Marković Ivana, Lukić Iva, Lukić Maja, Pavičić Ema, Mrđenović Stefan, Kotris Ana, Dmitrović Branko, Debeljak Željko
Clinical Institute of Laboratory Diagnostics, University Hospital Centre Osijek, J. Huttlera 4, Osijek 31 000, Croatia; Faculty of Medicine Osijek, JJ Strossmayer University of Osijek, J. Huttlera 4, Osijek 31 000, Croatia.
Faculty of Food Technology Osijek, JJ Strossmayer University of Osijek, F. Kuhača 18, Osijek 31 000, Croatia.
J Pharm Biomed Anal. 2025 Mar 15;255:116664. doi: 10.1016/j.jpba.2025.116664. Epub 2025 Jan 5.
Ibrutinib and acalabrutinib, Bruton's tyrosine kinase inhibitors (BTKi) used for chronic lymphocytic leukemia (CLL) treatment, aim the same target but their off-target effects are different. The aim of this study was to use single-cell MALDI TOF mass spectrometry imaging to compare the CD19+ lymphocytes' mass spectra in untreated and ibrutinib- or acalabrutinib-treated subjects in order to better understand the therapeutic effect of BTKi. 180 cells from 9 male subjects divided in 3 groups (untreated, ibrutinib-treated and acalabrutinib-treated) were analyzed using MALDI-TOF mass spectrometry analyzer. Mass spectra were acquired in the 300-600 Da mass range. Partial least squares discriminant analysis (PLS-DA) was used for evaluation of mass spectra, while Volcano plots and Venn diagram were used for a review of significantly altered m/z values. Cross-validated PLS-DA classification accuracy of cells was 85 %. Ibrutinib-treated cells overlap with the untreated cell population, while acalabrutinib-treated cells form a separate class. 13 m/z signals were specific for each BTKi group. In conclusion, single-cell MALDI-TOF mass spectrometry imaging detects differences in the mass spectra of CD19+ lymphocytes treated with two different BTKi. Drug-specific m/z signal clusters can be identified as a chemical fingerprint of the BTKi effect and represent candidates for the therapeutic response biomarkers.
