Strong Association of Angiotensin Converting Enzyme-2 Gene Insertion/Deletion Polymorphism with Susceptibility to SARS-CoV-2, Hypertension, Coronary Artery Disease and COVID-19 Disease Mortality

Abstract

Background: The ongoing outbreak of SARS-CoV-2 represents a significant challenge to international health. Several reports have highlighted the importance of ACE2 on the pathogenesis of COVID-19. The spike protein of SARS-CoV-2 efficiently binds to the angiotensin-converting enzyme 2 (ACE2) receptors and facilitates virus entry into the host cell. In the present study, we hypothesize that a functional insertion/deletion polymorphism-rs4646994 I/D and rs4240157 T > C in the ACE gene could be associated with SARS-CoV-2 infection and mortality. Methodology: This study included 117 consecutive COVID-19 patients and 150 age matched healthy controls (ACE2-rs4646994 I/D) and 100 age matched healthy controls with ACE2 rs4240157 T > C. We used Mutation specific PCR (MSP) for ACE2-rs4646994 I/D genotyping and amplification refractory mutation system (ARMS-PCR) for ACE2 rs4240157 T > C genotyping. Results: Results indicated that there were significant differences in the genotype distributions of ACE2-rs4646994 I/D polymorphisms (p < 0.030) and ACE2 rs4240157 T > C between COVID-19 patients and controls (p-values < 0.05). Higher frequency of DD genotype (48.71%) and D allele (0.67) was reported in COVID-19 patients than controls. Our results showed that the ACE2-DD genotype was strongly associated with increased COVID-19 severity (OR 2.37 (95%) CI = (1.19–4.70), RR = 1.39 (1.09–1.77), p < 0.013) and also a strong association was seen with ACE2-ID genotype with COVID-19 severity (OR 2.20 (95%) CI = (1.08–4.46), p < 0.020) in the codominant model. In allelic comparison, the D allele was strongly associated with COVID-19 severity (OR 1.58 (95% CI) (1.11–2.27), RR 1.21 (1.05–1.41) p < 0.010). A significant correlation of ACE2-I/D genotypes was reported with Age (p < 0.035), T2D (p < 0.0013), hypertension (p < 0.0031) and coronary artery disease (p < 0.0001). Our results indicated ACE2-DD genotype was strongly associated with increased COVID-19 mortality (OR 8.25 (95%) CI = (2.40 to 28.34), p < 0.008) and also ACE2-DD + DI genotype was strongly associated with increased COVID-19 mortality with OR 4.74 (95%) CI = (1.5214 to 14.7915), p < 0.007. A significant correlation was reported between COVID-19 patients and age matched controls (p < 0.0007). Higher frequency of heterozygosity TC (40%) followed by ACE2-CC genotype (24.78%) was reported among COVID-19 patients. Using multivariate analysis, ACE2–CT genotype was strong associated with SARS-CoV-2 severity with an OR 2.18 (95% CI) (1.92–3.99), p < 0.010 and also ACE2–CC genotype was linked with COVID-19 severity with an OR 2.66 (95% CI) (1.53–4.62), p < 0.005. A significant correlation of ACE2-T > C genotypes was reported with gender (p < 0.04), T2D (p < 0.035). ACE2-CC genotype was strongly associated with increased COVID-19 mortality OR 3.66 (95%) CI = (1.34 to 9.97), p < 0.011 and also ACE2-C allele was associated with COVID-19 mortality OR 2, 01 (1.1761–3.45), p < 0.010. Conclusions: It is concluded that ACE-DD genotype and D allele was strongly associated with increased COVID-19 patient severity. In addition, ACE I/D polymorphism were strongly associated with advanced age, diabetes and ischemic heart disease in COVID-19 patients whereas ACE-II genotype was a protective factor against the development of severe COVID-19. ACE2-DD genotype was strongly associated with increased COVID-19 mortality. Additionally, ACE2–CC and CT genotypes were strongly associated with COVID-19 severity. Therefore, our study might be useful for identifying the susceptible population groups for targeted interventions and for making relevant public health policy decisions.

Clinical Utility of Amplification Refractory Mutation System-Based PCR and Mutation-Specific PCR for Precise and Rapid Genotyping of Angiotensin-Converting Enzyme 1 (ACE1-rs4646996 D>I) and Angiotensin-Converting Enzyme 2 (ACE2-rs4240157T>C) Gene Variations in Coronary Artery Disease and Their Strong Association with Its Disease Susceptibility and Progression

“…However, the higher incidence of heterozygosity was reported among coronary artery disease patients who were younger than 50 years old with MI, which suggests that the contribution of genetics as a risk factor for MI occurrence became stronger among younger individuals—having none of the traditional risk factors, such as age. Similar results were reported by MM et al [ 46 ], which showed a significant correlation of ACE2-I/D genotypes with respect to age ( p < 0.035). On the other hand, there was a study done by Maria-Cristina Vladeanu et al [ 47 ] that reported no statistically significant differences.…”

“…Studies noted that smoking and the D allele increase the superoxide anion formation and degradation of the nitric oxide that cause endothelial dysfunction favoring the occurrence of coronary disease [ 45 ]. Significant differences in the genotype distribution were detected only in those children with 2 or more affected grandparents This research is suggestive that a family history of CAD and ACE I/D genotype are bidirectionally linked, whereas recent data has shown that the DD genotype and a family history of CAD are independent risk factors for CAD, and, further, that the DD genotype may increase the risk to those who also share a family history of CAD [ 46 ].…”

Association Between the LZTFL1 rs11385942 Polymorphism and COVID-19 Severity in Colombian Population

“…Such discrepancies might be explained by population-specific differences, the additive role of other genes interacting with risk alleles or other mechanisms not assessed such as epigenetic modifiers ( 98 – 100 ). Concerning ACE rs4646994 the Del allele has been associated with increased ACE expression, higher enzyme activity and elevated production of angiotensin II ( 101 ). Despite ACE Del/Del genotype and Del allele have been associated with increased COVID-19 patient severity ( 101 – 103 ), our results failed to replicate these findings in the Colombian population.…”

“…Concerning ACE rs4646994 the Del allele has been associated with increased ACE expression, higher enzyme activity and elevated production of angiotensin II ( 101 ). Despite ACE Del/Del genotype and Del allele have been associated with increased COVID-19 patient severity ( 101 – 103 ), our results failed to replicate these findings in the Colombian population. In agreement with our results, other studies have reported no association between ACE rs4646994 and COVID-19 severity ( 43 , 96 ).…”

Biochemical Characterization and Molecular Determination of Estrogen Receptor-α (ESR1 PvuII-rs2234693 T>C) and MiRNA-146a (rs2910164 C>G) Polymorphic Gene Variations and Their Association with the Risk of Polycystic Ovary Syndrome

“…Adjusted odds ratios (OR) and 95% confidence intervals (95% CI) associated with the risk of PCOS, were analyzed by logistic regression after controlling for a number of covariates and comparison with the reference group (healthy controls). Multivariate analysis was used to study the link between miR- 146a rs2910164 C>G and ESR1 PvuII-rs2234693 T>C genotyping and the susceptibility to disease in terms of odds ratios (ORs), risk ratios (RRs) and risk differences (RDs) with 95% confidence intervals (CIs) [ 38 , 39 ].…”