Mammalian tissues are fuelled by circulating nutrients, including glucose, amino acids, and various intermediary metabolites. Under aerobic conditions, glucose is generally assumed to be burned fully by tissues via the tricarboxylic acid cycle (TCA cycle) to carbon dioxide. Alternatively, glucose can be catabolized anaerobically via glycolysis to lactate, which is itself also a potential nutrient for tissues1 and tumours2,3,4,5. The quantitative relevance of circulating lactate or other metabolic intermediates as fuels remains unclear. Here we systematically examine the fluxes of circulating metabolites in mice, and find that lactate can be a primary source of carbon for the TCA cycle and thus of energy. Intravenous infusions of 13C-labelled nutrients reveal that, on a molar basis, the circulatory turnover flux of lactate is the highest of all metabolites and exceeds that of glucose by 1.1-fold in fed mice and 2.5-fold in fasting mice; lactate is made primarily from glucose but also from other sources. In both fed and fasted mice, 13C-lactate extensively labels TCA cycle intermediates in all tissues. Quantitative analysis reveals that during the fasted state, the contribution of glucose to tissue TCA metabolism is primarily indirect (via circulating lactate) in all tissues except the brain. In genetically engineered lung and pancreatic cancer tumours in fasted mice, the contribution of circulating lactate to TCA cycle intermediates exceeds that of glucose, with glutamine making a larger contribution than lactate in pancreatic cancer. Thus, glycolysis and the TCA cycle are uncoupled at the level of lactate, which is a primary circulating TCA substrate in most tissues and tumours.
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We thank C. Wright for providing the Ptf1a-cre mice; M. Sander for help and advice; and the members of the Rabinowitz laboratory and J. Baur, Z. Arany, and M. Lazar for scientific discussions. This work was supported by NIH grants 1DP1DK113643, R01 CA163591, R01 CA130893, K22 CA190521, R01 CA186043, R35 CA197699, R50 CA211437, P30 CA072720 (Metabolomics Shared Resource, Rutgers Cancer Institute of New Jersey), and 5P30 DK019525. In addition, it was supported by a Stand Up To Cancer–Cancer Research UK–Lustgarten Foundation Pancreatic Cancer Dream Team Research Grant (grant number: SU2C-AACR-DT-20-16). S.H. is a Merck Fellow of the Life Sciences Research Foundation. C.J. is a postdoctoral fellow of the American Diabetes Association.
The authors declare no competing financial interests.
Reviewer Information Nature thanks S. Kempa, M. Yuneeva and the other anonymous reviewer(s) for their contribution to the peer review of this work.
Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Extended data figures and tables
Metabolites (n = 39) with reported concentration greater than 30 μM in mouse plasma. Left bar graph shows those >100 μM (n = 17) and right bar graph those between 30 μM and 100 μM (n = 22). Most of the data are from the Mouse Multiple Tissue Metabolome Database (http://mmdb.iab.keio.ac.jp) (n = 2 mice), except for glucose (n = 6 mice), acetate (n = 3 mice), and glycerol (n = 3 mice), whose concentrations were determined in this study. The metabolites shown with red bars (n = 30) are those whose turnover fluxes have been determined (Table 1). Values are mean ± s.d. The concentration cut off of 30 μM was calculated using equation (1) where we used a cardiac output of 0.5 ml g−1 min−1 (see Supplementary Note 1 for references) and a turnover flux equating to 10% of glucose Fcirc.
a, Illustration of the mouse infusion experimental setup. b, Relative total ion counts (TICs) of serum glucose and lactate during 13C-glucose infusion (individual mouse data are shown for three mice for each condition). c, Relative TICs of serum glucose and lactate during 13C-lactate infusion. d, Glucose (n = 16 for 1× and n = 6 for 0.5×; P = 0.61) and lactate (n = 18 for 1× and n = 6 for 0.5×; P = 0.50) turnover fluxes determined using two different infusion rates (mean ± s.d.). P values were determined by a two-tailed unpaired Student’s t-test. The 1× infusion rates are listed in Supplementary Table 1.
a, The dependence of lactate turnover flux (Fcirc) on the exchanging flux (forward (Jf) and reverse (Jr)) between circulating lactate and tissue pyruvate. Rapid exchanging flux does not lead to infinitely fast lactate turnover flux. Instead, it leads to a lactate turnover flux approaching the net production rate of pyruvate (Jg), as illustrated in lower panel. Jt is the pyruvate flux going to the TCA cycle. See Supplementary Note 2 for derivation. b, Spatial dependence of tracer enrichment. Labelling (L(x)) decreases in an exponential manner across a tissue capillary bed (shown schematically in the shading of the cylinder representing tissue) with the extent of arteriovenous difference in tracer labelling depending on the metabolic transformation rate (k) relative to the volumetric blood flow rate (q = Q/V; V is tissue volume) as shown in the equation. La and Lv are labelled fraction in the artery and the vein, respectively. See Supplementary Note 1. c, Lactate labelled fraction in arterial (carotid artery; n = 8 mice, mean ± s.d.) and venous (tail vein and vena cava; n = 3 mice, mean ± s.d.) serum samples, and in tail snip serum sample (n = 3 mice, mean ± s.d.). For comparison of tail snip to vena cava only, samples were collected under anaesthesia to allow access to the inferior vena cava. The difference in lactate labelling between the carotid artery and tail snip can be used to calculate lactate Fcirc using equation (3). With Q = 0.53 ± 0.11 ml min−1 g−1 and C = 2.5 ± 0.2 mM, together with , we get lactate Fcirc as 398 ± 88 nmol min−1 g−1 in the fasted state, which is comparable to the value of 374 ± 112 nmol min−1 g−1 obtained with equation (2). See Supplementary Note 1 for details.
Extended Data Figure 4 Isotopic labelling of tissue TCA intermediates reaches steady state after 2.5-h infusion of 13C-glucose.
a, Comparison of normalized labelling of tissue malate after 2.5 h (n = 5 mice; mean ± s.d.) and after 5 h of [U-13C]glucose infusion (n = 3 mice; mean ± s.d.). P values were determined by an unpaired Student’s t-test, corrected for multiple comparisons using the Holm–Sidak method. Normalized labelling is the fraction of 13C atoms in a metabolite divided by the fraction of 13C atoms in serum glucose. None of the differences are significant (P > 0.14 for the brain and liver, and P > 0.98 for other tissues). b, Comparison of normalized labelling of tissue succinate after 2.5 h (n = 5 mice; mean ± s.d.) and after 5 h of [U-13C]glucose infusion (n = 3 mice; mean ± s.d.). None of the differences are significant (P > 0.18 for the brain and liver, and P > 0.85 for other tissues).
Extended Data Figure 5 Isotope labelling of central carbon metabolites by 13C-glucose and 13C-lactate.
a, Normalized labelling by 13C-glucose in fasting mice (n = 3 for pyruvate, 2-oxoglutarate, and 3-phosphoglycerate, n = 4 for alanine, and n = 5 for all other metabolites; mean ± s.d.). b, Normalized labelling by 13C-lactate in fasting mice (n = 3 for 3-phosphoglycerate and n = 4 for all other metabolites; mean ± s.d.). n indicates the number of mice. For the lactate tracer studies, venous serum and tissue labelling are normalized to the arterial serum lactate labelling. Note that citrate, malate and succinate in tissues turn over sufficiently slowly that labelling is robust to the small (<90 s) delay between euthanizing the mouse and tissue harvesting. This delay may, however, result in erroneous measurements for tissue lactate, pyruvate and glycolytic intermediates. Analyses in the main text are limited to the better validated measurements of serum metabolites and tissue TCA intermediates. With this caveat in mind, it is nevertheless intriguing that lactate labelling varies markedly across tissues. After labelled glucose infusion, lactate labelling is highest in the brain, consistent with its use of glucose as a major substrate. In the kidneys lactate is strongly labelled after glucose infusion, even though TCA intermediates are more labelled after lactate infusion. A potential explanation involves tissue heterogeneity; for example, the presence both of glycolytic cells that make lactate from circulating glucose and of oxidative cells that make TCA intermediates from circulating lactate. In other tissues, such as liver, tissue lactate labelling is far below circulating lactate and very similar to TCA labelling; this may reflect mixing of carbon between lactate and TCA intermediates via gluconeogenesis or pyruvate cycling. Another factor diluting tissue lactate labelling is that, as blood passes through tissue, owing to the rapid exchange between tissue and circulating lactate, the circulating lactate loses its labelling, as is evident from the lactate arteriovenous labelling difference.
Extended Data Figure 6 Concentrations of succinate, malate, and citrate in mouse plasma and tissues.
Unlike citrate, succinate and malate have substantially higher concentrations in tissues than in the bloodstream, thus making them a suitable readout for the tissue TCA cycle. Data are from the Mouse Multiple Tissue Metabolome Database (http://mmdb.iab.keio.ac.jp). Values are mean ± s.d. (n = 2 mice). Note that the y-axis is a logarithmic scale.
a, Turnover fluxes of glucose (n = 4 mice, mean ± s.d.) and lactate (n = 3 mice, mean ± s.d.) in anaesthetized mice. b, Normalized labelling of serum glucose, lactate, and glutamine in anaesthetized mice with 13C-glucose infusion (n = 4 mice; mean ± s.d.) and 13C-lactate infusion (n = 3 mice; mean ± s.d.). c, Steady-state whole-body flux model summarizing glucose and lactate interconversion and their feeding to the TCA (see Supplementary Note 4). Values are mean ± s.e.m.
Extended Data Figure 8 Normalized labelling of serum glutamine, glucose, and lactate, and of tissue TCA intermediates in fed mice.
a, 13C-lactate infusion (n = 5 mice). b 13C-glutamine infusion (n = 3 mice). c, 13C-glucose infusion (n = 4 mice). Bars are mean ± s.d.
Extended Data Figure 9 Scatter plots of normalized labelling of TCA intermediates in the three types of tumours by 13C-glucose versus that by 13C-lactate.
a, KrasLSL-G12D/+Trp53−/− (KP) non-small cell lung cancer (n = 3 for 13C-glucose and 13C-lactate infusions, and n = 4 for 13C-glutamine infusion). b, KrasLSL-G12D/+Stk11−/− (KL) lung cancer (n = 3 mice for infusion of each tracer). c, KrasLSL-G12D/+Trp53−/−Ptf1aCRE/+ (KPf/fC) pancreatic ductal adenocarcinoma (n = 4 for 13C-glucose infusion, n = 3 for 13C-lactate and 13C-glutamine infusions). Values are mean ± s.d. Data are from Fig. 4a–c. The solid line represents the expected labelling by 13C-glucose assuming that glucose feeds the TCA cycle solely through circulating lactate. The dashed line indicates the expected labelling by 13C-lactate, assuming that lactate feeds the TCA cycle solely through circulating glucose.
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Hui, S., Ghergurovich, J., Morscher, R. et al. Glucose feeds the TCA cycle via circulating lactate. Nature 551, 115–118 (2017). https://doi.org/10.1038/nature24057
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