Creatine metabolism: energy homeostasis, immunity and cancer biology

Abstract

Perturbations in metabolic processes are associated with diseases such as obesity, type 2 diabetes mellitus, certain infections and some cancers. A resurgence of interest in creatine biology is developing, with new insights into a diverse set of regulatory functions for creatine. This resurgence is primarily driven by technological advances in genetic engineering and metabolism as well as by the realization that this metabolite has key roles in cells beyond the muscle and brain. Herein, we highlight the latest advances in creatine biology in tissues and cell types that have historically received little attention in the field. In adipose tissue, creatine controls thermogenic respiration and loss of this metabolite impairs whole-body energy expenditure, leading to obesity. We also cover the various roles that creatine metabolism has in cancer cell survival and the function of the immune system. Renewed interest in this area has begun to showcase the therapeutic potential that lies in understanding how changes in creatine metabolism lead to metabolic disease.

Key points

  • Mitochondria in brown adipose tissue are capable of normal oxidative phosphorylation, with P:O ratios similar to those of other tissues.

  • Atypical actions of creatine involve phosphocreatine transport into colorectal cancer cells, super-stoichiometric ADP liberation to trigger respiration in thermogenic adipocytes and chromatin remodelling to modulate macrophage polarity.

  • Cyclocreatine and creatine can both inhibit tumour progression, suggesting that the pro-cancer role of creatine is independent of its function in energy buffering.

  • The mitochondrial network transduces energy over long distances, thus minimizing the requirement for metabolite diffusion, whereas cells with a disrupted mitochondrial network might buffer energy via the creatine kinase–phosphocreatine circuit.

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Fig. 1: Pathway of endogenous creatine synthesis.
Fig. 2: UCP1-dependent thermogenesis.
Fig. 3: Creatine biology in various cell types.

References

  1. 1.

    da Silva, R. P., Clow, K., Brosnan, J. T. & Brosnan, M. E. Synthesis of guanidinoacetate and creatine from amino acids by rat pancreas. Br. J. Nutr. 111, 571–577 (2014).

    PubMed  Article  CAS  Google Scholar 

  2. 2.

    Kazak, L. et al. Genetic depletion of adipocyte creatine metabolism inhibits diet-induced thermogenesis and drives obesity. Cell Metab. 26, 660–671 (2017).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  3. 3.

    Russell, A. P. et al. Creatine transporter (SLC6A8) knockout mice display an increased capacity for in vitro creatine biosynthesis in skeletal muscle. Front. Physiol. 5, 314 (2014).

    PubMed  PubMed Central  Article  Google Scholar 

  4. 4.

    Haas, R. C. & Strauss, A. W. Separate nuclear genes encode sarcomere-specific and ubiquitous human mitochondrial creatine kinase isoenzymes. J. Biol. Chem. 265, 6921–6927 (1990).

    CAS  PubMed  Google Scholar 

  5. 5.

    Hossle, J. P. et al. Distinct tissue specific mitochondrial creatine kinases from chicken brain and striated muscle with a conserved CK framework. Biochem. Biophys. Res. Commun. 151, 408–416 (1988).

    CAS  PubMed  Article  Google Scholar 

  6. 6.

    Wallimann, T., Wyss, M., Brdiczka, D., Nicolay, K. & Eppenberger, H. M. Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands: the ‘phosphocreatine circuit’ for cellular energy homeostasis. Biochem. J. 281, 21–40 (1992).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  7. 7.

    Li, H. et al. Regulation of the creatine transporter by AMP-activated protein kinase in kidney epithelial cells. Am. J. Physiol. Ren. Physiol. 299, F167–F177 (2010).

    CAS  Article  Google Scholar 

  8. 8.

    Bessman, S. P. & Geiger, P. J. Transport of energy in muscle: the phosphorylcreatine shuttle. Science 211, 448–452 (1981).

    CAS  PubMed  Article  Google Scholar 

  9. 9.

    Glancy, B. et al. Mitochondrial reticulum for cellular energy distribution in muscle. Nature 523, 617–620 (2015).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  10. 10.

    Meyer, R. A., Sweeney, H. L. & Kushmerick, M. J. A simple analysis of the ‘‘phosphocreatine shuttle’’. Am. J. Physiol. 246, C365–C377 (1984).

    CAS  PubMed  Article  Google Scholar 

  11. 11.

    Kenyon, G. L. & Reed, G. H. Creatine kinase: structure-activity relationships. Adv. Enzymol. Relat. Areas Mol. Biol. 54, 367–426 (1983).

    CAS  PubMed  Google Scholar 

  12. 12.

    Schlattner, U., Tokarska-Schlattner, M. & Wallimann, T. Mitochondrial creatine kinase in human health and disease. Biochim. Biophys. Acta 1762, 164–180 (2006).

    CAS  PubMed  Article  Google Scholar 

  13. 13.

    Bessman, S. P. The physiological significance of the creatine phosphate shuttle. Adv. Exp. Med. Biol. 194, 1–11 (1986).

    CAS  PubMed  Article  Google Scholar 

  14. 14.

    Riesberg, L. A., Weed, S. A., McDonald, T. L., Eckerson, J. M. & Drescher, K. M. Beyond muscles: the untapped potential of creatine. Int. Immunopharmacol. 37, 31–42 (2016).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  15. 15.

    Wyss, M. & Kaddurah-Daouk, R. Creatine and creatinine metabolism. Physiol. Rev. 80, 1107–1213 (2000).

    CAS  PubMed  Article  Google Scholar 

  16. 16.

    Dzeja, P. P. & Terzic, A. Phosphotransfer networks and cellular energetics. J. Expt. Biol. 206, 2039–2047 (2003).

    CAS  Article  Google Scholar 

  17. 17.

    Schlattner, U. et al. Cellular compartmentation of energy metabolism: creatine kinase microcompartments and recruitment of B-type creatine kinase to specific subcellular sites. Amino Acids 48, 1751–1774 (2016).

    CAS  PubMed  Article  Google Scholar 

  18. 18.

    Braun, K., Oeckl, J., Westermeier, J., Li, Y. & Klingenspor, M. Non-adrenergic control of lipolysis and thermogenesis in adipose tissues. J. Exp. Biol. 221 (Suppl. 1), jeb165381 (2018).

    PubMed  Article  Google Scholar 

  19. 19.

    Rosen, E. D. & Spiegelman, B. M. What we talk about when we talk about fat. Cell 156, 20–44 (2014).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  20. 20.

    Foster, D. O. & Frydman, M. L. Brown adipose tissue: the dominant site of nonshivering thermogenesis in the rat. Experientia Suppl. 32, 147–151 (1978).

    CAS  PubMed  Article  Google Scholar 

  21. 21.

    Rothwell, N. J. & Stock, M. J. A role for brown adipose tissue in diet-induced thermogenesis. Nature 281, 31–35 (1979).

    CAS  PubMed  Article  Google Scholar 

  22. 22.

    Chouchani, E. T., Kazak, L. & Spiegelman, B. M. New advances in adaptive thermogenesis: UCP1 and beyond. Cell Metab. 29, 27–37 (2019).

    CAS  PubMed  Article  Google Scholar 

  23. 23.

    Lowell, B. B. et al. Development of obesity in transgenic mice after genetic ablation of brown adipose tissue. Nature 366, 740–742 (1993).

    CAS  PubMed  Article  Google Scholar 

  24. 24.

    Cohen, P. et al. Ablation of PRDM16 and beige adipose causes metabolic dysfunction and a subcutaneous to visceral fat switch. Cell 156, 304–316 (2014).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  25. 25.

    van Marken Lichtenbelt, W. D. et al. Cold-activated brown adipose tissue in healthy men. N. Engl. J. Med. 360, 1500–1508 (2009).

    PubMed  Article  Google Scholar 

  26. 26.

    Carpentier, A. C. et al. Brown adipose tissue energy metabolism in humans. Front. Endocrinol. 9, 447 (2018).

    Article  Google Scholar 

  27. 27.

    Leitner, B. P. et al. Mapping of human brown adipose tissue in lean and obese young men. Proc. Natl Acad. Sci. USA 114, 8649–8654 (2017).

    CAS  PubMed  Article  Google Scholar 

  28. 28.

    Golozoubova, V. et al. Only UCP1 can mediate adaptive nonshivering thermogenesis in the cold. FASEB J. 15, 2048–2050 (2001).

    CAS  PubMed  Article  Google Scholar 

  29. 29.

    Meyer, C. W. et al. Adaptive thermogenesis and thermal conductance in wild-type and UCP1-KO mice. Am. J. Physiol. Regul. Integr. Comp. Physiol. 299, R1396–R1406 (2010).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  30. 30.

    Ukropec, J., Anunciado, R. P., Ravussin, Y., Hulver, M. W. & Kozak, L. P. UCP1-independent thermogenesis in white adipose tissue of cold-acclimated Ucp1 −/− mice. J. Biol. Chem. 281, 31894–31908 (2006).

    CAS  PubMed  Article  Google Scholar 

  31. 31.

    Keipert, S. et al. Long-term cold adaptation does not require FGF21 or UCP1. Cell Metab. 26, 437–446 (2017).

    CAS  PubMed  Article  Google Scholar 

  32. 32.

    Golozoubova, V., Cannon, B. & Nedergaard, J. UCP1 is essential for adaptive adrenergic nonshivering thermogenesis. Am. J. Physiol. Endocrinol. Metab. 291, E350–E357 (2006).

    CAS  PubMed  Article  Google Scholar 

  33. 33.

    Hofmann, W. E., Liu, X., Bearden, C. M., Harper, M. E. & Kozak, L. P. Effects of genetic background on thermoregulation and fatty acid-induced uncoupling of mitochondria in UCP1-deficient mice. J. Biol. Chem. 276, 12460–12465 (2001).

    CAS  PubMed  Article  Google Scholar 

  34. 34.

    Kazak, L. et al. UCP1 deficiency causes brown fat respiratory chain depletion and sensitizes mitochondria to calcium overload-induced dysfunction. Proc. Natl Acad. Sci. USA 114, 7981–7986 (2017).

    CAS  PubMed  Article  Google Scholar 

  35. 35.

    Cannon, B. & Nedergaard, J. Nonshivering thermogenesis and its adequate measurement in metabolic studies. J. Exp. Biol. 214, 242–253 (2011).

    PubMed  Article  Google Scholar 

  36. 36.

    Matthias, A. et al. Thermogenic responses in brown fat cells are fully UCP1-dependent. UCP2 or UCP3 do not substitute for UCP1 in adrenergically or fatty scid-induced thermogenesis. J. Biol. Chem. 275, 25073–25081 (2000).

    CAS  PubMed  Article  Google Scholar 

  37. 37.

    Oelkrug, R., Kutschke, M., Meyer, C. W., Heldmaier, G. & Jastroch, M. Uncoupling protein 1 decreases superoxide production in brown adipose tissue mitochondria. J. Biol. Chem. 285, 21961–21968 (2010).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  38. 38.

    Enerback, S. et al. Mice lacking mitochondrial uncoupling protein are cold-sensitive but not obese. Nature 387, 90–94 (1997).

    CAS  PubMed  Article  Google Scholar 

  39. 39.

    Rothwell, N. J. & Stock, M. J. A role for brown adipose tissue in diet-induced thermogenesis. Obes. Res. 5, 650–656 (1997).

    CAS  PubMed  Article  Google Scholar 

  40. 40.

    Feldmann, H. M., Golozoubova, V., Cannon, B. & Nedergaard, J. UCP1 ablation induces obesity and abolishes diet-induced thermogenesis in mice exempt from thermal stress by living at thermoneutrality. Cell Metab. 9, 203–209 (2009).

    CAS  PubMed  Article  Google Scholar 

  41. 41.

    Maurer, S. F., Fromme, T., Mocek, S., Zimmermann, A. & Klingenspor, M. Uncoupling protein 1 and the capacity for non-shivering thermogenesis are components of the glucose homeostatic system. Am. J. Physiol. Endocrinol. Metab. 318, E198–E215 (2020).

    CAS  PubMed  Article  Google Scholar 

  42. 42.

    Fromme, T. et al. Bile acid supplementation decreases body mass gain in C57BL/6J but not 129S6/SvEvTac mice without increasing energy expenditure. Sci. Rep. 9, 131 (2019).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  43. 43.

    Liu, X. et al. Paradoxical resistance to diet-induced obesity in UCP1-deficient mice. J. Clin. Invest. 111, 399–407 (2003).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  44. 44.

    Zietak, M. & Kozak, L. P. Bile acids induce uncoupling protein 1-dependent thermogenesis and stimulate energy expenditure at thermoneutrality in mice. Am. J. Physiol. Endocrinol. Metab. 310, E346–E354 (2016).

    PubMed  Article  Google Scholar 

  45. 45.

    Bachman, E. S. et al. betaAR signaling required for diet-induced thermogenesis and obesity resistance. Science 297, 843–845 (2002).

    CAS  PubMed  Article  Google Scholar 

  46. 46.

    Shi, F. & Collins, S. Second messenger signaling mechanisms of the brown adipocyte thermogenic program: an integrative perspective. Horm. Mol. Biol. Clin. Investig. https://doi.org/10.1515/hmbci-2017-0062 (2017).

  47. 47.

    Hamann, A., Flier, J. S. & Lowell, B. B. Decreased brown fat markedly enhances susceptibility to diet-induced obesity, diabetes, and hyperlipidemia. Endocrinology 137, 21–29 (1996).

    CAS  PubMed  Article  Google Scholar 

  48. 48.

    Douris, N. et al. Beta-adrenergic receptors are critical for weight loss but not for other metabolic adaptations to the consumption of a ketogenic diet in male mice. Mol. Metab. 6, 854–862 (2017).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  49. 49.

    Anunciado-Koza, R., Ukropec, J., Koza, R. A. & Kozak, L. P. Inactivation of UCP1 and the glycerol phosphate cycle synergistically increases energy expenditure to resist diet-induced obesity. J. Biol. Chem. 283, 27688–27697 (2008).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  50. 50.

    Granneman, J. G., Burnazi, M., Zhu, Z. & Schwamb, L. A. White adipose tissue contributes to UCP1-independent thermogenesis. Am. J. Physiol. Endocrinol. Metab. 285, E1230–E1236 (2003).

    CAS  PubMed  Article  Google Scholar 

  51. 51.

    Ukropec, J., Anunciado, R. V., Ravussin, Y. & Kozak, L. P. Leptin is required for uncoupling protein-1-independent thermogenesis during cold stress. Endocrinology 147, 2468–2480 (2006).

    CAS  PubMed  Article  Google Scholar 

  52. 52.

    Shabalina, I. G. et al. UCP1 in brite/beige adipose tissue mitochondria is functionally thermogenic. Cell Rep. 5, 1196–1203 (2013).

    CAS  PubMed  Article  Google Scholar 

  53. 53.

    Kazak, L. et al. A creatine-driven substrate cycle enhances energy expenditure and thermogenesis in beige fat. Cell 163, 643–655 (2015).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  54. 54.

    Antonacci, M. A. et al. Direct detection of brown adipose tissue thermogenesis in UCP1−/− mice by hyperpolarized (129)Xe MR thermometry. Sci. Rep. 9, 14865 (2019).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  55. 55.

    Kontani, Y. et al. UCP1 deficiency increases susceptibility to diet-induced obesity with age. Aging Cell 4, 147–155 (2005).

    CAS  PubMed  Article  Google Scholar 

  56. 56.

    Pfannenberg, C. et al. Impact of age on the relationships of brown adipose tissue with sex and adiposity in humans. Diabetes 59, 1789–1793 (2010).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  57. 57.

    Cypess, A. M. et al. Identification and importance of brown adipose tissue in adult humans. N. Engl. J. Med. 360, 1509–1517 (2009).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  58. 58.

    Harms, M. J. et al. Prdm16 is required for the maintenance of brown adipocyte identity and function in adult mice. Cell Metab. 19, 593–604 (2014).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  59. 59.

    Muller, S. et al. Proteomic analysis of human brown adipose tissue reveals utilization of coupled and uncoupled energy expenditure pathways. Sci. Rep. 6, 30030 (2016).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  60. 60.

    Bertholet, A. M. et al. Mitochondrial patch clamp of beige adipocytes reveals UCP1-positive and UCP1-negative cells both exhibiting futile creatine cycling. Cell Metab. 25, 811–822 (2017).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  61. 61.

    Kazak, L. et al. Ablation of adipocyte creatine transport impairs thermogenesis and causes diet-induced obesity. Nat. Metab. 1, 360–370 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  62. 62.

    Yamashita, H. et al. Increased growth of brown adipose tissue but its reduced thermogenic activity in creatine-depleted rats fed β-guanidinopropionic acid. Biochim. Biophys. Acta 1230, 69–73 (1995).

    PubMed  Article  Google Scholar 

  63. 63.

    Wakatsuki, T. et al. Thermogenic responses to high-energy phosphate contents and/or hindlimb suspension in rats. Jpn. J. Physiol. 46, 171–175 (1996).

    CAS  PubMed  Article  Google Scholar 

  64. 64.

    Svensson, P. A. et al. Gene expression in human brown adipose tissue. Int. J. Mol. Med. 27, 227–232 (2011).

    CAS  PubMed  Article  Google Scholar 

  65. 65.

    Wada, S. et al. The tumor suppressor FLCN mediates an alternate mTOR pathway to regulate browning of adipose tissue. Genes Dev. 30, 2551–2564 (2016).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  66. 66.

    Perna, M. K. et al. Creatine transporter deficiency leads to increased whole body and cellular metabolism. Amino Acids 48, 2057–2065 (2016).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  67. 67.

    Schulz, T. J. et al. Brown-fat paucity due to impaired BMP signalling induces compensatory browning of white fat. Nature 495, 379–383 (2013).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  68. 68.

    Rowland, L. A., Bal, N. C., Kozak, L. P. & Periasamy, M. Uncoupling protein 1 and sarcolipin are required to maintain optimal thermogenesis, and loss of both systems compromises survival of mice under cold stress. J. Biol. Chem. 290, 12282–12289 (2015).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  69. 69.

    Berlet, H. H., Bonsmann, I. & Birringer, H. Occurrence of free creatine, phosphocreatine and creatine phosphokinase in adipose tissue. Biochim. Biophys. Acta 437, 166–174 (1976).

    CAS  PubMed  Article  Google Scholar 

  70. 70.

    Din, M. U. et al. Postprandial oxidative metabolism of human brown fat indicates thermogenesis. Cell Metab. 28, 207–216.e3 (2018).

    Article  CAS  Google Scholar 

  71. 71.

    Jash, S., Banerjee, S., Lee, M. J., Farmer, S. R. & Puri, V. CIDEA transcriptionally regulates UCP1 for britening and thermogenesis in human fat cells. iScience 20, 73–89 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  72. 72.

    Pydi, S. P. et al. Adipocyte β-arrestin-2 is essential for maintaining whole body glucose and energy homeostasis. Nat. Commun. 10, 2936 (2019).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  73. 73.

    Wang, L. et al. Selective activation of Gs signaling in adipocytes causes striking metabolic improvements in mice. Mol. Metab. 27, 83–91 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  74. 74.

    Pierce, K. L., Premont, R. T. & Lefkowitz, R. J. Seven-transmembrane receptors. Nat. Rev. Mol. Cell Biol. 3, 639–650 (2002).

    CAS  PubMed  Article  Google Scholar 

  75. 75.

    Lee, C. C. et al. Naa10p inhibits beige adipocyte-mediated thermogenesis through N-α-acetylation of Pgc1alpha. Mol. Cell 76, 500–515.e8 (2019).

    CAS  PubMed  Article  Google Scholar 

  76. 76.

    Deshmukh, A. S. et al. Proteomics-based comparative mapping of the secretomes of human brown and white adipocytes reveals EPDR1 as a novel batokine. Cell Metab. 30, 963–975 (2019).

    CAS  PubMed  Article  Google Scholar 

  77. 77.

    Levy, S. E., Chen, Y. S., Graham, B. H. & Wallace, D. C. Expression and sequence analysis of the mouse adenine nucleotide translocase 1 and 2 genes. Gene 254, 57–66 (2000).

    CAS  PubMed  Article  Google Scholar 

  78. 78.

    Jacobus, W. E. & Lehninger, A. L. Creatine kinase of rat heart mitochondria. Coupling of creatine phosphorylation to electron transport. J. Biol. Chem. 248, 4803–4810 (1973).

    CAS  PubMed  Google Scholar 

  79. 79.

    Seo, J. B. et al. Knockdown of Ant2 reduces adipocyte Hypoxia and improves insulin resistance in obesity. Nat. Metab. 1, 86–97 (2019).

    CAS  PubMed  Article  Google Scholar 

  80. 80.

    Brand, M. D. et al. The basal proton conductance of mitochondria depends on adenine nucleotide translocase content. Biochem. J. 392, 353–362 (2005).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  81. 81.

    Bertholet, A. M. et al. H+ transport is an integral function of the mitochondrial ADP/ATP carrier. Nature 571, 515–520 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  82. 82.

    Streijger, F. et al. Mice lacking brain-type creatine kinase activity show defective thermoregulation. Physiol. Behav. 97, 76–86 (2009).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  83. 83.

    Streijger, F. et al. Structural and behavioural consequences of double deficiency for creatine kinases BCK and UbCKmit. Behav. Brain Res. 157, 219–234 (2005).

    CAS  PubMed  Article  Google Scholar 

  84. 84.

    Virtue, S. & Vidal-Puig, A. Assessment of brown adipose tissue function. Front. Physiol. 4, 128 (2013).

    PubMed  PubMed Central  Article  Google Scholar 

  85. 85.

    Wikstrom, J. D. et al. Hormone-induced mitochondrial fission is utilized by brown adipocytes as an amplification pathway for energy expenditure. EMBO J. 33, 418–436 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  86. 86.

    Friedman, J. R. et al. ER tubules mark sites of mitochondrial division. Science 334, 358–362 (2011).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  87. 87.

    Zotter, A., Bauerle, F., Dey, D., Kiss, V. & Schreiber, G. Quantifying enzyme activity in living cells. J. Biol. Chem. 292, 15838–15848 (2017).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  88. 88.

    Schlattner, U. et al. Divergent enzyme kinetics and structural properties of the two human mitochondrial creatine kinase isoenzymes. Biol. Chem. 381, 1063–1070 (2000).

    CAS  PubMed  Article  Google Scholar 

  89. 89.

    Menniti, F. S., Knoth, J., Peterson, D. S. & Diliberto, E. J. Jr. The in situ kinetics of dopamine beta-hydroxylase in bovine adrenomedullary chromaffin cells. Intravesicular compartmentation reduces apparent affinity for the cofactor ascorbate. J. Biol. Chem. 262, 7651–7657 (1987).

    CAS  PubMed  Google Scholar 

  90. 90.

    Nakae, Y. & Stoward, P. J. Kinetic parameters of lactate dehydrogenase in liver and gastrocnemius determined by three quantitative histochemical methods. J. Histochem. Cytochem. 45, 1427–1431 (1997).

    CAS  PubMed  Article  Google Scholar 

  91. 91.

    Joel, C. D., Neaves, W. B. & Rabb, J. M. Mitochondria of brown fat: oxidative phosphorylation sensitive to 2,4,-dinitrophenol. Biochem. Biophys. Res. Commun. 29, 490–495 (1967).

    CAS  PubMed  Article  Google Scholar 

  92. 92.

    Guillory, R. J. & Racker, E. Oxidative phosphorylation in brown adipose mitochondria. Biochim. Biophys. Acta 153, 490–493 (1968).

    CAS  PubMed  Article  Google Scholar 

  93. 93.

    Hittelman, K. J., Lindberg, O. & Cannon, B. Oxidative phosphorylation and compartmentation of fatty acid metabolism in brown fat mitochondria. Eur. J. Bio. 11, 183–192 (1969).

    CAS  Article  Google Scholar 

  94. 94.

    Williamson, J. R. Control of energy metabolism in hamster brown adipose tissue. J. Biol. Chem. 245, 2043–2050 (1970).

    CAS  PubMed  Google Scholar 

  95. 95.

    Prusiner, S. B., Cannon, B., Ching, T. M. & Lindberg, O. Oxidative metabolism in cells isolated from brown adipose tissue. 2. Catecholamine regulated respiratory control. Eur. J. Biochem. 7, 51–57 (1968).

    CAS  PubMed  Article  Google Scholar 

  96. 96.

    LaNoue, K. F., Koch, C. D. & Meditz, R. B. Mechanism of action of norepinephrine in hamster brown adipocytes. J. Biol. Chem. 257, 13740–13748 (1982).

    CAS  PubMed  Google Scholar 

  97. 97.

    Nedergaard, J. & Lindberg, O. Norepinephrine-stimulated fatty-acid release and oxygen consumption in isolated hamster brown-fat cells. Influence of buffers, albumin, insulin and mitochondrial inhibitors. Eur. J. Biochem. 95, 139–145 (1979).

    CAS  PubMed  Article  Google Scholar 

  98. 98.

    Mills, E. L. et al. Accumulation of succinate controls activation of adipose tissue thermogenesis. Nature 560, 102–106 (2018).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  99. 99.

    Yoneshiro, T. et al. BCAA catabolism in brown fat controls energy homeostasis through SLC25A44. Nature 572, 614–619 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  100. 100.

    Schreiber, R. et al. Cold-induced thermogenesis depends on ATGL-mediated lipolysis in cardiac muscle, but not brown adipose tissue. Cell Metab. 26, 753–763 (2017).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  101. 101.

    Shin, H. et al. Lipolysis in brown adipocytes is not essential for cold-induced thermogenesis in mice. Cell Metab. 26, 764–777 (2017).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  102. 102.

    Bal, N. C. et al. Sarcolipin is a newly identified regulator of muscle-based thermogenesis in mammals. Nat. Med. 18, 1575–1579 (2012).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  103. 103.

    MacPherson, R. E. et al. Sarcolipin knockout mice fed a high-fat diet exhibit altered indices of adipose tissue inflammation and remodeling. Obesity 24, 1499–1505 (2016).

    CAS  PubMed  Article  Google Scholar 

  104. 104.

    Song, A. et al. Low- and high-thermogenic brown adipocyte subpopulations coexist in murine adipose tissue. J. Clin. Invest. 130, 247–257 (2020).

    CAS  PubMed  Article  Google Scholar 

  105. 105.

    Goldrath, A. W. & Bevan, M. J. Selecting and maintaining a diverse T-cell repertoire. Nature 402, 255–262 (1999).

    CAS  PubMed  Article  Google Scholar 

  106. 106.

    Davis, M. M. & Bjorkman, P. J. T-cell antigen receptor genes and T-cell recognition. Nature 334, 395–402 (1988).

    CAS  PubMed  Article  Google Scholar 

  107. 107.

    Hogquist, K. A., Baldwin, T. A. & Jameson, S. C. Central tolerance: learning self-control in the thymus. Nat. Rev. Immunol. 5, 772–782 (2005).

    CAS  PubMed  Article  Google Scholar 

  108. 108.

    Zhang, Y., Li, H., Wang, X., Gao, X. & Liu, X. Regulation of T cell development and activation by creatine kinase B. PLoS One 4, e5000 (2009).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  109. 109.

    Davies, L. C., Jenkins, S. J., Allen, J. E. & Taylor, P. R. Tissue-resident macrophages. Nat. Immunol. 14, 986–995 (2013).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  110. 110.

    Gordon, S. & Martinez, F. O. Alternative activation of macrophages: mechanism and functions. Immunity 32, 593–604 (2010).

    CAS  PubMed  Article  Google Scholar 

  111. 111.

    DeChatelet, L. R., McCall, C. E. & Shirley, P. S. Creatine phosphokinase activity in rabbit alveolar macrophages. Infect. Immun. 7, 29–34 (1973).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  112. 112.

    Loike, J. D., Kozler, V. F. & Silverstein, S. C. Increased ATP and creatine phosphate turnover in phagocytosing mouse peritoneal macrophages. J. Biol. Chem. 254, 9558–9564 (1979).

    CAS  PubMed  Google Scholar 

  113. 113.

    Loike, J. D., Kozler, V. F. & Silverstein, S. C. Creatine kinase expression and creatine phosphate accumulation are developmentally regulated during differentiation of mouse and human monocytes. J. Exp. Med. 159, 746–757 (1984).

    CAS  PubMed  Article  Google Scholar 

  114. 114.

    Ji, L. et al. Slc6a8-mediated creatine uptake and accumulation reprogram macrophage polarization via regulating cytokine responses. Immunity 51, 272–284 (2019).

    CAS  PubMed  Article  Google Scholar 

  115. 115.

    Guminska, M., Ptak, W. & Zembala, M. Macrophage metabolism during phagocytosis and digestion of normal and IgG antibody-coated sheep erythrocytes. Enzyme 19, 24–37 (1975).

    CAS  PubMed  Article  Google Scholar 

  116. 116.

    Kuiper, J. W. et al. Creatine kinase-mediated ATP supply fuels actin-based events in phagocytosis. PLoS Biol. 6, e51 (2008).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  117. 117.

    Kelly, P. N. The cancer immunotherapy revolution. Science 359, 1344–1345 (2018).

    CAS  PubMed  Article  Google Scholar 

  118. 118.

    Di Biase, S. et al. Creatine uptake regulates CD8 T cell antitumor immunity. J. Exp. Med. 216, 2869–2882 (2019).

    PubMed  PubMed Central  Article  Google Scholar 

  119. 119.

    Turer, E. et al. Creatine maintains intestinal homeostasis and protects against colitis. Proc. Natl. Acad. Sci. USA 114, E1273–E1281 (2017).

    CAS  PubMed  Article  Google Scholar 

  120. 120.

    Gupta, G. P. & Massague, J. Cancer metastasis: building a framework. Cell 127, 679–695 (2006).

    CAS  PubMed  Article  Google Scholar 

  121. 121.

    Vander Heiden, M. G. & DeBerardinis, R. J. Understanding the intersections between metabolism and cancer biology. Cell 168, 657–669 (2017).

    PubMed Central  Article  CAS  Google Scholar 

  122. 122.

    Dupuy, F. et al. PDK1-dependent metabolic reprogramming dictates metastatic potential in breast cancer. Cell Metab. 22, 577–589 (2015).

    CAS  PubMed  Article  Google Scholar 

  123. 123.

    Fenouille, N. et al. The creatine kinase pathway is a metabolic vulnerability in EVI1-positive acute myeloid leukemia. Nat. Med. 23, 301–313 (2017).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  124. 124.

    Kurmi, K. et al. Tyrosine phosphorylation of mitochondrial creatine kinase 1 enhances a druggable tumor energy shuttle pathway. Cell Metab. 28, 833–847 (2018).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  125. 125.

    Lillie, J. W. et al. Cyclocreatine (1-carboxymethyl-2-iminoimidazolidine) inhibits growth of a broad spectrum of cancer cells derived from solid tumors. Cancer Res. 53, 3172–3178 (1993).

    CAS  PubMed  Google Scholar 

  126. 126.

    Miller, E. E., Evans, A. E. & Cohn, M. Inhibition of rate of tumor growth by creatine and cyclocreatine. Proc. Natl Acad. Sci. USA 90, 3304–3308 (1993).

    CAS  PubMed  Article  Google Scholar 

  127. 127.

    Annesley, T. M. & Walker, J. B. Cyclocreatine phosphate as a substitute for creatine phosphate in vertebrate tissues. Energistic considerations. Biochem. Biophys. Res. Commun. 74, 185–190 (1977).

    CAS  PubMed  Article  Google Scholar 

  128. 128.

    Kristensen, C. A., Askenasy, N., Jain, R. K. & Koretsky, A. P. Creatine and cyclocreatine treatment of human colon adenocarcinoma xenografts: 31P and 1H magnetic resonance spectroscopic studies. Br. J. Cancer 79, 278–285 (1999).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  129. 129.

    Gaidzik, V. I. et al. RUNX1 mutations in acute myeloid leukemia: results from a comprehensive genetic and clinical analysis from the AML study group. J. Clin. Oncol. 29, 1364–1372 (2011).

    PubMed  Article  Google Scholar 

  130. 130.

    Feld, R. D. & Witte, D. L. Presence of creatine kinase BB isoenzyme in some patients with prostatic carcinoma. Clin. Chem. 23, 1930–1932 (1977).

    CAS  PubMed  Article  Google Scholar 

  131. 131.

    Gazdar, A. F. et al. Levels of creatine kinase and its BB isoenzyme in lung cancer specimens and cultures. Cancer Res. 41, 2773–2777 (1981).

    CAS  PubMed  Google Scholar 

  132. 132.

    Li, X. H. et al. Knockdown of creatine kinase B inhibits ovarian cancer progression by decreasing glycolysis. Int. J. Biochem. Cell Biol. 45, 979–986 (2013).

    PubMed  Article  CAS  Google Scholar 

  133. 133.

    Glass, C., Wilson, M., Gonzalez, R., Zhang, Y. & Perkins, A. S. The role of EVI1 in myeloid malignancies. Blood Cells Mol. Dis. 53, 67–76 (2014).

    CAS  PubMed  Article  Google Scholar 

  134. 134.

    Benajiba, L. et al. Creatine kinase pathway inhibition alters GSK3 and WNT signaling in EVI1-positive AML. Leukemia 33, 800–804 (2019).

    PubMed  Article  Google Scholar 

  135. 135.

    Saks, V. A., Dzhaliashvili, I. V., Konorev, E. A. & Strumia, E. Molecular and cellular aspects of the cardioprotective mechanism of phosphocreatine. Biokhimiia 57, 1763–1784 (1992).

    CAS  PubMed  Google Scholar 

  136. 136.

    Orth, M. F. et al. Functional genomics identifies AMPD2 as a new prognostic marker for undifferentiated pleomorphic sarcoma. Int. J. Cancer 144, 859–867 (2019).

    CAS  PubMed  Article  Google Scholar 

  137. 137.

    Wong, M. et al. AMPD3 is associated with the malignant characteristics of gastrointestinal stromal tumors. Oncol. Lett. 13, 1281–1287 (2017).

    CAS  PubMed  Article  Google Scholar 

  138. 138.

    Tokarska-Schlattner, M. et al. Phosphocreatine interacts with phospholipids, affects membrane properties and exerts membrane-protective effects. PLoS One 7, e43178 (2012).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  139. 139.

    Zucchi, R. et al. Protection of isolated rat heart from oxidative stress by exogenous creatine phosphate. J. Mol. Cell. Cardiol. 21, 67–73 (1989).

    CAS  PubMed  Article  Google Scholar 

  140. 140.

    Johannsen, S. et al. Screening test for malignant hyperthermia in patients with persistent hyperCKemia: a pilot study. Muscle Nerve 47, 677–681 (2013).

    CAS  PubMed  Article  Google Scholar 

  141. 141.

    Loo, J. M. et al. Extracellular metabolic energetics can promote cancer progression. Cell 160, 393–406 (2015).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  142. 142.

    Ackerman, S. E., Blackburn, O. A., Marchildon, F. & Cohen, P. Insights into the link between obesity and cancer. Curr. Obes. Rep. 6, 195–203 (2017).

    PubMed  Article  Google Scholar 

  143. 143.

    Olson, O. C., Quail, D. F. & Joyce, J. A. Obesity and the tumor microenvironment. Science 358, 1130–1131 (2017).

    CAS  PubMed  Article  Google Scholar 

  144. 144.

    Park, J. & Scherer, P. E. Endotrophin - a novel factor linking obesity with aggressive tumor growth. Oncotarget 3, 1487–1488 (2012).

    PubMed  PubMed Central  Article  Google Scholar 

  145. 145.

    Khandekar, M. J., Cohen, P. & Spiegelman, B. M. Molecular mechanisms of cancer development in obesity. Nat. Rev. Cancer 11, 886–895 (2011).

    CAS  PubMed  Article  Google Scholar 

  146. 146.

    Hamady, Z. Z. et al. Fatty liver disease as a predictor of local recurrence following resection of colorectal liver metastases. Br. J. Surg. 100, 820–826 (2013).

    CAS  PubMed  Article  Google Scholar 

  147. 147.

    VanSaun, M. N., Lee, I. K., Washington, M. K., Matrisian, L. & Gorden, D. L. High fat diet induced hepatic steatosis establishes a permissive microenvironment for colorectal metastases and promotes primary dysplasia in a murine model. Am. J. Pathol. 175, 355–364 (2009).

    PubMed  PubMed Central  Article  Google Scholar 

  148. 148.

    Ohashi, K. et al. NOD-like receptor C4 inflammasome regulates the growth of colon cancer liver metastasis in NAFLD. Hepatology 70, 1582–1599 (2019).

    CAS  PubMed  Article  Google Scholar 

  149. 149.

    Wu, Y. et al. Insulin-like growth factor-I regulates the liver microenvironment in obese mice and promotes liver metastasis. Cancer Res. 70, 57–67 (2010).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  150. 150.

    Smith, B. K. et al. Treatment of nonalcoholic fatty liver disease: role of AMPK. Am. J. Physiol. Endocrinol. Metab. 311, E730–E740 (2016).

    PubMed  Article  Google Scholar 

  151. 151.

    Komatsu, M. et al. NNMT activation can contribute to the development of fatty liver disease by modulating the NAD+ metabolism. Sci Rep. 8, 8637 (2018).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  152. 152.

    Lowe, M. T., Kim, E. H., Faull, R. L., Christie, D. L. & Waldvogel, H. J. Dissociated expression of mitochondrial and cytosolic creatine kinases in the human brain: a new perspective on the role of creatine in brain energy metabolism. J. Cereb. Blood Flow Metab. 33, 1295–1306 (2013).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  153. 153.

    Kuiper, J. W. et al. Local ATP generation by brain-type creatine kinase (CK-B) facilitates cell motility. PLoS One 4, e5030 (2009).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  154. 154.

    Nevo, A. C. & Rikmenspoel, R. Diffusion of ATP in sperm flagella. J. Theor. Biol. 26, 11–18 (1970).

    CAS  PubMed  Article  Google Scholar 

  155. 155.

    Tombes, R. M., Brokaw, C. J. & Shapiro, B. M. Creatine kinase-dependent energy transport in sea urchin spermatozoa. Flagellar wave attenuation and theoretical analysis of high energy phosphate diffusion. Biophys. J. 52, 75–86 (1987).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  156. 156.

    Papalazarou, V. et al. The creatine–phosphagen system is mechanoresponsive in pancreatic adenocarcinoma and fuels invasion and metastasis. Nat. Metab. 2, 62–80 (2020).

    CAS  Article  Google Scholar 

  157. 157.

    Ellington, W. R. Evolution and physiological roles of phosphagen systems. Annu. Rev. Physiol. 63, 289–325 (2001).

    CAS  PubMed  Article  Google Scholar 

  158. 158.

    Schwarz, T. L. Mitochondrial trafficking in neurons. Cold Spring Harb. Perspect. Biol. 5, a011304 (2013).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  159. 159.

    Kaldis, P. et al. Identification of two distinctly localized mitochondrial creatine kinase isoenzymes in spermatozoa. J. Cell Sci. 109, 2079–2088 (1996).

    CAS  PubMed  Google Scholar 

  160. 160.

    Skulachev, V. P. Mitochondrial filaments and clusters as intracellular power-transmitting cables. Trends Biochem. Sci. 26, 23–29 (2001).

    CAS  PubMed  Article  Google Scholar 

  161. 161.

    Steeghs, K. et al. Cytoarchitectural and metabolic adaptations in muscles with mitochondrial and cytosolic creatine kinase deficiencies. Mol. Cell. Biochem. 184, 183–194 (1998).

    CAS  PubMed  Article  Google Scholar 

  162. 162.

    van Deursen, J. et al. Skeletal muscles of mice deficient in muscle creatine kinase lack burst activity. Cell 74, 621–631 (1993).

    PubMed  Article  Google Scholar 

  163. 163.

    van Deursen, J. et al. Creatine kinase (CK) in skeletal muscle energy metabolism: a study of mouse mutants with graded reduction in muscle CK expression. Proc. Natl Acad. Sci. USA 91, 9091–9095 (1994).

    PubMed  Article  Google Scholar 

  164. 164.

    Steeghs, K. et al. Use of gene targeting for compromising energy homeostasis in neuro-muscular tissues: the role of sarcomeric mitochondrial creatine kinase. J. Neurosci. Methods 71, 29–41 (1997).

    CAS  PubMed  Article  Google Scholar 

  165. 165.

    Steeghs, K. et al. Mouse ubiquitous mitochondrial creatine kinase: gene organization and consequences from inactivation in mouse embryonic stem cells. DNA Cell Biol. 14, 539–553 (1995).

    CAS  PubMed  Article  Google Scholar 

  166. 166.

    Ligon, L. A. & Steward, O. Role of microtubules and actin filaments in the movement of mitochondria in the axons and dendrites of cultured hippocampal neurons. J. Comp. Neurol. 427, 351–361 (2000).

    CAS  PubMed  Article  Google Scholar 

  167. 167.

    Campello, S. et al. Orchestration of lymphocyte chemotaxis by mitochondrial dynamics. J. Exp. Med. 203, 2879–2886 (2006).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  168. 168.

    Mills, K. M., Brocardo, M. G. & Henderson, B. R. APC binds the miro/milton motor complex to stimulate transport of mitochondria to the plasma membrane. Mol. Biol. Cell 27, 466–482 (2016).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  169. 169.

    Ferrante, A. W. Jr. The immune cells in adipose tissue. Diabetes Obes. Metab. 15 (Suppl. 3), 34–38 (2013).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  170. 170.

    Lumeng, C. N., Bodzin, J. L. & Saltiel, A. R. Obesity induces a phenotypic switch in adipose tissue macrophage polarization. J. Clin. Invest. 117, 175–184 (2007).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  171. 171.

    Lumeng, C. N., Deyoung, S. M., Bodzin, J. L. & Saltiel, A. R. Increased inflammatory properties of adipose tissue macrophages recruited during diet-induced obesity. Diabetes 56, 16–23 (2007).

    CAS  PubMed  Article  Google Scholar 

  172. 172.

    Guilherme, A., Virbasius, J. V., Puri, V. & Czech, M. P. Adipocyte dysfunctions linking obesity to insulin resistance and type 2 diabetes. Nat. Rev. Mol. Cell Biol. 9, 367–377 (2008).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  173. 173.

    Kanneganti, T. D. & Dixit, V. D. Immunological complications of obesity. Nat. Immunol. 13, 707–712 (2012).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

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Acknowledgements

The authors apologize for being unable to cite papers that have contributed to the progress of this field owing to space limitations. The authors acknowledge support by the Canadian Institutes of Health Research (CIHR; grant PJT-159529) and Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant (to L.K.).

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Glossary

Creatine kinase–phosphocreatine circuit

Also known as the phosphocreatine shuttle, this system mediates the stoichiometric (1:1) transphosphorylation of phosphate from mitochondrial or glycolytic ATP to phosphocreatine, which is then used by creatine kinase to maintain high local ATP:ADP ratios.

UCP1

A mitochondrial inner membrane protein that dissipates the proton gradient across the lipid bilayer, effectively decreasing the proton-motive force and minimizing ATP synthesis; the energy dissipated across the mitochondrial inner membrane results in a considerable increase in the rate of respiration, substrate oxidation and release of heat.

Proton-motive force

The potential energy stored as a combination of the electrical and concentration (electrochemical) gradient across the mitochondrial inner membrane due to the extrusion of protons into the intermembrane space by the electron transport chain.

Congenic background

An inbred strain of mouse where the control and experimental animals only differ from one another by a small genetic region (typically a single gene).

Thermoneutrality

The ambient temperature where the metabolic rate is at a minimum, when temperature regulation is achieved by non-evaporative physical processes alone.

Creatine-dependent thermogenesis

The phosphorylation of creatine by creatine kinase and subsequent dephosphorylation of phosphocreatine (or downstream phosphometabolite) that regenerates creatine and dissipates the high-energy phosphate to generate heat; also known as futile creatine cycling.

P:O ratio

The number of moles of ADP phosphorylated to ATP for every two electrons that reduce oxygen to water.

Thymocyte selection

During T cell differentiation, thymocytes can undergo expansion, differentiation (positive selection) or cell death (negative selection).

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Kazak, L., Cohen, P. Creatine metabolism: energy homeostasis, immunity and cancer biology. Nat Rev Endocrinol (2020). https://doi.org/10.1038/s41574-020-0365-5

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