Polymorphism of the APOA5 gene in patients with primary hyperlipidemia

Aim. To study APOA5 genetic polymorphism in Caucasoid patients with familial hypercholesterolemia.

Methods. Sample of patient with familial hypercholesterolemia (43 unrelated Caucasoid persons) was formed using Dutch Lipid Clinic Network Criteria. Targeted sequencing of genome DNA was performed by NimbleGen SeqCap EZ Choice kit on pyrosequencer Roche Junior GS (Roche, Switzerland).

Results. In patients with familial hypercholesterolemia, 8 substitutions were identified in the APOA5 gene: rs2075291, rs3135506, rs2072560, rs2266788, rs3135507, rs34089864, rs619054, and rs651821, that are known to be associated with dyslipidemia. One novel substitution Ala169Asp was found. It is responsible for changing the charge of a domain for lipid droplets binding in the APOA5 protein. There were no differences in the frequencies of the ApoA5*2 intragenic haplotype, which has been recently reported to be associated with an increased triglyceride levels in patients with familial hypercholesterolemia and the population.

Conclusion. Genetic variants ofAPOA5, common in patients with familial hypercholesterolemia, may be involved in the formation of the pathological phenotype of dyslipidemia. However, a more accurate assessment oftheir contribution is required to differentiate patients with familial hypercholesterolemia according to their triglycerides level.

1. Do R., Stitziel N.O., Won H.H., J0rgensen A.B., Duga S., Merlini A.P., et al. Exome sequencing identifies rare LDLR and APOA5 alleles conferring risk for myocardial infarction. Nature. 2015; 518:102-106. doi: 10.1038/nature13917

2. Ramasamy I. Update on the laboratory investigation of dyslipidemias. Clin Chim Acta. 2018; 479:103-125. doi: 10.1016/j.cca.2018.01.015.

3. Su X., Kong Y, Peng D.Q. New insights into apolipoprotein A5 in controlling lipoprotein metabolism in obesity and the metabolic syndrome patients. Lipids Health Dis. 2018; 17(1):174. doi: 10.1186/s12944-018-0833-2.

4. Shakhtshneider E.V., Ivanoshchuk D.E., Makarenkova K.V., Orlov P.S., Timoshchenko O.V., Bazhan S.S., Nikitin Yu.P.,Voevoda M.I Cascade genetic screening in diagnostics of heterozygous familial hypercholesterolemia: clinical case. Russ J Cardiol. 2017; 6 (146): 178-179. (In Russian). https://doi.org/10.15829/1560-4071-2017-6-178-179.

5. Mikhailova S., Ivanoshchuk D., Timoshchenko O., Shakhtshneider E. Genes Potentially Associated with Familial Hypercholesterolemia. Biomolecules. 2019; 9(12). pii: E807. doi: 10.3390/biom9120807.

6. Pecherina T.B., Khorlampenko A.A., Gorbunova E.V, Strokolskaya I.L., Kashtalap VV. Clinical case o f a patient with heterozygousfamily hypercholesterolemia. Complex Issues of Cardiovascular Diseases. 2017;6(4): 155-168. (In Russian). DOI:10.17802/2306-1278-2017-6-4-lSS-168.

7. Pennacchio L.A., Olivier M., Hubacek J.A., Cohen J.C., Cox D.R., Fruchart J.C., et al. An apolipoprotein influencing triglycerides in humans and mice revealed by comparative sequencing. Science. 2001; 294(5540):169-173. DOI: 10.1126/science.1064852

8. Shu X., Nelbach L., Ryan R.O., Forte T.M. Apolipoprotein A-V associates with intrahepatic lipid droplets and influences triglyceride accumulation. Biochim Biophys Acta. 2010; 1801:605-608. doi: 10.1016/j.bbalip.2010.02.004.

9. Merkel M., Heeren J. Give me A5 for lipoprotein hydrolysis! J Clin Invest. 2005; 115(10):2694-2696. DOI: 10.1172/JCI26712.

10. Guardiola M., Alvaro A., Vallve J.C., Rosales R., Sola R., Girona J., et al. APOA5 gene expression in the human intestinal tissue and its response to in vitro exposure to fatty acid and fibrate. Nutr Metab Cardiovasc Dis. 2012; 22:756762. doi: 10.1016/j.numecd.2010.12.003.

11. Guardiola M., Ribalta J. Update on APOA5 Genetics: Toward a Better Understanding of Its Physiological Impact. Curr Atheroscler Rep. 2017; 19(7):30. doi: 10.1007/s11883-017-0665-y.

12. Hubacek J.A. Apolipoprotein A5 fifteen years anniversary: Lessons from genetic epidemiology. Gene. 2016; 592(1):193-199. doi: 10.1016/j.gene.2016.07.070.

13. Pennacchio LA, Olivier M, Hubacek JA, Krauss RM, Rubin EM, Cohen JC. Two independent apolipoprotein A5 haplotypes influence human plasma triglyceride levels. Hum Mol Genet. 2002; 11(24):3031-3038. DOI: 10.1093/hmg/11.24.3031.

14. Melegh B.I., Duga B., Sumegi K., Kisfali P., Maasz A., Komlósi K., et al. Mutations of the apolipoprotein A5 gene with inherited hypertriglyceridaemia: Review of the current literature. Curr Med Chem. 2012; 19(36):6163-6170. DOI: 10.2174/092986712804485719.

15. Caussy C., Charriere S., Maręais C., Di Filippo M., Sassolas A., Delay M., et al. An APOA5 3' UTR variant associated with plasma triglycerides triggers APOA5 downregulation by creating a functional miR-485-5p binding site.Am J Hum Genet. 2014; 94(1):129-134. doi: 10.1016/j.ajhg.2013.12.001.

16. Defesche J.C., Lansberg P.J., Umans-Eckenhausen M.A., Kastelein J.J. Advanced method for the identification of patients with inherited hypercholesterolemia. Semin Vasc Med. 2004; 4:59-65.

17. Sambrook J., Russell D.W. Purification of nucleic acids by extraction with phenol:chloroform. CSH Protoc. 2006; 2006(1). pii: pdb.prot4455. doi: 10.1101/pdb.prot4455.

18. Qian X., Li Y, Liu X., Li L., Yang K., Liu R., et al. The "T" allele of apolipoprotein A5 rs2075291 is significantly associated with higher total cholesterol and triglyceride and lower high-density lipoprotein cholesterol levels in Asians: a meta-analysis. Nutr Res. 2018; 56:11-22. doi: 10.1016/j.nutres.2018.03.018.

19. You Y, Wu Y-H., Zhang Y, Zhang L., Song Y, Bai W., et al. Effects of polymorphisms in APOA5 on the plasma levels of triglycerides and risk of coronary heart disease in Jilin, northeast China: a case-control study. BMJ Open. 2018; 8(6): e020016. doi: 10.1136/bmjopen-2017-020016.

20. Vrablik M., Hubacek J.A., Dlouha D., Satny M., Adamkova V., Ceska R. Strong association between APOA5 gene polymorphisms and hypertriglyceridaemic tpisodes. Folia Biol (Praha). 2019; 65(4):188-194.

21. Jasim A.A., Al-Bustan S.A., Al-Kandari W, Al-Serri A., AlAskar H. Sequence analysis of APOA5 among the Kuwaiti population identifies association of rs2072560, rs2266788, and rs662799 with TG and VLDL levels. Front Genet. 2018; 9:112. doi: 10.3389/fgene.2018.00112.

22. Fahrioglu U., Ergoren M.ę. The association between APOA5 gene polymorphisms and plasma lipids in the Turkish Cypriot population: a possible biomarker for preventing cardiovascular diseases. Biochem Genet. 2018; 56(3):176-187. doi: 10.1007/s10528-017-9836-3.

23. Salehi S., Emadi-Baygi M., Rezaei M., Kelishadi R., Nikpour P. Lack of evidence of the role of APOA5 3UTR polymorphisms in Iranian children and adolescents with metabolic syndrome. Diabetes Metab J. 2018;42(1):74-81. doi: 10.4093/dmj.2018.42.1.74.

24. Hubacek J.A., Skodova Z., Adamkova V, Lanska V, Pitha J. APOA5 variant Ser19Trp influences a decrease of the total cholesterol in a male 8 year cohort. Clin Biochem. 2006;39(2):133-136. DOI: 10.1016/j.clinbiochem.2005.11.012

25. Williams P.T. Gene-environment interactions due to quantile-specific heritability of triglyceride and VLDL concentrations. Sci Rep. 2020;10(1):4486. doi: 10.1038/s41598-020-60965-9.