European Journal of Nutrition

, Volume 55, Issue 5, pp 1863–1873 | Cite as

Prospective study of telomere length and LINE-1 methylation in peripheral blood cells: the role of B vitamins supplementation

  • Irene Pusceddu
  • Markus Herrmann
  • Susanne H. Kirsch
  • Christian Werner
  • Ulrich Hübner
  • Marion Bodis
  • Ulrich Laufs
  • Stefan Wagenpfeil
  • Jürgen Geisel
  • Wolfgang Herrmann
Original Contribution



Deficiencies of folate, vitamins B12 and D are common age-related conditions. Vitamin B12 and folate are necessary for DNA methylation. Telomeres appear to be regulated by DNA methylation. Here, we study the effect of B vitamins supplementation on telomere length and global DNA methylation in a prospective study.


In total, 60 elderly subjects were supplemented for 1 year with either vitamin B12, B6, folate, vitamin D and calcium (group A n = 31) or only vitamin D and calcium (group B n = 29). LINE-1 methylation, relative telomere length (T/S), vitamin B12, folate, homocysteine (tHcy) , 5-methyltetrahydrofolate (5-methylTHF), S-adenosylhomocysteine (SAH), S-adenosylmethionine (SAM), cystathionine and vitamin D were quantified before and after supplementation.


At baseline, tHcy was high, vitamin D was low, and T/S did not differ between groups A and B. Vitamin supplementation increased LINE-1 methylation in group A at site 317 but reduced LINE-1 methylation in group B at site 327. There was no correlation between T/S and LINE-1 methylation at baseline. Multiple backward regression analysis revealed baseline tHcy and 5-methylTHF are significant predictors of T/S. After supplementation in group B but not in group A, LINE-1 methylation correlated inversely with T/S, and LINE-1 methylation variation was an independent predictor of T/S variation. B vitamins decreased tHcy significantly in group A. Multiple backward regression analysis showed 5-methylTHF in group A and tHcy in group B were significant predictors for LINE-1 methylation. At baseline, the lower LINE-1 methylation observed in subjects with 5-methylTHF >10 nmol/l was in agreement with a reduced methyl group transfer due to a lower SAM formation. In group B, an increase in telomere length was correlated with lower LINE-1 methylation. Subjects with hyperhomocysteinemia >12 µmol/L had compared to those with normal tHcy a reduced LINE-1 methylation accompanied by a higher SAM and SAH (that inhibits demethylation of SAM) as well as lower 5-methylTHF. Additionally, subjects with tHcy > 12 µmol/L had longer telomeres when compared with subjects having tHcy < 12 µmol/L.


The results suggest a possible effect of B vitamins for telomere biology in blood cells. Suboptimal B vitamins status and hyperhomocysteinemia are associated with altered DNA methylation and telomere length. These data have to be confirmed in future studies.


B vitamins Telomere length DNA methylation 


Compliance with ethical standards

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.


  1. 1.
    Stabler SP (2013) Clinical practice. Vitamin B12 deficiency. N Engl J Med 368:149–160CrossRefGoogle Scholar
  2. 2.
    Holick MF (2007) Vitamin D deficiency. N Engl J Med 357:266–281CrossRefGoogle Scholar
  3. 3.
    Herrmann W, Schorr H, Bodis M, Knapp JP, Muller A, Stein G, Geisel J (2000) Role of homocysteine, cystathionine and methylmalonic acid measurement for diagnosis of vitamin deficiency in high-aged subjects. Eur J Clin Invest 30:1083–1089CrossRefGoogle Scholar
  4. 4.
    Hughes CF, Ward M, Hoey L, McNulty H (2013) Vitamin B12 and ageing: current issues and interaction with folate. Ann Clin Biochem 50:315–329CrossRefGoogle Scholar
  5. 5.
    Moores CJ, Fenech M, O’Callaghan NJ (2014) Telomere dynamics: the influence of folate and DNA methylation. Ann N Y Acad Sci 1229:76–88CrossRefGoogle Scholar
  6. 6.
    Wainfan E, Poirier LA (1992) Methyl groups in carcinogenesis: effects on DNA methylation and gene expression. Cancer Res 52:2071s–2077sGoogle Scholar
  7. 7.
    Nazki FHSAGB (2014) Folate: metabolism, genes, polymorphisms and the associated diseases. Gene 533:11–20CrossRefGoogle Scholar
  8. 8.
    Kazazian HH (2014) Mobile elements: drivers of genome evolution. Science 303:1626–1632CrossRefGoogle Scholar
  9. 9.
    Blasco MA (2007) Telomere length, stem cells and aging. Nat Chem Biol 3:640–649CrossRefGoogle Scholar
  10. 10.
    Blasco MA (2005) Telomeres and human disease: ageing, cancer and beyond. Nat Rev Genet 6:611–622CrossRefGoogle Scholar
  11. 11.
    Hayflick L (1965) The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 37:614–636CrossRefGoogle Scholar
  12. 12.
    Fyhrquist F, Saijonmaa O, Strandberg T (2013) The roles of senescence and telomere shortening in cardiovascular disease. Nat Rev Cardiol 10:274–283CrossRefGoogle Scholar
  13. 13.
    Cawthon RM, Smith KR, O’Brien E, Sivatchenko A, Kerber RA (2003) Association between telomere length in blood and mortality in people aged 60 years or older. Lancet 361:393–395CrossRefGoogle Scholar
  14. 14.
    Armanios M (2013) Telomeres and age-related disease: how telomere biology informs clinical paradigms. J Clin Invest 123:996–1002CrossRefGoogle Scholar
  15. 15.
    Svenson U, Nordfjall K, Stegmayr B, Manjer J, Nilsson P, Tavelin B, Henriksson R, Lenner P, Roos G (2008) Breast cancer survival is associated with telomere length in peripheral blood cells. Cancer Res 68:3618–3623CrossRefGoogle Scholar
  16. 16.
    Li H, Engström K, Vahter M, Broberg K (2014) Arsenic exposure through drinking water is associated with longer telomeres in peripheral blood. Chem Res Toxicol 25:2333–2339CrossRefGoogle Scholar
  17. 17.
    Richards JB, Valdes AM, Gardner JP, Kato BS, Siva A, Kimura M, Lu X, Brown MJ, Aviv A, Spector TD (2008) Homocysteine levels and leukocyte telomere length. Atherosclerosis 200:271–277CrossRefGoogle Scholar
  18. 18.
    Paul L, Cattaneo M, D’Angelo A, Sampietro F, Fermo I, Razzari C, Fontana G, Eugene N, Jacques PF, Selhub J (2009) Telomere length in peripheral blood mononuclear cells is associated with folate status in men. J Nutr 139:1273–1278CrossRefGoogle Scholar
  19. 19.
    Liu JJ, Prescott J, Giovannucci E, Hankinson SE, Rosner B, De Vivo I (2013) One-carbon metabolism factors and leukocyte telomere length. Am J Clin Nutr 97:794–799CrossRefGoogle Scholar
  20. 20.
    Paul L, Jacques PF, Aviv A, Vasan RS, D’Agostino RB, Levy D, Selhub J (2014) High plasma folate is negatively associated with leukocyte telomere length in the Framingham Offspring cohort. Eur J Nutr 54:235–241CrossRefGoogle Scholar
  21. 21.
    Herrmann W, Kirsch SH, Kruse V, Eckert R, Graber S, Geisel J, Obeid R (2013) One year B and D vitamins supplementation improves metabolic bone markers. Clin Chem Lab Med 51:639–647Google Scholar
  22. 22.
    Hubner U, Geisel J, Kirsch SH, Kruse V, Bodis M, Klein C, Herrmann W, Obeid R (2013) Effect of 1 year B and D vitamin supplementation on LINE-1 repetitive element methylation in older subjects. Clin Chem Lab Med 51:649–655CrossRefGoogle Scholar
  23. 23.
    Geisel J, Schorr H, Heine GH, Bodis M, Hubner U, Knapp JP, Herrmann W (2007) Decreased p66Shc promoter methylation in patients with end-stage renal disease. Clin Chem Lab Med 45:1764–1770CrossRefGoogle Scholar
  24. 24.
    Cawthon RM (2002) Telomere measurement by quantitative PCR. Nucleic Acids Res 30:2–6CrossRefGoogle Scholar
  25. 25.
    Werner C, Fürster T, Widmann T, Pöss J, Roggia C, Hanhoun M, Scharhag J, Büchner N, Meyer T, Kindermann W, Haendeler J, Böhm M, Laufs U (2009) Physical exercise prevents cellular senescence in circulating leukocytes and in the vessel wall. Circulation 120:2438–2447CrossRefGoogle Scholar
  26. 26.
    Kirsch SH, Herrmann W, Kruse V, Eckert R, Gräber S, Geisel J, Obeid R (2014) One year B-vitamins increases serum and whole blood forms and lowers plasma homocysteine in older Germans. Clin Chem Lab Med 53:445–452Google Scholar
  27. 27.
    Kirsch SH, Knapp JP, Herrmann W, Obeid R (2010) Quantification of key folate forms in serum using stable-isotope dilution ultra performance liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 878:68–75CrossRefGoogle Scholar
  28. 28.
    Stabler SP, Marcell PD, Podell ER, Allen RH (1987) Quantitation of total homocysteine, total cysteine, and methionine in normal serum and urine using capillary gas chromatography-mass spectrometry. Anal Biochem 162:185–196CrossRefGoogle Scholar
  29. 29.
    Kirsch SH, Knapp JP, Geisel J, Herrmann W, Obeid R (2009) Simultaneous quantification of S-adenosyl methionine and S-adenosyl homocysteine in human plasma by stable-isotope dilution ultra performance liquid chromatography tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 877:3865–3870CrossRefGoogle Scholar
  30. 30.
    Müezzinler A, Zaineddin AK, Brenner H (2013) A systematic review of leukocyte telomere length and age in adults. Ageing Res Rev 12:509–519CrossRefGoogle Scholar
  31. 31.
    Weischer M, Bojesen SE, Nordestgaard BG (2014) Telomere shortening unrelated to smoking, body weight, physical activity, and alcohol intake: 4576 general population individuals with repeat measurements 10 years apart. PLoS Genet 10:1–11CrossRefGoogle Scholar
  32. 32.
    Ehrlenbach S, Willeit P, Kiechl S, Willeit J, Reindl M, Schanda K, Kronenberg F, Brandstatter A (2009) Influences on the reduction of relative telomere length over 10 years in the population-based Bruneck study: introduction of a well-controlled high-throughput assay. Int J Epidemiol 38:1725–1734CrossRefGoogle Scholar
  33. 33.
    Epel ES, Merkin SS, Cawthon RM, Blackburn EH, Adler NE, Pletcher MJ, Seeman TE (2008) The rate of leukocyte telomere shortening predicts mortality from cardiovascular disease in elderly men. Aging 4:88Google Scholar
  34. 34.
    Nordfjall K, Svenson U, Norrback KF, Adolfsson R, Lenner P, Roos G (2009) The individual blood cell telomere attrition rate is telomere length dependent. PLoS Genet 5:e1000375CrossRefGoogle Scholar
  35. 35.
    Aviv A, Chen W, Gardner JP, Kimura M, Brimacombe M, Cao X, Srinivasan SR, Berenson GS (2009) Leukocyte telomere dynamics: longitudinal findings among young adults in the Bogalusa Heart Study. Am J Epidemiol 169:323–329CrossRefGoogle Scholar
  36. 36.
    Svenson U, Nordfjall K, Baird D, Roger L, Osterman P, Hellenius ML, Roos G (2011) Blood cell telomere length is a dynamic feature. PLoS One 6:e21485CrossRefGoogle Scholar
  37. 37.
    Gonzalo S, Jaco I, Fraga MF, Chen T, Li E, Esteller M, Blasco MA (2006) DNA methyltransferases control telomere length and telomere recombination in mammalian cells. Nat Cell Biol 8:416–424CrossRefGoogle Scholar
  38. 38.
    Bull CF, Mayrhofer G, O’Callaghan NJ, Au AY, Pickett HA, Low GKM, Zeegers D, Hande MP, Fenech MF (2013) Folate deficiency induces dysfunctional long and short telomeres; both states are associated with hypomethylation and DNA damage in human WIL2-NS cells. Cancer Prev Res (Phila) 7:128–138CrossRefGoogle Scholar
  39. 39.
    Wong JY, De Vivo I, Lin X, Grashow R, Cavallari J, Christiani DC (2014) The association between global DNA methylation and telomere length in a longitudinal study of boilermarkers. Genet Epidemiol 38:254–265CrossRefGoogle Scholar
  40. 40.
    Li H, Hilmarsen HT, Hossain MB, Björk J, Hansteen IL, Albin M, Furu Skjelbred C, Broberg K (2013) Telomere length and LINE-1 methylation is associated with chromosomal aberrations in peripheral blood. Genes Chromosomes Cancer 52:1–10CrossRefGoogle Scholar
  41. 41.
    Morrish TA, Garcia-Perez JL, Stamato TD, Taccioli GE, Sekiguchi J, Moran JV (2007) Endonuclease-independent LINE-1 retrotransposition at mammalian telomeres. Nature 446:208–212CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Irene Pusceddu
    • 1
  • Markus Herrmann
    • 2
  • Susanne H. Kirsch
    • 1
  • Christian Werner
    • 3
  • Ulrich Hübner
    • 1
  • Marion Bodis
    • 1
  • Ulrich Laufs
    • 3
  • Stefan Wagenpfeil
    • 4
  • Jürgen Geisel
    • 1
  • Wolfgang Herrmann
    • 1
  1. 1.Department of Clinical Chemistry and Laboratory MedicineSaarland University HospitalHomburg/SaarGermany
  2. 2.Department of Clinical PathologyDistrict Hospital BolzanoBolzanoItaly
  3. 3.Department of CardiologySaarland University HospitalHomburg/SaarGermany
  4. 4.Department of Biometry and EpidemiologySaarland University HospitalHomburg/SaarGermany

Personalised recommendations