Analysis of lipids by Raman spectroscopy and mass spectrometry provides a detection tool and mechanistic insight into imatinib resistance in CML-BC

• Published in: Biochimica et biophysica acta. General subjects Vol. 1869; no. 4; p. 130771
• Main Authors: Bhat, Manish, Saha, Panchali, Narasimhan, Mythreyi, Shelar, Ashutosh, Hole, Arti, Murali Krishna, C., Govekar, Rukmini
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2025
• Subjects: Blast Crisis - drug therapy; Blast Crisis - metabolism; Blast Crisis - pathology; Cell Line, Tumor; Chronic myeloid leukemia; discriminant analysis; Drug Resistance, Neoplasm; drugs; Humans; Imatinib Mesylate - pharmacology; Imatinib Mesylate - therapeutic use; Imatinib resistance; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - drug therapy; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - metabolism; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - pathology; Lipids; Lipids - analysis; Mass spectrometry; Mass Spectrometry - methods; mitogen-activated protein kinase; myeloid leukemia; phospholipases; Principal Component Analysis; Protein Kinase Inhibitors - pharmacology; Protein Kinase Inhibitors - therapeutic use; Raman spectroscopy; Spectrum Analysis, Raman - methods; tyrosine; Lipids; Chronic myeloid leukemia; Imatinib resistance; Raman spectroscopy; Mass spectrometry
• ISSN: 0304-4165; 1872-8006; 1872-8006
• DOI: 10.1016/j.bbagen.2025.130771

Resistance to tyrosine kinase inhibitors (TKIs) is a major challenge in the treatment of chronic myeloid leukemia (CML). Established tests based on the known mechanisms of resistance in the initial chronic phase (CP) confirm resistance, reveal the underlying reason and thereby direct treatment modifications. In the terminal phase of blast crisis (BC), however, additional partially identified mechanisms of resistance exist which necessitates developing modalities to detect resistance regardless of the underlying mechanism and concurrent exploration of the resistance mechanism to assist in identification of appropriate drug targets. In this study both the clinical objectives were achieved by analysing lipids in BC cells, sensitive and resistant to TKIs, using the complementary strengths of distinct analytical technologies. Raman spectroscopy, through the spectral signatures with lipids as a significant differentiating component could segregate resistant from sensitive cells in the Principal Component Analysis (PCA) and Principal Component based Linear Discriminant Analysis (PC-LDA). This provided a tool to rapidly detect resistance in CML-BC despite unclear mechanism of TKI resistance. The depth of coverage achievable by mass spectrometry allowed the generation of quantitative differential profile of individual lipids in resistant cells. The alterations were in diverse classes of lipids which are involved in cell signalling and inhibition studies could link these alterations to modulation of phospholipase A2 (PLA2) levels mediated by p38 mitogen activated protein kinase (p38MAPK), which is causally associated with TKI resistance in CML-BC. Together, lipid analysis using the two platforms, contributed to the detection and mechanistic understanding of imatinib resistance in CML-BC. [Display omitted] •Mass spectrometry identifies altered lipid profile in imatinib-resistant CML cells.•Changes in signalling lipids linked to known mechanism of TKI resistance in CML-BC.•Lipid features of Raman spectra predominate in discriminating resistant cells.•Raman spectroscopy has potential to detect resistance regardless of the mechanism.