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Analysis of Intracellular Fatty Acid Metabolism during Doxorubicin-Induced Senescence of MCF7 Cells Using Raman
Swarang Sachin Pundlik1, Ashwin Venkateshvaran1,2, Yavanica Suresh1
1Metabolic Regulation of Cell Fate (RCF), Institute for Stem Cell Science and Regenerative Medicine (BRIC-InStem), GKVK - Post, Bellary Road, Bengaluru 560065, Karnataka, India.
Abstract:
Cellular senescence, a stable growth-arrested state induced by stress or chemotherapeutic agents, is accompanied by metabolic remodeling that supports the senescence-associated secretory phenotype (SASP). Among these pathways, lipid and arachidonic acid (AA) metabolism play central roles in maintaining and propagating the senescent state. Here, we used hyperspectral confocal Raman microscopy to visualize biochemical remodeling in MCF7 human breast adenocarcinoma cells undergoing doxorubicin-induced senescence. Raman spectral analysis and principal component decomposition revealed time-dependent alterations in lipid-associated vibrational modes, particularly CH2 and CC stretching, consistent with enhanced lipid accumulation and remodeling between Days 10 and 15 after DNA damage induction. PCA of lipid-rich compartments isolated by using true component analysis also confirms progressive increases in triacylglycerol and unsaturated lipid signatures. Using deuterated arachidonic acid (AA-d 8) and COX2 inhibition, we further demonstrated real-time intracellular AA metabolism by tracking (CC)D stretching peaks (2220-2254 cm-1) in the Raman-silent window. The ratio of these deuterium bands to CH2 stretching provided a label-free quantitative metric for COX2-dependent AA turnover in senescent cells. Together, these findings establish hyperspectral Raman imaging as a powerful, nonperturbative tool to map lipid and oxylipin metabolism during cellular senescence, offering new avenues to identify metabolic vulnerabilities in senescent tumor cells.
Insights
Cellular senescence involves metabolic changes, particularly in lipid and arachidonic acid (AA) metabolism. Hyperspectral Raman imaging revealed lipid accumulation and AA turnover in senescent breast cancer cells, offering new insights into metabolic vulnerabilities.
Area of Science:
- Biochemistry
- Cell Biology
- Spectroscopy
Background:
- Cellular senescence is a state of stable growth arrest linked to metabolic changes.
- Lipid and arachidonic acid (AA) metabolism are crucial for maintaining the senescence-associated secretory phenotype (SASP).
Purpose of the Study:
- To visualize and quantify biochemical remodeling, specifically lipid and AA metabolism, in doxorubicin-induced senescent MCF7 breast cancer cells.
- To establish hyperspectral Raman imaging as a tool for studying senescence metabolism.
Main Methods:
- Hyperspectral confocal Raman microscopy was employed to analyze biochemical changes in senescent cells.
- Principal component analysis (PCA) and true component analysis were used for spectral data interpretation.
- Deuterated arachidonic acid (AA-d8) and COX2 inhibition were utilized to track AA metabolism in real-time.
Main Results:
- Raman spectral analysis showed time-dependent alterations in lipid metabolism, with increased lipid accumulation and remodeling between Days 10-15.
- PCA confirmed progressive increases in triacylglycerol and unsaturated lipid signatures in senescent cells.
- A label-free quantitative metric was developed to measure COX2-dependent AA turnover using Raman spectroscopy.
Conclusions:
- Hyperspectral Raman imaging is a powerful, nonperturbative method for mapping lipid and oxylipin metabolism during cellular senescence.
- This technique can reveal metabolic vulnerabilities in senescent tumor cells, potentially leading to new therapeutic strategies.
