Imaging Mass Spectrometry Reveals Tumor Metabolic Heterogeneity.

Malignant tumors exhibit high degrees of genomic heterogeneity at the cellular level, leading to the view that subpopulations of tumor cells drive growth and treatment resistance. To examine the degree to which tumors also exhibit metabolic heterogeneity at the level of individual cells, we employed multi-isotope imaging mass spectrometry (MIMS) to quantify utilization of stable isotopes of glucose and glutamine along with a label for cell division. Mouse models of melanoma and malignant peripheral nerve sheath tumors (MPNSTs) exhibited striking heterogeneity of substrate utilization, evident in both proliferating and non-proliferating cells. We identified a correlation between metabolic heterogeneity, proliferation, and therapeutic resistance. Heterogeneity in metabolic substrate usage as revealed by incorporation of glucose and glutamine tracers is thus a marker for tumor proliferation. Collectively, our data demonstrate that MIMS provides a powerful tool with which to dissect metabolic functions of individual cells within the native tumor environment.

Imaging mass spectrometry reveals heterogeneity of proliferation and metabolism in atherosclerosis.

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.

Monitoring of in-season neuromuscular and perceptual fatigue in youth rugby players.

The purpose of this study was to examine both short- and long-term neuromuscular and perceptual fatigue in youth rugby players during a seven-week in-season mesocycle. Eleven male youth rugby players (age 16.9 ± 0.8 years) were assessed for countermovement jump (CMJ), reactive strength index (RSI) and leg stiffness to monitor neuromuscular performance, together with a well-being questionnaire to monitor perceptual fatigue. Players trained and competed throughout a seven-week block with test variables measured at baseline and 24 h pre- and post-matches played in weeks 1, 4 and 7. Players trained on average 9.7 ± 1.1 h per week and competed in 10.5 ± 1.9 games over the seven-week block. Pre- to post-match reductions were significant across all games for CMJ, RSI and well-being (all P < 0.05), ranging from likely to almost certain negative reductions. Well-being and RSI demonstrated non-significant (P < 0.05) unclear or inconsistent changes when comparing pre-match scores to baseline. Significant reductions to baseline were observed pre-match for CMJ (weeks 4 and 7) and stiffness (week 7), representing very likely to almost certain negative long-term decrements. CMJ, RSI and well-being were all sensitive to detecting post-match fatigue. Importantly, CMJ and stiffness were sensitive to detecting accumulated fatigue over a seven-week period, whereas RSI and well-being were not. Consequently, either a CMJ or leg stiffness should be monitored to detect long-term, accumulated fatigue in academy rugby players.