Tracking InfraRed signatures of drugs in cancer cells by Fourier Transform microspectroscopy

• Published in: Analyst (London) Vol. 135; no. 12; pp. 3077 - 3086
• Main Authors: BELLISOLA, Giuseppe, DELLA PERUTA, Marco, VEZZALINI, Marzia, MORATTI, Elisabetta, VACCARI, Lisa, BIRARDA, Giovanni, PICCININI, Massimo, CINQUE, Gianfelice, SORIO, Claudio
• Format: Conference Proceeding Journal Article
• Language: English
• Published: Cambridge Royal Society of Chemistry 01.12.2010
• Subjects: Ageing, cell death; Analytical chemistry; Antineoplastic Agents - chemistry; Antineoplastic Agents - therapeutic use; Apoptosis; Biological and medical sciences; Cancer; Cell physiology; Cell transformation and carcinogenesis. Action of oncogenes and antioncogenes; Chemistry; Cluster Analysis; Drug Resistance, Neoplasm; Drugs; Exact sciences and technology; Fourier transforms; Fundamental and applied biological sciences. Psychology; Humans; Infrared signatures; Infrared spectra; K562 Cells; Kinases; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - drug therapy; Mathematical analysis; Mathematical models; Microscopy - instrumentation; Microscopy - methods; Molecular and cellular biology; Proteins - chemistry; Reproducibility of Results; Signatures; Spectrometric and optical methods; Spectroscopy, Fourier Transform Infrared - instrumentation; Spectroscopy, Fourier Transform Infrared - methods; Cluster analysis; Monolayer; Fourier transform spectroscopy; Chemical analysis; Targeting; Leukemia; Identification; Molecular targeting; Carcinogenesis; Spectral data; Sample preparation; Cadmium tellurides; Mercury tellurides; Anomaly; Diagnosis; Synchrotron radiation; Drug; Tyrosine; Sample; Pattern recognition; Protein; Infrared spectrometry; Screening; Absorption spectrometry; Detector; Cell line; Analytical method; Frequency; Parameter; Technique; Spatial resolution; Apoptosis
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/c0an00509f

Aimed at developing accurate, reliable and cost-saving analytical techniques for drugs screening we evaluated the potential of Fourier Transform (FT) InfraRed (IR) microspectroscopy (microFTIR) as a quantitative pre-diagnostic approach for the rapid identification of IR signatures of drugs targeting specific molecular pathways causing Chronic Myeloid Leukemia (CML). To obtain reproducible FTIR absorbance spectra at the necessary spatial resolution we optimized sample preparation and acquisition parameters on a single channel Mercury-Cadmium-Telluride (MCT) detector in the spectral interval of frequencies from 4000 to 800 cm(-1). Single K562 cells were illuminated by Synchrotron Radiation (SR) and a number of ~15 K562 cells spread in monolayer were illuminated by a conventional IR source (Globar), respectively. Combining IR spectral data with the results of complementary biochemical investigations carried out in samples by different analytical methods we identified and cross-validated IR signatures of drugs targeting the oncogenic protein BCR/ABL and its associated abnormal tyrosine kinase activity in K562 cell line. Unsupervised pattern recognition performed by Hierarchical Cluster Analysis (HCA) clustered the spectra of single K562 cells in two distinct groups roughly corresponding to living and to apoptotic cells, respectively. The corresponding IR spectral profiles were assumed to represent drug-resistant and drug-sensitive cells. Significant variations with increasing percentages of apoptotic cells were observed after the treatment of K562 cells with drugs that directly or indirectly target BCR/ABL. In conclusion, we suggest that microFTIR associated with multivariate data analysis may be useful to assess drug compounds in ex vivo cancer cell models and possibly peripheral blast cells from CML patients.