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Imaging mass spectrometry reveals heterogeneity of proliferation and metabolism in atherosclerosis
Christelle Guillermier, … , Matthew L. Steinhauser, Jonathan D. Brown
Christelle Guillermier, … , Matthew L. Steinhauser, Jonathan D. Brown
Published June 6, 2019
Citation Information: JCI Insight. 2019;4(11):e128528. https://doi.org/10.1172/jci.insight.128528.
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Technical Advance Vascular biology

Imaging mass spectrometry reveals heterogeneity of proliferation and metabolism in atherosclerosis

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Abstract

Atherosclerotic plaques feature local proliferation of leukocytes and vascular smooth muscle cells (VSMCs) and changes in cellular metabolism. Yet the relationship between glucose utilization and proliferation has been technically impossible to study directly in cells of atherosclerotic plaques in vivo. We used multi-isotope imaging mass spectrometry (MIMS), a quantitative imaging platform, to measure coincident cell division and glucose utilization at suborganelle resolution in atherosclerotic plaques. In established plaques, 65% of intimal foam cells and only 4% of medial VSMCs were labeled with 15N-thymidine after 1 week of isotope treatment. Dividing cells demonstrated heightened glucose labeling. MIMS detected 2H-glucose label in multiple subcellular compartments within foam cells, including lipid droplets, the cytosol, and chromatin. Unexpectedly, we identified an intensely focal region of 2H-label in VSMCs underlying plaques. This signal diminished in regions of aorta without atherosclerosis. In advanced plaques, 15N-thymidine and 2H-glucose labeling in foam cells and VSMCs significantly decreased. These data demonstrate marked heterogeneity in VSMC glucose metabolism that was dependent on both proliferative status and proximity of VSMCs to plaques. Furthermore, these results reveal how quantitative mass spectrometry coupled with isotope imaging can complement other methods used to study cell biology directly in the growing atherosclerotic plaque in vivo.

Authors

Christelle Guillermier, Sean P. Doherty, Adam G. Whitney, Vladimir R. Babaev, MacRae F. Linton, Matthew L. Steinhauser, Jonathan D. Brown

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Figure 2

Subcellular glucose utilization is enriched in proliferating foam cells.

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Subcellular glucose utilization is enriched in proliferating foam cells....
(A) Intimal foam cells. These images reveal higher-intensity 2H-glucose labeling in divided 15N+ cells (white arrows, cell border). Scale bar: 5 μm. (B) 2H-glucose labeling distributions in ECs and foam cells from 3 mice. Each dot denotes an individual cell. Bars denote median ± interquartile range. (C) 2H-glucose labeling distributions of nucleated foam cells as a function of whether they divided (15N+) during the labeling period. Data are expressed relative to the undivided (15N–) fraction. Nucleated intimal cells from n = 3 mice pooled. Each dot denotes an individual cell. Bars denote mean ± SD. Significance was assessed by unpaired t test. (D) Mapping of 2H-glucose labeling in subcellular domains. Regions of interest (ROI) corresponding to individual lipid droplets (which appear black in 12C14N–), cytosolic regions between lipid droplets, and chromatin regions (31P– bright) were selected using the 12C14N– and 31P– images. 2H-glucose labeling data (2H/1H ratio) was extracted for each ROI and is expressed in the graph (right) as relative to background. Dashed lines correspond to background (1) plus 2 standard deviations (2σ). Black line denotes median for each distribution. Significance was assessed by ANOVA; P < 0.0001.
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