Fusion Proteins, bcr-abl / Q000037:antagonists & inhibitors / Q000235:genetics*; D006801:Humans; D000068877:Imatinib Mesylate; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000175:diagnosis* / Q000188:drug therapy; D051379:Mice; D009154:Mutation / Q000235:genetics*; D010879:Piperazines / Q000494:pharmacology; D054448:Precursor Cells, B-Lymphoid / Q000187:drug effects / Q000473:pathology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013844:Thiazoles / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D001549:Benzamides; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D013844:Thiazoles; D000068877:Imatinib Mesylate; D016044:Fusion Proteins, bcr-abl; D000069439:Dasatinib