ANALYSIS OF THE ASSOCIATION OF RS4977574-POLYMORPHIC VARIANTS OF THE ANRIL GENE WITH THE DEVELOPMENT OF ACUTE CORONARY SYNDROME IN INDIVIDUALS WITH DIFFERENT BODY MASS INDEX IN THE UKRAINIAN POPULATION.

The objective was to analyze the association of rs4977574-polymorphic variants of the ANRIL gene with the development of acute coronary syndrome in individuals with different body mass index. Materials and methods. The venous blood of 429 people (234 patients with acute coronary syndrome and 195 people in the control group) was used for the study. Genotyping of patients by rs4977574-polymorphic variants of the ANRIL gene was performed by real-time polymerase chain reaction (Real-time PCR) in the presence of TaqMan assay C_31720978_30. Statistical analysis of the results of the study was performed using SPSS software (version 17.0). Results. The distribution of genotypes according to SNP rs4977574 of the ANRIL gene in the group of patients with ACS and the control group among individuals with BMI < 25 kg/m 2 does not differ. Among patients with BMI 25 kg/m 2 the genotype distribution of the rs4977574-Conclusions. Individuals with BMI > 25 kg/m 2 , which were carriers of G/G genotype had a 2 times higher risk to acquire ACS than the individuals with the dominant allele. Prospects for further research. Further research will be aimed at studying the impact of ANRIL polymorphism upon the risk of ACS development depending on other risk factors. ANRIL, rs4977574, acute coronary syndrome.–

lncRNA occupy a large proportion of the protein-noncoding genome and may be involved in various critical biological processes, such as protein transport, transcription, translation, participation in cell differentiation, etc., which implies their impact on a wide range of multifactorial human diseases [6][7][8][9][10].
Scientists are especially attracted by ANRIL (Antisense Non-coding RNA in the INK4 Locus) or CDKN2B-AS1 (Cyclin-dependent kinase inhibitor 2B antisense RNA 1) -lncRNA, which consists of 3834 nucleotides and is transcribed from the antisense chain of the gene cluster INK4b-ARF-INK4a [11,12].rs4977574-polymorphism of this gene is one of the most studied and, according to different researches, is connected to lots of multifactorial diseases [15,16].For example, a recent study by Huang et al. found an association of SNP rs4977574 with overall risk of carcinogenesis [13], and Qiao et al. found an effect on glucose metabolism and the development of type 2 diabetes [14].
Probably, since the occurrence of acute coronary syndrome (ACS) depends on both genetic factors and environmental factors, and also occupies one of the first places in prevalence, this disease is a multifactorial disease, the study of which is particularly promising today.A metaanalysis of several studies has shown that SNP rs4977574 CDKN2BAS gene has a strong association with ACS in American-Caucasian and European groups [17].Several broad genomic studies (Genome-wide association study (GWAS)) found that rs4977574-polymorphism is also associated with the development of ACS and myocardial infarction in different ethnic groups [18,19].
Given the extremely large number of ACS patients diagnosed each year and the prevalence of both bad habits and sedentary and irresponsible lifestyles, studying the relationship between genetic factors and other risk factors is incredibly important.
The aim of the study.Analyze the distribution rs4977574-polymorphic variants of the ANRIL gene in individuals with acute coronary syndrome with different body mass index.

Materials and methods
The study involved 429 patients, of whom 195 were patients with ACS and 234the control group.
All patients with ACS were treated in the cardiology departments of Sumy regional clinical hospital for war veterans and Sumy regional clinical hospital.The diagnosis of acute myocardial infarction and unstable angina was established on the basis of clinical, biochemical and ECG examination in accordance with the recommendations of the European Society of Cardiology [20].
Patients with cardiogenic shock, severe renal and hepatic insufficiency, bronchial asthma, trauma or major surgery, acute or chronic inflammation in the acute phase, malignant tumors and systemic diseases were excluded from the study.
The control group included relatively healthy individuals.The absence of cardiovascular pathology was confirmed by collection of anamnestic data, ECG recording, blood pressure measurement and study of blood biochemical parameters. The

Results and discussion
Distribution of alleles and genotyping by rs4977574 polymorphism of the ANRIL gene in the experimental and control groups and the results of their comparison are presented in Table 1.The first stage of statistical processing of the results was to check the distribution of allelic variants and alleles by rs4977574 polymorphic variant of the ANRIL gene in the control group and in patients with ACS according to Hardy-Weinberg and according to Table 1 this distribution corresponds to equilibrium as in the control group (p = 0.276), and in the group of patients with ACS (p = 0.058).There was a statistically significant difference in the ratio of polymorphic variants (P = 0.035), and at the same time, the distribution of A-and G-alleles SNP rs4977574 ANRIL gene in patients with ACS significantly different from those in the control group (p = 0.008).Note: nnumber of patients; X 2 і P HWE reflect the deviations in each group from the Hardy-Weinberg equilibrium; Pstatistically significant differences in the distribution of alleles and genotypes between comparison groups Using the logistic regression method presented in Table 2, it was found that before adjusting for non-genetic risk factors, the association of rs4977574 polymorphic variant with the development of ACS was established according to recessive (P = 0.015) and additive (P = 0.012) inheritance models.Thus, homozygotes with a recessive G/G genotype are approximately 2 times more likely to develop ACS than A-allele carriers.After adjusting for the sex and age of patients, BMI, smoking habits, diabetes and stress, the identified risk remains (Р = 0.049; Р = 0.037 accordingly).The of logistic regression did not reveal a difference in the risk chances in any model of inheritance both before and after the adjustments to the sex, smoking habit, the presence of diabetes, stress (Table 4).
lncRNA ANRIL located on the short arm of chromosome 9 (9p21.3)where it and three other protein-coding genes make up the INK4b-ARF-INK4a gene cluster, in which p14ARF, p15INK4b and p16INK4a are protein suppressors of tumors that affect cell cycle delay [21].The known biological effects of ANRIL include its association with proliferation, apoptosis and cellular adhesion pathways [22].Note: 95% CI -95% confidence interval; P observobserved value P (without adjustments for covariants); OR observobserved ratio of chances; P adjust -P values after adjusting for smoking, gender, stressful occupation, diabetes and hypertension; OR adjustratio of chances after adjustments for covariants SNP rs4977574 A/G ANRIL gene is located in the position of 103785 16 th intron.The pathophysiological path of its influence on pathological processes and disease is still unknown, but the role of rs4977574 polymorphism in the development of cardiovascular diseases (CVD) has been proven by many studies around the world.Ibdah et al. in their study established the association of this polymorphism with CVD in the Jordanian population [23].A large metaanalysis found a link between the polymorphism of rs4977574 A/G ANRIL gene in the Asian and Caucasian ethnic groups [24].The study by Huang et al. indicates that SNP rs4977574 is a risk factor for ACS among women in the Asian population [25].However, when studying the association of the Chinese population with BMI < 24 kg/m 2 and > 24 kg/m 2 and rs4977574, no significant difference in the distribution of genotypes was found [26].
It should be noted that studies that would establish a link between the rs4977574polymorphic variant of the ANRIL gene and other risk factors are currently insufficient to accurately establish the role of this polymorphism.

CONCLUSIONS/ВИСНОВКИ
1.There is a significant difference in the distribution of genotypes А/А, А/G, G/G by rs4977574 ANRIL gene polymorphism among ACS patients and control group.
2. Recessive G/G genotype homozygotes are approximately 2 times more likely to develop ACS than A-allele carriers.After adjusting for the sex and age of patients, BMI, smoking habits, diabetes and stress, the identified risk remains.
3. The distribution of genotypes according to SNP rs4977574 of the ANRIL gene in the group of patients with ACS and the control group among individuals with and BMI <25 kg/m2 does not differ.
4. Among patients with BMI>25 kg/m2, the distribution of genotypes by rs4977574 polymorphic variant of the ANRIL gene is statistically significant.
5. Individuals with a BMI>25 kg/m2 who are carriers of the G/G genotype have a double risk of developing ACS than individuals with a dominant allele.

Further
research will be aimed on studying the impact of ANRIL polymorphism upon the risk of ACS development depending on other risk factors.This work is licensed under Creative Commons Attribution 4.0 International License https://creativecommons.org/licenses/by/4.0/

How to сite/ Як цитувати статтю: Kniazkova PV, Harbuzova VYu.
Analysis of the association of rs4977574-polymorphic variants of the ANRIL gene with the development of acute coronary syndrome in individuals with different body mass index in the Ukrainian population.EUMJ.