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Rapid recognition of drug-resistance/sensitivity in leukemic cells by Fourier transform InfraRed microspectroscopy and unsupervised Hierarchical Cluster Analysis.

DOI: 10.1039/c2an36393c DOI Help

Authors: Giuseppe Bellisola (Azienda Ospedaliera Universitaria Integrata Verona (AOUI)) , Gianfelice Cinque (Diamond Light Source) , Marzia Vezzalini (University of Verona) , Elisabetta Moratti (University of Verona) , Giovannino Silvestri (University of Verona) , Sara Redaelli (University of Milano-Bicocca) , Carlo Gambacorti Passerini (University of Milano-Bicocca) , Katia Wehbe (Diamond Light Source) , Claudio Sorio (University of Verona)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Analyst , VOL 138 (14)

State: Published (Approved)
Published: January 2013
Diamond Proposal Number(s): 6675 , 7143

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 p210BCR-ABL drug-sensitive wild-type BCR-ABL or the V299L or T315I p210BCR-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 ?m2 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.

Subject Areas: Biology and Bio-materials

Instruments: B22-Multimode InfraRed imaging And Microspectroscopy