Considering the multifactorial nature of the development of coronary artery calcificatioN (CAD), and the difficulty in making its early diagnosis, this study sought to investigate the role of the TREML4 in the development of atherosclerotic lesions by genotyping of two main variants associated with this gene (rs2803495 and rs2803496), mRNA expression and identification of its regulatory miRNAs. 329 patients with Subclinical atherosclerosis (AS) were recruited. The results with AS patients stratified according to the Duke score demonstrated an association of the Duke score with increased expression of mRNA of the TREML4 in peripheral blood leukocytes (OR. = 3.173, 95% CI = 1.29-7, 78, p = 0.012). Furthermore, genotyping revealed that the SNP genotype combination rs2803495 (AG + GG) (OR = 9,007, 95% CI = 4.1-19.8, p <0.001) and rs2803496 (CT + CC) ( OR = 11.1, 95% CI = 4.8-25.4, p = 0.011), favor the expression of TREML4 mRNA. Patients with AS and carriers of the SNP G alleles rs2803495 (AG + GG) (OR = 13.31, 95% CI = 5, 79-30.61, p <0.001) and C allele rs2803496 (CT + CC) (OR = 18.99, 95% CI = 7.60-47.44, p <0.001) are more likely to express TREML4 . Once again, the C allele was associated with higher levels of mRNA expression (OR = 3.98, 95% CI = 1.67-9.48, p = 0.002). The rs2803495 (AG + GG) and rs2803496 (CT + CC) variants were not related to the development of type 2 Diabetes Mellitus (DM2; p> 0.05). However, patients who express TREML4 and are DM2 have higher levels of expression of TREML4 when compared to the control group (OR = 2.6, 95% CI = 1.19-5.72, p <0.001). Patients with DM2 and rs2803495 (AG + GG) genotypes are more likely to express TREML4 (OR = 20.77, 95% CI = 2.29-188.41, p = 0.007). In silico analyzes showed that 8 miRNAs (hsa-miR-181a-5p, hsa-miR-200b-3p, hsa-miR-24-3p, hsa-miR-296-5p, hsa-miR-361-5p, hsa-miR- 423-5p, hsa-miR-486-3p and hsa-miR-708-5p) are expected to interact with TREML4 mRNA.The results suggest that mRNA expression of the TREML4 is associated with the presence of SNPs and increased in patients with AS with DM2 and that miRNAs may be involved in the regulation of the TREML4 being directly or indirectly involved in the pathophysiology of CVD being potential circulating biomarkers for early detection and/or individualized treatment of AS and CAD.
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