Analysis of lipids by Raman spectroscopy and mass spectrometry provides a detection tool and mechanistic insight into imatinib resistance in CML-BC
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Updated: May 28, 2025
An Integrated Raman Spectroscopy and Mass Spectrometry Platform to Study Single-Cell Drug Uptake, Metabolism, and Effects
Published on: January 9, 2020
Manish Bhat1, Panchali Saha1, Mythreyi Narasimhan2
1Advanced Centre for Treatment, Research and Education in Cancer, Tata Memorial Centre, Navi Mumbai 410210, India; Homi Bhabha National Institute, BARC Training School Complex, Mumbai 400094, India.
Abstract:
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.
Insights
Detecting tyrosine kinase inhibitor (TKI) resistance in chronic myeloid leukemia (CML) blast crisis (BC) is crucial. This study used lipid analysis via Raman spectroscopy and mass spectrometry to rapidly detect TKI resistance and uncover underlying mechanisms.
Area of Science:
- Biochemistry
- Oncology
- Analytical Chemistry
Background:
- Tyrosine kinase inhibitor (TKI) resistance poses a significant challenge in treating chronic myeloid leukemia (CML), particularly in the blast crisis (BC) phase.
- Existing resistance detection methods are limited in the BC phase due to incompletely understood resistance mechanisms.
- Novel approaches are needed to detect TKI resistance irrespective of the mechanism and to elucidate resistance pathways for targeted therapy.
Purpose of the Study:
- To develop and validate methods for rapid detection of TKI resistance in CML-BC.
- To identify and understand the lipid alterations associated with TKI resistance in CML-BC.
- To explore the mechanistic link between lipid changes and TKI resistance.
Main Methods:
- Lipid analysis of TKI-sensitive and TKI-resistant CML-BC cells using Raman spectroscopy and mass spectrometry.
- Application of Principal Component Analysis (PCA) and Principal Component based Linear Discriminant Analysis (PC-LDA) for spectral data analysis.
- Quantitative differential lipid profiling and mechanistic studies involving phospholipase A2 (PLA2) and p38 mitogen activated protein kinase (p38MAPK).
Main Results:
- Raman spectroscopy successfully differentiated TKI-resistant from TKI-sensitive CML-BC cells based on lipid spectral signatures.
- Mass spectrometry provided quantitative differential lipid profiles in resistant cells, revealing alterations in diverse lipid classes.
- These lipid alterations were linked to the modulation of PLA2 levels mediated by p38MAPK, establishing a causal association with TKI resistance.
Conclusions:
- Lipid analysis using complementary techniques like Raman spectroscopy and mass spectrometry offers a viable strategy for rapid TKI resistance detection in CML-BC.
- The study elucidated novel lipid alterations and their mechanistic connection to TKI resistance, involving the PLA2/p38MAPK pathway.
- These findings contribute to both the detection and mechanistic understanding of imatinib resistance in CML-BC, paving the way for improved therapeutic strategies.
