Polymorphic variants of Caspase genes (8 & 3) in the risk prediction of Coronary Artery Disease.

Apoptosis has been involved in a number of pathological conditions including coronary artery disease (CAD). Caspases (CASP) are important regulators and executioners in both extrinsic and intrinsic apoptotic pathways. The aim of the present study is to examine the role of Caspase 8 and 3 polymorphisms in the pathogenesis of CAD. CAD patients (n=300) and healthy controls (n=300) were genotyped for polymorphisms in CASP8 (-652 6N del/ins, IVS12-19G>A), CASP3 (rs4647601;G>T) by PCR-RFLP. Splicing defects were determined by HSF. Gene interactions, Linkage disequilibrium and haplotype analysis were carried out by MDR analysis and Haploview software respectively. Molecular analysis revealed that insertion genotype (II) of CASP8 -652 6N del/ins and TT genotype of CASP3 rs4647601;G>T polymorphism conferred risk for the development of CAD. HSF analysis showed that intronic cryptic donor site for CASP8 -652 6N del/ins and a new ESE site for CASP3 rs4647601;G>T polymorphisms. SNP combinations of Caspase 8 and 3 were in perfect LD (D'=1) in controls. D-A, I-G haplotypes of Caspase 8 polymorphisms (-652 6N del/ins & IVS12-19G>A) were found to be significantly predominant in the disease group. The present study suggests that CASP8 & 3 polymorphic variants might be used as markers for susceptibility to CAD.

MMP 1 circulating levels and promoter polymorphism in risk prediction of coronary artery disease in asymptomatic first degree relatives.

Coronary artery disease (CAD) remains to be the prominent health problem in India, and its incidence is growing in developing countries as well. Matrix metalloproteinase 1 (MMP 1) is highly expressed in disruption-prone shoulder regions of the fibrous plaques. The present study aims to investigate association of MMP 1 gene polymorphisms (-1607 1G/2G) and serum circulating levels with CAD. The study includes 300 CAD patients, 100 FDRS, and 300 controls. ELISA and PCR-RFLP were performed to determine MMP 1 serum levels and genotypes respectively. MMP1 levels were high in CAD patients, followed by FDRS compared to controls (2.15±1.2ng/ml; 1.46±1.04ng/ml and 0.96±0.53ng/ml) respectively. ROC analysis showed the AUC at 95% CI of serum MMP-1 to be 0.83 and 0.73-0.94, respectively. The optimal cut-off point (sensitivity; specificity) of serum MMP 1 was >1.5ng/ml (0.74; 0.90). The 2G/2G genotype was associated with high MMP 1 circulating levels in CAD patients, and a similar trend was observed in FDRS and controls. The pre-mRNA secondary structure of the 2G allele is much more stable than 1G allele. Our results suggest MMP 1 serum levels and polymorphism as potential independent prognostic markers for future cardiovascular events. These may also help to stratify CAD patients and to identify susceptibility for CAD in asymptomatic healthy FDRS.

Genetic polymorphisms of eNOS (-786T/C, Intron 4b/4a & 894G/T) and its association with asymptomatic first degree relatives of coronary heart disease patients.

BACKGROUND: The eNOS gene has been extensively screened for important variations in the promoter (-786T/C), Intron 4 (4b/4a) and exon 7 (894G/T) regions in multiple diseases. Prevalence of Coronary Heart Disease (CHD) is much higher in young Indians when compared to other ethnic groups and first-degree relatives (FDRS) of CHD patients are at risk correlating with earlier age-of-onset and poor prognosis. OBJECTIVE: The objective of this study is to understand the genetic mechanism of eNOS -786T/C, Intron 4b/4a and 894G/T polymorphisms/haplotypes in the development of CHD and to identify individuals having coronary risk factors among the FDRS of CHD patients by comparing them with the general population. MATERIALS & METHODS: The study population consisted of 300 CHD patients with angiographically documented, 100 FDRS and 300 age and sex matched healthy controls. Genotyping of eNOS -786T/C, Intron 4b/4a was carried out by Allele specific-Polymerase Chain Reaction (As-PCR) and 894G/T by PCR-restriction fragment length polymorphism (PCR-RFLP) assays. RESULTS: The frequency of risk factors for CHD includes age, blood pressure, obesity, diabetes mellitus, smoking, non-vegetarian diet, family history are higher in CHD patients and FDRS compared to controls (p < 0.01). The genotype distribution, allele frequencies and haplotypes of the eNOS -786T/C, 4b/4a and 894G/T were significantly different between CHD patients, FDRS and controls. eNOS gene haplotypes GCB, TCB (G-allele of 894G/T, C-allele -786T/C, B-allele of Intron 4b/4a respectively) were associated with high nitrite/nitrate levels compared to GTB in both FDRS and CHD patients (p < 0.01). CONCLUSION: The study shows the importance of eNOS polymorphism in the pathogenesis of CHD and predicting the risk of future coronary events in asymptomatic healthy FDRS and suggests the use of new therapeutics and initiating the measures for prevention or delaying the progression of the disease by life style modification or treatment modalities in high-risk individuals.

A Therapeutic Effects of Atorvastatin on Genetic Damage in Coronary Artery Disease.

INTRODUCTION: Coronary Artery Disease (CAD) a multifactorial chronic heart disease and the most frequent cause of death and disabling symptoms worldwide, occurs due to the formation of atheromatous lipid rich plaques in the arteries. Statins, which inhibit cholesterol biosynthesis, have both pleiotropic and Low-Density Lipoprotein (LDL)-lowering properties. Atorvastatin is one of the choices for the primary and secondary prevention of cardiovascular disease and management of hypercholesterolaemia. However, there is a paucity of data regarding the changes in the DNA damage in patients with coronary atherosclerosis after statin use. AIM: The aim of the present study was to evaluate atorvastatin treatment efficacy on lipid profiles and DNA damage in CAD patients. MATERIALS AND METHODS: The current observational study was conducted on 180 CAD patients between November 2011 to December 2013 at Durgabai Deshmukh Hospital and Research Centre, Vidya Nagar, Hyderabad, India. Atorvastatin administered and blood samples were collected at index hospitalization and after 6 months statin therapy and lipid profiles and DNA damage was compared with 200 healthy control. RESULTS: Lipid profiles and DNA damage were found to be significantly high (p < 0.01) in CAD patients before atorvastatin therapy compared to after 6 months statin therapy and healthy controls. CONCLUSION: The study suggests that atorvastatin might help in regression of lipid profile as well as DNA damage of CAD patients.

Influence of gelatinase B polymorphic variants and its serum levels in atherosclerosis.

AIM: Atherosclerosis, the underlying pathology of cardiovascular disease, is a common, multifactorial disorder with both genetic and environmental components as risk factors. Gelatinase B, also known as MMP-9, is one of the matrix metalloproteinases that is highly expressed in the disruption-prone regions of atherosclerotic plaques. It has been hypothesized that a genetic variation affecting the expression or activity of MMP-9 influences the susceptibility and progression of atherosclerosis. The present study aims to ascertain the polymorphic variants of the MMP-9 gene promoter and its serum levels, which contribute to interindividual differences in susceptibility to atherosclerosis. The study population consisted of 200 individuals who include 100 cases with angiographically recorded coronary artery disease (CAD) and 100 age- and sex-matched healthy controls. Serum levels of MMP-9 were determined in these subjects using an enzyme-linked immunosorbent assay, and polymorphic genotypes of MMP-9 were determined by the polymerase chain reaction-restriction fragment length polymorphism assay. RESULTS: The MMP-9 levels among subjects with the TT genotype for controls (12.21±2.39) and CAD (22.86±2.45) were significantly higher than that of CC genotype (controls 10.37±1.42 and CAD 16.44±7.99), and the values were intermediate for the CT genotype (control 11.21±2.01 and CAD 18.80±3.17) and found to be significant at p<0.01. Genotypic analysis of -1562C/T polymorphism among patients and controls showed higher T allele frequencies in the patient group (0.36) than in the controls (0.29). CONCLUSIONS: It has been observed that increased MMP-9 expression in T allele carriers may contribute to the severity of coronary atherosclerosis. These findings not only are relevant to the understanding of the pathogenesis of atherosclerosis but also may provide a novel target for future development of predictive, preventive, and therapeutic measures.