Experimental evidence on the efficacy of two new metabolic modulators on mitochondrial biogenesis and function in mouse cardiomyocytes

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Laura Tedesco
Fabio Rossi
Chiara Ruocco
Maurizio Ragni
Michele O. Carruba
Alessandra Valerio
Enzo Nisoli


branched-chain amino acids; essential amino acids; mitochondria; cardiomyocytes; tricarboxylic acid cycle


Proper maintenance of mitochondrial homeostasis is essential for cell health, and mitochondrial dysfunction underlies several metabolic and heart diseases. Stimulation of mitochondrial biogenesis represents a valuable therapeutic tool for the prevention and treatment of disorders characterized by a deficit in energy metabolism. The present study aimed to potentiate the mitochondrial biogenetic efficacy of an amino acid (AA) mixture, enriched in branched-chain amino acids (BCAAs), which we previously showed to boost mitochondrial biogenesis, leading to life span extension and reducing of muscle and liver damage. Hence, we designed and studied several innovative mixtures. Here, we report on two new AA formulas, ?5 and E7, created on the BCAA-enriched amino acid mixture (BCAAem) template and enriched with Krebs cycle substrates, including succinate, malate, and citrate. Cardiomyocytes in culture exposed to either mixture showed increased mitochondrial DNA amount, mitochondrial biogenesis markers, and oxygen consump-tion. Furthermore, ?5 and E7 also increased the expression of BCAA catabolic genes. Most importantly, all of these effects of ?5 and E7 were more pronounced than those observed with BCAAem, confirming the higher mitochondrial biogenesis potential of these new formulas. Therefore, ?5 and E7 could represent a more efficient tool for the nutritional treatment of diseases in which energy production is defective.

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1. Wlodek D, Gonzales M. Decreased energy levels can cause and sustain obesity. J Theor Biol 2003;225:33–44. http://dx.doi.org/10.1016/S0022-5193(03)00218-2
2. Boudina S, Sena S, O’Neill BT, et al. Reduced mitochondrial oxidative capacity and increased mitochondrial uncoupling impair myocardial energetics in obesity. Circulation 2005;112:2686–95. http://dx.doi.org/10.1161/CIRCULATIONAHA. 105.554360
3. Sparks LM, Xie H, Koza RA, et al. A high-fat diet coordinately downregulates genes required for mitochondrial oxidative phosphorylation in skeletal muscle. Diabetes 2005;54:1926–33. http://dx.doi. org/10.2337/diabetes.54.7.1926
4. Wisloff U, Najjar SM, Ellingsen O, et al. Cardiovascular risk factors emerge after artificial selection for low aerobic capacity. Science 2005;307:418–20. http://dx.doi.org/10.1126/science. 1108177
5. Zhou B, Tian R. Mitochondrial dysfunction in pathophysiology of heart failure. J Clin Invest 2018;128(9): 3716–26. http://dx.doi.org/10.1172/JCI120849
6. Brown DA, Perry JB, Allen ME, et al. Expert consensus document: Mitochondrial function as a therapeutic target in heart failure. Nat Rev Cardiol 2017;14(4):238– 50. http://dx.doi.org/10.1038/nrcardio.2016.203
7. Guarente L. Mitochondria—A nexus for aging, calorie restriction, and sirtuins? Cell 2008;132:171–6. http://dx.doi.org/10.1016/j.cell.2008.01.007
8. Nisoli E, Tonello C, Cardile A, et al. Calorie restriction promotes mitochondrial biogenesis by inducing the expression of eNOS. Science 2005;310:314–17. http://dx.doi.org/10.1126/science.1117728
9. Alvers AL, Fishwick LK, Wood MS, et al. Autophagy and amino acid homeostasis are required for chronological longevity in Saccharomyces cerevisiae. Aging Cell 2009;8:353–69. http://dx.doi. org/10.1111/j.1474-9726.2009.00469.x
10. D’Antona G, Ragni M, Cardile A, et al. Branched-chain amino acid supplementation promotes survival and supports cardiac and skeletal muscle mitochondrial biogenesis in middle-aged mice. Cell Metab 2010;12:362–72. http://dx.doi.org/10.1016/j. cmet.2010.08.016
11. D’Antona G, Tedesco L, Ruocco C, et al. A Peculiar formula of essential amino acids prevents rosuvastatin myopathy in mice. Antioxid Redox Signal 2016;25:595–608. http://dx.doi.org/10.1089/ ars.2015.6582
12. Tedesco L, Corsetti G, Ruocco C, et al. A specific amino acid formula prevents alcoholic liver disease in rodents. Am J Physiol Gastrointest Liver Physiol 2018; 314:G566–82. http://dx.doi.org/10.1152/ ajpgi.00231.2017
13. Claycomb WC, Lanson NA Jr, Stallworth BS, et al. HL-1 cells: A cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult car-diomyocyte. Proc Natl Acad Sci U S A 1998;95:2979– 84. http://dx.doi.org/10.1073/pnas.95.6.2979
14. Wu M, Neilson A, Swift AL, et al. Multiparameter metabolic analysis reveals a close link between attenuated mitochondrial bioenergetic function and enhanced glycolysis dependency in human tumor cells. Am J Physiol Cell Physiol 2007;292(1):C125– 36. http://dx.doi.org/10.1152/ajpcell.00247.2006
15. Neinast M, Murashige D, Arany Z. Branched chain amino acids. Annu Rev Physiol 2019;81: 139–64. http://dx.doi.org/10.1146/annurev-physiol-020518-114455
16. Sun H, Olson KC, Gao C, et al. Catabolic defect of branched-chain amino acids promotes heart failure. Circulation 2016;133(21):2038–49. http://dx.doi. org/10.1161/CIRCULATIONAHA.115.020226
17. Nisoli E, Clementi E, Paolucci C, et al. Mitochondrial biogenesis in mammals: The role of endogenous nitric oxide. Science 2003;299(5608):896–9. http:// dx.doi.org/10.1126/science.1079368
18. Duan Y, Zeng L, Li F, et al. Effect of branched-chain amino acid ratio on the proliferation, differentiation, and expression levels of key regulators involved in protein metabolism of myocytes. Nutrition 2017;36:8– 16. http://dx.doi.org/10.1016/j.nut.2016.10.016
19. Wolfe RR. Branched-chain amino acids and muscle protein synthesis in humans: Myth or reality? J Int Soc Sports Nutr 2017;14:30. http://dx.doi. org/10.1186/s12970-017-0184-9
20. Solerte SB, Gazzaruso C, Bonacasa R, et al. Nutritional supplements with oral amino acid mixtures increases whole-body lean mass and insulin sensitivity in elderly subjects with sarcopenia. Am J Cardiol 2008;101:69E–77E. http://dx.doi. org/10.1016/j.amjcard.2008.03.004
21. Aquilani R, Viglio S, Iadarola P, et al. Oral amino acid supplements improve exercise capacities in elderly patients with chronic heart failure. Am J Cardiol 2008;101:104E–10E. http://dx.doi. org/10.1016/j.amjcard.2008.03.008
22. Aquilani R, Zuccarelli GC, Dioguardi FS, et al. Effects of oral amino acid supplementation on long-term-care-acquired infections in elderly patients. Arch Gerontol Geriatr 2011;52:e123–8. http://dx. doi.org/10.1016/j.archger.2010.09.005
23. Bolasco P, Caria S, Cupisti A, et al. A novel amino acids oral supplementation in hemodialysis patients: A pilot study. Ren Fail 2011;33:1–5. http://dx.doi.org /10.3109/0886022X.2010.536289
24. Dal Negro RW, Aquilani R, Bertacco S, et al. Comprehensive effects of supplemented essential amino acids in patients with severe copd and sarcopenia. Monaldi Arch Chest Dis 2010;73:25–33. http://dx.doi.org/10.4081/monaldi.2010.310
25. Buondonno I, Sassi F, Carignano G, et al. From mitochondria to healthy aging: The role of branched-chain amino acids treatment: MATeR a randomized study. Clin Nutr 2020;39(7):2080–2091. http://dx. doi.org/10.1016/j.clnu.2019.10.013
26. Owen OE, Kalhan SC, Hanson RW. The key role of anaplerosis and cataplerosis for citric acid cycle function. J Biol Chem 2002;277(34):30409–12. http://dx.doi.org/10.1074/jbc.R200006200
27. Mills EL, Pierce KA, Jedrychowski MP, et al. Accumulation of succinate controls activation of adipose tissue thermogenesis. Nature 2018; 560(7716):102–6. http://dx.doi.org/10.1038/s41586-018-0353-2
28. Yoneshiro T, Wang Q, Tajima K, et al. BCAA catabolism in brown fat controls energy homeostasis through SLC25A44. Nature 2019;572(7771):614– 619. http://dx.doi.org/10.1038/s41586-019-1503-x