LEVELS OF CIRCULATING MICRORNA-21-5P IN CHILDREN WITH TYPE 1 DIABETES MELLITUS

Objective. To determine the levels of circulating microRNA-21-5p in various fractions of blood plasma in children with type 1 diabetes mellitus (DM1) and to study their relationship with the clinical and laboratory characteristics. Design and methods. The study included 12 patients with confirmed DM1 (mean age 12 years, boys/girls = 6/6). The control group consisted of 12 healthy children without diabetes mellitus and other metabolic disorders) (mean age 9 years, boys/girls = 5/7). Ultracentrifugation was used to separate blood plasma into the exosome fraction and the fraction without membrane vesicles. A quantitative analysis of the levels of mature microRNA-21-5p in plasma fractions was carried out by real-time polymerase chain reaction. Results. In the control group, age is not associated with either the levels of microRNA-21-5p in the exosome fraction, or with the levels of microRNA-21-5p in the supernatant fraction. In patients with DM1, the age is not associated with the level of microRNA-21-5p in the exosome fraction. In the supernatant fraction, the level of microRNA-21-5p negative correlated with the age of patients with DM1 (r = –0,75; p = 0,005), as well as with the glycated hemoglobin level (r = –0,59, p = 0,045). In children with DM1 aged 10 years and older, a 1,6-fold decrease (p = 0,026) in the levels of microRNA-21-5p from the supernatant fraction compared to the control group is observed. There were no differences in the levels of microRNA-21-5p between the sex-different cohorts of children, both in the control group and among patients with diabetes mellitus. Conclusions. In 2–16-year-old children with DM1, levels of circulating extravesicular microRNA-21-5p correlate with age and the level of glycated hemoglobin. DM1 is accompanied by a decrease of circulating extravesicular microRNA-21-5p levels in 10–16 years old children.

circulating microRNA, age, children, type 1 diabetes mellitus, endothelial dysfunction, exosomes, biomarkers

1. Grover-Paez F, Zavalza-Gomez AB. Endothelial dysfunction and cardiovascular risk factors. Diabetes Res Clin Pract. 2009;84 (1):1–10. doi:10.1016/j.diabres.2008.12.013

2. Nathan DM. Long-term complications of diabetes mellitus. N Engl J Med. 1993;328(23):1676–85. doi:10.1056/ NEJM199306103282306

3. Pomilio M, Mohn A, Verrotti A, Chiarelli F. Endothelial dysfunction in children with type 1 diabetes mellitus. J Pediatr Endocrinol Metab. 2002;15(4):343–361.

4. Wang J, Liew OW, Richards AM, Chen YT. Overview of MicroRNAs in Cardiac Hypertrophy, Fibrosis, and Apoptosis. Int J Mol Sci. 2016;17(5):749.

5. LaPierre MP, Stoffel M. MicroRNAs as stress regulators in pancreatic beta cells and diabetes. Mol Metab. 2017;6(9):1010– 1023.

6. Krol J, Loedige I, Filipowicz W. The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet. 2010;11(9):597–610. doi:10.1038/nrg2843

7. Creemers EE, Tijsen AJ, Pinto YM. Circulating microRNAs: novel biomarkers and extracellular communicators in cardiovascular disease? Circ Res. 2012; 110(3):483–495. doi:10.1161/CIRCRESAHA.111.247452

8. Reid G, Kirschner MB, van Zandwijk N. Circulating microRNAs: Association with disease and potential use as biomarkers. Crit Rev Oncol Hematol. 2011;80(2):193–208. doi:10.1016/j.critrevonc.2010.11.004

9. Hu J, Ni S, Cao Y, Zhang T, Wu T, Yin X et al. The angiogenic effect of microRNA-21 targeting TIMP3 through the regulation of MMP2 and MMP9. PLoS One. 2016;11(2): e0149537. doi:10.1371/journal.pone.0149537.

10. Кондратов К. А., Петрова Т. А., Михайловский В. Ю., Иванова А. Н., Костарева А. А., Федоров А. В. Изучение внеклеточных везикул, выделенных из плазмы крови, с помощью сканирующей электронной микроскопии низкого напряжения. Цитология. 2017;59(3)169–177. [Kondratov KA, Petrova TA, Mikhailovskii VU, Ivanova AN, Kostareva AA, Fedorov AV. Extracellular vesicles from blood plasma studied by low voltage scanning electron microscopy. Tsitologiia = Cytology. 2017; 59 (3)169–177. doi:10.1134/S1990519X17030051. In Russian]

11. Kondratov K, Kurapeev D, Popov M, Sidorova M, Minasian S, Galagudza M et al. Heparinase treatment of heparin-contaminated plasma from coronary artery bypass grafting patients enables reliable quantification of microRNAs. Biomolecular detection and quantification 2016;8:9–14. doi:10.1016/j.bdq.2016.03.001

12. Meder B, Backes C, Haas J, Leidinger P, Stahler C, Großmann T et al. Influence of the confounding factors age and sex on microRNA profiles from peripheral blood. Clin Chem. 2014;60 (9):1200–1208. doi:10.1373/clinchem.2014.224238

13. Noren Hooten N, Fitzpatrick M, Wood WH 3rd, De S, Ejiogu N, Zhang Y et al. Age-related changes in microRNA levels in serum. Aging (Albany NY). 2013;5(10):725–740. doi:10.18632/aging.100603

14. Ameling S, Kacprowski T, Chilukoti RK, Malsch C, Liebscher V, Suhre K et al. Associations of circulating plasma microRNAs with age, body mass index and sex in a populationbased study. BMC Med Genomics. 2015;8:61. doi:10.1186/s12920– 015–0136–7

15. Olivieri F, Spazzafumo L, Santini G, Lazzarini R, Albertini MC, Rippo MR et al. Age-related differences in the expression of circulating microRNAs: miR-21 as a new circulating marker of inflammaging. Mech Ageing Dev. 2012;133(11–12):675–685. doi:10.1016/j.mad.2012.09.004

16. Olivieri F, Bonafè M, Spazzafumo L, Gobbi M, Prattichizzo F, Recchioni R et al. Age- and glycemia-related miR-126–3p levels in plasma and endothelial cells. Aging (Albany NY). 2014;6(9):771– 787. doi:10.18632/aging.100693

17. Zampetaki A, Kiechl S, Drozdov I, Willeit P, Mayr U, Prokopi M et al. Plasma microRNA profiling reveals loss of endothelial miR-126 and other microRNAs in type 2 diabetes. Circ Res. 2010;107(6):810–817. doi:10.1161/CIRCRESAHA.110.226357

18. Ambayya A, Su AT, Osman NH, Nik-Samsudin NR, Khalid K, Chang KM et al. Haematological reference intervals in a multiethnic population. PLoS One. 2014;9(3): e91968. doi:10.1371/journal.pone.0091968