Abstract
Non-invasively probing metabolites within single live cells is highly desired but challenging. Here we utilize Raman spectro-microscopy for spatial mapping of metabolites within single cells, with the specific goal of identifying druggable metabolic susceptibilities from a series of patient-derived melanoma cell lines. Each cell line represents a different characteristic level of cancer cell de-differentiation. First, with Raman spectroscopy, followed by stimulated Raman scattering (SRS) microscopy and transcriptomics analysis, we identify the fatty acid synthesis pathway as a druggable susceptibility for differentiated melanocytic cells. We then utilize hyperspectral-SRS imaging of intracellular lipid droplets to identify a previously unknown susceptibility of lipid mono-unsaturation within de-differentiated mesenchymal cells with innate resistance to BRAF inhibition. Drugging this target leads to cellular apoptosis accompanied by the formation of phase-separated intracellular membrane domains. The integration of subcellular Raman spectro-microscopy with lipidomics and transcriptomics suggests possible lipid regulatory mechanisms underlying this pharmacological treatment. Our method should provide a general approach in spatially-resolved single cell metabolomics studies.
Highlights
Non-invasively probing metabolites within single live cells is highly desired but challenging
The nature of cancer cell de-differentiation means that energetic requirements, cellular morphology, etc., are all altered, suggesting that cellular differentiation is accompanied by metabolic reprogramming[35]
Just like the well-reported phenotypic markers[28,36], metabolic genes showed a clear phenotype-dependent expression trend, with associated functions that span different metabolic processes (The representative metabolic genes are shown in the bottom of Fig. 1b, the complete heatmap and list of the top ranked metabolic genes are shown in Supplementary Fig. 1 and Supplementary Table 1)
Summary
Non-invasively probing metabolites within single live cells is highly desired but challenging. We utilize Raman spectro-microscopy for spatial mapping of metabolites within single cells, with the specific goal of identifying druggable metabolic susceptibilities from a series of patient-derived melanoma cell lines. We utilize hyperspectral-SRS imaging of intracellular lipid droplets to identify a previously unknown susceptibility of lipid mono-unsaturation within de-differentiated mesenchymal cells with innate resistance to BRAF inhibition. Drugging this target leads to cellular apoptosis accompanied by the formation of phase-separated intracellular membrane domains. We utilize Raman spectro-microscopy to characterize this series of related but distinct BRAF-mutant melanoma cancer cell phenotypes, each corresponding to a different level of cancer cell differentiation, from melanocytic (differentiated) to mesenchymal (de-differentiated)[25,26,27]. We mine the resulting spectroscopic information to identify phenotype specific, druggable metabolic susceptibilities
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