Monitor Ionizing Radiation-Induced Cellular Responses with Raman Spectroscopy,Non-Negative Matrix Factorization,and Non-Negative Least Squares

Radiation therapy (RT) is one of the most commonly prescribed cancer treatments. New tools that can accurately monitor and evaluate individual patient responses would be a major advantage and lend to the implementation of personalized treatment plans. In this study, Raman spectroscopy (RS) was applied to examine radiation-induced cellular responses in H460, MCF7, and LNCaP cancer cell lines across different dose levels and times post-irradiation. Previous Raman data analysis was conducted using principal component analysis (PCA), which showed the ability to extract biological information of glycogen. In the current studies, the use of non-negative matrix factorization (NMF) allowed for the discovery of multiplexed biological information, specifically uncovering glycogen-like and lipid-like component bases. The corresponding scores of glycogen and previously unidentified lipids revealed the content variations of these two chemicals in the cellular data. The NMF decomposed glycogen and lipid-like bases were able to separate the cancer cell lines into radiosensitive and radioresistant groups. A further lipid phenotype investigation was also attempted by applying non-negative least squares (NNLS) to the lipid-like bases decomposed individually from three cell lines. Qualitative differences found in lipid weights for each lipid-like basis suggest the lipid phenotype differences in the three tested cancer cell lines. Collectively, this study demonstrates that the application of NMF and NNLS on RS data analysis to monitor ionizing radiation-induced cellular responses can yield multiplexed biological information on bio-response to RT not revealed by conventional chemometric approaches.