Plasma Levels of microRNA-145 Are Associated with Severity of Coronary Artery Disease

Abstract

Background and Objective

MicroRNAs (miRNAs) have been shown to be associated with various physiological and pathological conditions, including inflammation and cardiovascular disease, but little is known about their relationship with the presence of coronary artery disease (CAD) and disease severity.

Methods

A total of 195 consecutive subjects who underwent coronary angiography for chest pain evaluation were enrolled in this study. In CAD patients severity of coronary lesions was assessed by the number of diseased vessels and the Synergy between PCI with Taxus and Cardiac surgery score (SYNTAX score). Plasma levels of miRNA-145 were quantified by real-time quantitative polymerase chain reaction test, and logarithmic transformation of miRNA-145 levels (Ln_miRNA-145) was used for analyses due to its skewed distribution.

Results

Of the 195 total subjects 167 patients were diagnosed as having CAD. Ln_miRNA-145 was significantly lower in CAD patients compared with the non-CAD group (-6.11±0.92 vs. -5.06±1.25; p <0.001). In multivariable linear regression analyses CAD was significantly associated with lower Ln_miRNA-145 (Estimate, -0.50; standard error (SE), 0.11; p <0.0001). Furthermore, among CAD patients, three-vessel disease, higher SYNTAX scores and STEMI were significantly associated with lower Ln_miRNA-145 ([Estimate, -0.40; SE, 0.07; p <0.0001]; [Estimate, -0.02, SE, 0.10; p = 0.005] and [Estimate, -0.35, SE, 0.10; p <0.001] respectively).

Conclusions

Lower plasma levels of miRNA-145 were significantly associated with the presence as well as severity of CAD. As a potential biomarker for CAD, plasma miRNA-145 may be useful in predicting CAD and its severity in patients presenting with chest pain.

About the Authors:

Hai Gao

Contributed equally to this work with: Hai Gao, Raviteja Reddy Guddeti

Affiliations Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing, China, Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing, China

Raviteja Reddy Guddeti

Contributed equally to this work with: Hai Gao, Raviteja Reddy Guddeti

Affiliation: Division of Cardiovascular Diseases, Mayo College of Medicine, Rochester, Minnesota, United States of America

Yasushi Matsuzawa

Affiliation: Division of Cardiovascular Diseases, Mayo College of Medicine, Rochester, Minnesota, United States of America

Li-Ping Liu

Affiliation: Department of Nephrology, First Hospital of Tsinghua University, Beijing, China

Li-Xiao Su

Affiliation: Department of Biostatistics, School of Public Health, Rutgers, The State University of New Jersey, Newark, United States of America

Duo Guo

Affiliation: Department of Histology and Embryology, School of Basic Medical Sciences, Capital Medical University, Beijing, China

Shao-Ping Nie

Affiliations Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing, China, Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing, China

Jie Du

Affiliations Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing, China, Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing, China

Ming Zhang

* E-mail: [email protected]

Affiliations Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing, China, Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing, China

Introduction

Despite recent progress in the diagnosis and treatment of cardiovascular diseases, coronary artery disease (CAD) and its clinical residues still represent a major cause of morbidity and mortality worldwide [1–3]. Atherosclerosis, which is the major cause of CAD, is now widely considered as a chronic inflammatory state [4]. Vascular smooth muscle cells (VSMCs) play a critical role in the pathogenesis of a variety of cardiovascular diseases such as atherosclerosis, aneurysms and re-stenosis after angioplasty or bypass [5,6]. Conversion of VSMC phenotype from differentiated to dedifferentiated is known to be associated with marked increase in VSMC expression of inflammatory cytokines which facilitate monocyte and macrophage adhesion promoting plaque formation [7]. Recent studies have demonstrated that micro ribonucleic acids (miRNAs), which are small, single-stranded, non-coding RNA molecules known to be essential as post-transcriptional modulators of gene expression and coordinators of several regulatory pathways involved in cell differentiation and proliferation, are associated with various physiological and pathological conditions, including inflammation and cardiovascular disease [8–11]. The proposed mechanisms by which miRNAs regulate cellular differentiation and proliferation include either translational repression or RNA degradation [8]. MiRNAs are predicted to control and regulate approximately 60% of all coding genes. Plasma miRNAs have been considered as promising novel biomarkers for the diagnosis and prognosis of cardiovascular diseases, especially CAD [12,13].

Several lines of evidence suggest that miRNA-145 is involved in the regulation of VSMC proliferation, differentiation and phenotype [5]. MiRNA-145 is also one of the most abundant miRNA in normal, healthy arterial walls, and VSMCs and has been shown to be the major determinant of VSMC phenotype in vivo. Experimental animal models have reported that expression of miRNA-145 in arteries is attenuated in vascular injury, atherosclerosis and aneurysms [14,15]. Although expression of different circulating miRNAs in CAD patients has been reported [16], the role of miRNA-145 in atherosclerosis has not been studied extensively among different presentations of CAD.

In the present study, we aimed to determine the association between plasma miRNA-145 levels and the presence and severity of CAD.

Methods

Study subjects

Between 5/28/2012 and 12/17/2013 a total of 195 consecutive patients who underwent diagnostic coronary angiography for chest pain evaluation were recruited from inpatients admitted to Beijing Anzhen Hospital of Capital Medical University. Coronary angiography was performed via transfemoral or transradial approach using techniques, equipment and practices standard to Beijing Anzhen Hospital’s cardiac catheterization laboratory. Diagnosis of CAD was confirmed by coronary angiography and was defined as angiographic evidence of more than 50% luminal narrowing in at least one segment of a main epicardial coronary artery [17]. CAD patients were again categorized into ST-elevation myocardial infarction (STEMI), unstable angina/non-ST-elevation MI (UA/NSTEMI) and stable angina based on standard ACC/AHA definitions. STEMI and UA/NSTEMI patients constituted ACS group. Subjects with no angiographic evidence of CAD were grouped under non-CAD.

Patients with histories of significant concomitant diseases including hepatic failure, renal failure, hepatitis, cardiomyopathy, congenital heart disease, bleeding disorders, previous thoracic irradiation therapy, and malignant diseases were excluded from the study.

Ethics statement

The study protocol was approved by the Beijing Anzhen Hospital Ethics Committee of Capital Medical University, and a written informed consent was obtained from all the participants.

Plasma collection and storage

Blood samples were collected in EDTA-containing tubes (BD, Franklin Lakes, NJ, USA) and plasma was isolated within 1 hour by centrifugation at 1900×g for 10 minutes at 4°C to remove blood cells, and then at 16,000×g for 10 minutes at 4°C to remove additional cellular nucleic acids attached to cell debris, plasma samples were transferred to RNase/Dnase-free tubes and stored at -80°C. Studies have shown that heparin administration as part of ACS treatment is associated with interference in plasma miRNA measurement [18]. Therefore in our study blood samples were collected before heparin boluses were administered, especially in patients presenting with unstable CAD features. In patients who presented with stable CAD samples were collected by the next day.

RNA isolation and preparation

Total RNA in plasma was isolated by the use of miRNeasy Plasma Kit (QIAGEN, Hilden, Germany) following the manufacturer's instructions. In detail, 200 μL EDTA-containing plasma was mixed thoroughly with 1000 μL QIAzol in RNase-free tube, incubated for 5 minutes at room temperature (15~25°C). To normalize for the plasma miRNA content, we supplemented 3.5 μL (1.6 × 108 copies/μL) Caenorhabditis elegans miR-39 (cel-miR-39) to the samples (after addition of QIAzol) [19]. After mixing thoroughly, 200 μL of chloroform was added into the mix. Following vigorous vortex mix for 15 seconds, the tube containing the lysate was placed at room temperature for 2~3 minutes and then centrifuged for 15 minutes at 12000×g at 4°C. Later, the aqueous phase containing the RNA was carefully removed.

Quantification of microRNA-145

Total RNA was poly-adenylated and reverse transcribed using the miScript Reverse Transcription kit (QIAGEN) as per the manufacturer's instructions with 100~200 ng of total RNA per reaction. The obtained cDNA (2.5 μL of a 1/100 dilution) was then PCR-amplified using the miScript SYBR Green PCR kit (QIAGEN) in 25 μL reaction mixtures consisting of 12.5 μL Quantitect SYBR Green PCR master mix and 2.5ul miR-145 primer and the miScript universal primer. The reactions were incubated at 95°C for 15 minutes, followed by 45 cycles of 94°C for 15 seconds, 55°C for 30 seconds, and 70°C for 30 seconds. The relative expression level for miR-145 was computed using the comparative Ct method. MiRNA expression was normalized to cel-miR-39 and expressed as 2− (CT [microRNA] −CT [cel-miR-39]) [19]

Assessment of CAD severity

Depending on the extent of epicardial coronary artery involvement CAD patients were divided into single-, double-, and triple-vessel disease subgroups in accordance with the Coronary Artery Surgery Study classification [20,21]. Severity of coronary lesions was assessed using the Synergy between PCI with Taxus and Cardiac surgery score (SYNTAX score), an angiographic score to determine the severity and complexity of the disease [22]. Total SYNTAX score for each patient was calculated by summing the total points assigned to each individual lesion in coronary arteries >50% stenosis and >1.5 mm diameter. Two cardiologists, who were blinded to the study patient data, assessed the angiograms and SYNTAX scores were measured accordingly.

Statistical analysis

Normality of distribution was assessed using Kolmogorov-Smirnov test. In our study the distribution of plasma miRNA-145 values among the study subjects was skewed and therefore logarithmic transformation was performed to normalize plasma miRNA-145 data (Ln_miRNA-145). Comparisons between 2 groups were performed using Fisher’s test, Student’s t test or Mann–Whitney U test. For comparison of more than 2 groups, one-way ANOVA and Tukey-Kramer HSD (honestly significant difference) tests were used as appropriate. Pearson χ2 test and Spearman ρ test were used to compare qualitative and quantitative variables as appropriate. Correlations between plasma levels of miRNAs and other variables were calculated using Pearson correlation coefficient for symmetric distributions and Spearman’s coefficient for skewed variables. Univariate and multivariable linear regression analyses were performed to determine variables that were associated with lower Ln_miRNA-145. All tests were two-sided and a p <0.05 was considered statistically significant. All statistical analyses were performed using JMP 10 software (SAS Institute, Inc., Cary, NC, USA).

Results

Basic clinical characteristics of patients

Table 1 demonstrates basic clinical characteristics in the whole population. Mean age was 59.5±11 years and males constituted 70% of the total subjects. Prevalence of traditional CAD risk factors, such as hypertension, diabetes mellitus, dyslipidemia and smoking was 70%, 37%, 37% and 45% respectively (Table 1 and S1 Table). Mean Ln_miRNA-145 in the whole population was -6.0±1.0. Of the 195 patients 167 were diagnosed as having CAD and the rest of them were grouped under non-CAD (n = 28).

[Figure omitted. See PDF.]

Table 1. Basic patient characteristics in the whole population.

https://doi.org/10.1371/journal.pone.0123477.t001

Association of plasma miRNA-145 levels and traditional risk factors for CAD

Table 2 demonstrates univariate linear regression analysis for the association of clinical variables and Ln_miRNA-145. Diabetes and smoking were significantly (p = 0.032 and 0.008 respectively) and male gender was non-significantly (p = 0.070) associated with lower Ln_miRNA-145 (Table 2). Although dyslipidemia showed a tendency for negative association with miRNA-145 levels, the p value was not statistically significant (p = 0.12).

[Figure omitted. See PDF.]

Table 2. Univariate linear regression analysis for plasma levels of Ln_miRNA-145.

https://doi.org/10.1371/journal.pone.0123477.t002

Reduced plasma miRNA-145 levels are associated with the presence of CAD

CAD patients had significantly lower plasma levels of miRNA-145 compared with non-CAD subjects (Ln_miRNA-145 -6.11±0.92 vs. -5.06±1.25; p <0.001). CAD was associated with lower Ln_miRNA-145 both in univariate linear regression analysis (Estimate, -0.52; standard error, 0.1; p <0.0001) and multivariate linear regression analysis (Estimate, -0.50; standard error, 0.11; p <0.0001) after adjustment for traditional coronary risk factors (Table 3). Interestingly, in the current study plasma miRNA-145 levels significantly correlated with higher ejection fraction (Spearman ρ = 0.18; p = 0.014) and larger left ventricular end-diastolic diameter (Spearman ρ = -0.26; p < 0.001) respectively.

[Figure omitted. See PDF.]

Table 3. Association of miRNA-145 with severity of CAD.

https://doi.org/10.1371/journal.pone.0123477.t003

Association of plasma miRNA-145 levels with severity of CAD

To further evaluate the relationship between plasma miRNA-145 levels and CAD we performed a sub-analysis in CAD population (n = 167) to determine the association between plasma levels of miRNA-145 and severity of CAD. Severity of CAD was assessed by number of diseased vessels, SYNTAX score and the type of CAD at presentation. By coronary angiography, 34 (20%) CAD patients had single-vessel disease, 60 (36%) had double-vessel disease and 73 (44%) had triple-vessel disease. The levels of miRNA-145 in patients with three-vessel disease were significantly lower than those with one or two-vessel disease (Ln_miRNA-145 -6.38±0.94 vs. -5.78±0.98; p = 0.002 and -6.38±0.94 vs. -5.98±0.79; p = 0.025, respectively) (“Fig 1”). However, our study results showed that the levels between patients with one-vessel and two-vessel disease were not significantly different (p = 0.59) (“Fig 1”). In univariate (Estimate, -0.40, standard error, 0.06; p <0.0001) and multivariable linear regression (Estimate, -0.40, standard error, 0.07; p <0.0001) analyses higher number of diseased vessels was significantly associated with lower Ln_miRNA-145 (Table 3).

[Figure omitted. See PDF.]

Fig 1. Association between plasma miRNA-145 levels and number of diseased vessels.

These bars represent averages of plasma miRNA-145 levels among patients with single-, double- and triple-vessel CAD. T-bars indicate standard deviation. Patients with three-vessel disease have significantly lower levels of miRNA-145 compared with the rest of the groups. Levels were not significantly different between patients with single- and double-vessel CAD. * p <0.05; CAD, coronary artery disease, RNA, Ribonucleic acid.

https://doi.org/10.1371/journal.pone.0123477.g001

On the basis of their SYNTAX scores CAD patients were again divided into three groups; low (SYNTAX score <23; n = 125), intermediate (SYNTAX score 23–32; n = 24) and high (SYNTAX score >32; n = 18) and the plasma miRNA-145 levels in each group were assessed. Patients with high SYNTAX score had significantly lower Ln_miRNA-145 levels compared with those with low and intermediate score (Low score group -6.05±0.91, Intermediate score group -5.88±1.00, High score group -6.76±0.66; p = 0.004 [low vs. high score groups] and p = 0.005 [intermediate vs. high score groups], respectively) (“Fig 2”). Although our study data demonstrated significant differences in miRNA-145 levels between low SYNTAX score versus high score, and intermediate score versus high score groups, no significant differences were observed between low score and intermediate score groups (p = 0.66). Lower plasma levels of miRNA-145 significantly correlated with higher SYNTAX scores (Spearman ρ = -0.30; p <0.0001), indicating reduced plasma miRNA-145 levels with increase in severity and complexity of CAD (“Fig 3”). In univariate and multivariable linear regression analyses higher SYNTAX score was significantly associated with lower Ln_miRNA-145 (Estimate, -0.02; standard error, 0.006; p = 0.003 and Estimate, -0.02; standard error, 0.007; p = 0.005, respectively) (Table 3).

[Figure omitted. See PDF.]

Fig 2. SYNTAX groups and plasma miRNA-145.

These bars represent averages of plasma miRNA-145 levels across SYNTAX groups. T-bars indicate standard deviation. It can be noted that patients with high risk SYNTAX scores had significantly lower Ln_miRNA-145 compared with low and moderate risk groups. * p <0.05; RNA, Ribonucleic acid; SYNTAX, Synergy Between PCI With Taxus and Cardiac Surgery.

https://doi.org/10.1371/journal.pone.0123477.g002

[Figure omitted. See PDF.]

Fig 3. Correlation between SYNTAX score and plasma miRNA-145.

The above scatter diagram demonstrates correlation between miRNA-145 and SYNTAX score. It can be noted that increase in SYNTAX score (increase in severity and complexity of CAD) is significantly correlated with reduced plasma miRNA-145 levels. * p <0.05; CAD, Coronary artery disease; RNA, Ribonucleic acid; SYNTAX, Synergy between PCI With Taxus and Cardiac Surgery.

https://doi.org/10.1371/journal.pone.0123477.g003

Altered expression of miRNA-145 in patients with ST-elevation myocardial infarction

Levels of plasma miRNA-145 were lower in patients with ACS, especially STEMI. “Fig 4” demonstrates the relative differences in Ln_miRNA-145 values between non-CAD (n = 28), stable angina pectoris (n = 26), UA/NSTEMI (n = 106) and STEMI (n = 35). It can be noted that STEMI patients had significantly lower Ln_miRNA-145 compared with the other groups indicating an altered expression of miRNA-145 in these patients. STEMI was associated with significantly lower Ln_miRNA-145 in multivariable linear regression analysis after adjustment for traditional CAD risk factors (Estimate, -0.35; standard error, 0.10; p <0.001 respectively) (Table 3). However, in our study miRNA-145 levels between stable angina and UA/NSTEMI were not found to be significantly different (p = 0.67).

[Figure omitted. See PDF.]

Fig 4. Plasma miRNA-145 levels and clinical presentation of CAD.

These bars represent averages of Ln_miRNA-145 values among different clinical presentations of CAD compared with non-CAD patients. T-bars indicate standard deviation. Patients with ST-elevation myocardial infarction had significantly lower values of Ln_miRNA-145 compared with the rest of the groups. Non-CAD subjects had significantly higher Ln_miRNA-145 than patients with CAD. * p <0.05; CAD, Coronary artery disease; RNA, Ribonucleic acid.

https://doi.org/10.1371/journal.pone.0123477.g004

Discussion

In summary, our study findings demonstrate that plasma levels of miRNA-145 are significantly lower in CAD patients compared with non-CAD subjects. These results reiterate findings from previous study that demonstrated significantly lower miRNA-145 levels in CAD patients [19]. Also, in our study we demonstrate that with increase in severity of CAD (multi-vessel disease and higher SYNTAX score) the levels are reduced further more. Within the CAD group, patients presenting with ACS, especially STEMI, had significantly lower miRNA-145 levels compared to other groups. However, it should also be noted that plasma miRNA-145 levels were not significantly different among stable angina and UA/NSTEMI patients.

Roles of miRNA-145 in atherosclerosis

VSMCs are thought to play a significant role in the pathogenesis of atherosclerosis. They are known to possess remarkable plasticity that allows them to reversibly modulate their phenotype during endothelial injury, inflammation and vessel wall injury [23]. Phenotype switching from quiescent ‘contractile’ or differentiated type to active ‘synthetic’ or dedifferentiated form is associated with enhanced proliferative rate, migration and formation of fibrous plaque [15,24]. In addition to the above ‘synthetic’ phenotype VSMCs also facilitate pathological lipid uptake and foam cell formation by enhanced expression of low-density lipoprotein, very low-density lipoprotein and scavenger receptors [23]. Circulating miRNA-145 has been shown to be regulated during coronary atherosclerosis [14,25]. Cheng Y et al [15] reported that miRNA-145 is the most abundant miRNA subtype in the normal vascular walls and that it is selectively expressed in the VSMCs of vessel wall, and can regulate the phenotype of VSMC. Experimental animal studies have reported that carotid artery ligation was associated with markedly reduced miRNA-145 levels, as measured by quantitative PCR and subsequent carotid stenosis [14]. Cordes and colleagues demonstrated that miRNA-145 promotes differentiation and represses proliferation of SMCs and the levels of this miRNA subtype were downregulated to nearly undetectable levels in atherosclerotic lesions containing neointimal hyperplasia [14]. Mechanisms by which miRNA-145 regulates VSMC phenotype include KLF4, calmodulin kinase IIδ, myocardin, actin polymerization and angiotensin-converting enzyme [26]. Restoration of miRNA-145 expression limits neointima formation in response to vascular injury by promoting KLF4 down regulation and VSMC contractile protein expression [15,26].

Circulating miRNA-145 as potential biomarkers for CAD

In this study, compared with that of controls, we demonstrated that plasma levels of miRNA-145 were significantly decreased in patients with CAD. These results suggest that downregulation of miRNA-145 plays a critical role in the pathogenesis of atherosclerotic plaques. Also, in the present study, miRNA-145 was correlated with multiple metabolic and CAD risk factors, including age, gender, diabetes mellitus, and tobacco use. Our study findings also show that miRNA-145 levels are associated negatively with the severity and extent of coronary stenotic lesions indicating further reduction in its plasma levels with increase in severity of CAD. CAD patients with multivessel disease, higher SYNTAX score and those with STEMI were associated with significantly lower miRNA-145 levels compared with others. To the best of our knowledge this is the first study to report on the association between plasma miRNA-145 levels and severity of CAD, especially patients with STEMI and high SYNTAX score, which further suggest that miRNA-145 might serve as biomarkers of CAD development and progression.

Several studies have in the past demonstrated the important of circulating microRNAs as potential biomarkers for CAD [27–29]. VSMCs play an important role in the stability and plaque rupture mechanisms [30]. While miRNA-145 can regulate vascular smooth muscle cell phenotype and extracellular matrix synthesis, thus possibly affecting the stability of the plaque, this article discusses the potential of plasma miRNA-145 as a clinical biomarker for the diagnosis and prognosis of acute coronary syndromes. However, it should be noted that using PCR to measure plasma miRNAs is expensive and time consuming, and there is a definite need to develop semi-automated techniques in the coming future to overcome this limitation.

MiRNA-145 as a potential therapeutic target for atherosclerosis

With the ability to regulate phenotypic differentiation of VSMCs, miRNA-145 can also act as a potential therapeutic target in the treatment of atherosclerosis [31,32]. Restoration of downregulated plasma miRNA-145 might inhibit neointimal lesion formation in the coronary arteries [14,32]. In a recent study by Lovren et al in experimental atherosclerosis it was demonstrated that targeted delivery of miRNA-145 into VSMCs in Apolipoprotein E knockout mice (ApoE-/-) was associated with markedly reduced plaque size, significantly increased fibrous cap area and plaque collagen content, and reduced necrotic core area at 12 weeks [32]. The study noted that apart from reducing overall atheroma burden, lentivirus mediated miRNA-145 transfer was associated with increased plaque stability thereby concluding that VSMC-specific miRNA-145 overexpression could be a novel therapeutic target in the management of atherosclerosis. However, further studies are warranted to establish miRNA-145 as a therapeutic option in atherosclerosis.

Limitations

Our study has several limitations. First, it is a single-center study involving a small sample size and therefore the results of the study should be interpreted with caution. Second, in our study we could not demonstrate the mechanisms behind the association between lower miRNA-145 levels in the plasma and CAD severity. Third, the distribution of patients in CAD and non-CAD groups was uneven. In our study we failed to demonstrate significant differences in miRNA-145 levels between patients with stable angina and unstable angina/non-ST-elevation myocardial infarction, and in patients with low and intermediate SYNTAX scores. Therefore our study results cannot be generalized to the whole CAD population. Large-scale multicenter studies are required to further elucidate the role of miRNA-145 as a potential marker in CAD, especially ACS. Finally, in our study we used C. elegans miRNA-39 as reference for normalization of miRNA-145 data. Some recent studies have reported using other housekeeping miRNAs for this purpose that might result in a different normalization data which might influence the overall results [33].

Conclusion

In conclusion, plasma miRNA-145 levels are significantly lower in CAD patients compared with those without significant CAD and this reduction seemed to positively correlate with the severity of CAD. These findings may have extensive clinical implications in the diagnosis and therapy of CAD.

Supporting Information

[Figure omitted. See PDF.]

S1 Table. Basic patient characteristics in the whole cohort.

https://doi.org/10.1371/journal.pone.0123477.s001

(DOCX)

Author Contributions

Conceived and designed the experiments: HG RRG JD MZ. Performed the experiments: HG LXS JD MZ. Analyzed the data: RRG YM LXS MZ. Contributed reagents/materials/analysis tools: HG LXS JD MZ. Wrote the paper: HG RRG YM LPL SPN DG MZ. Final approval of the manuscript submitted: HG RRG LPL DG JD MZ.

Citation: Gao H, Guddeti RR, Matsuzawa Y, Liu L-P, Su L-X, Guo D, et al. (2015) Plasma Levels of microRNA-145 Are Associated with Severity of Coronary Artery Disease. PLoS ONE 10(5): e0123477. https://doi.org/10.1371/journal.pone.0123477

References

1. Go AS, Mozaffarian D, Roger VL, Benjamin EJ, Berry JD, Blaha MJ et al. (2014) Heart disease and stroke statistics—2014 update: a report from the American Heart Association. Circulation 129: e28–e292. pmid:24352519

2. Yeh RW, Sidney S, Chandra M, Sorel M, Selby JV, Go AS. (2010) Population trends in the incidence and outcomes of acute myocardial infarction. N Engl J Med 362: 2155–2165. pmid:20558366

3. He J, Gu D, Wu X, Reynolds K, Duan X, Yao C et al. (2005) Major causes of death among men and women in China. N Engl J Med 353: 1124–1134. pmid:16162883

4. Ross R (1999) Atherosclerosis—an inflammatory disease. N Engl J Med 340: 115–126. pmid:9887164

5. Wei Y, Schober A, Weber C (2013) Pathogenic arterial remodeling: the good and bad of microRNAs. Am J Physiol Heart Circ Physiol 304: H1050–1059. pmid:23396454

6. Heusch G, Libby P, Gersh B, Yellon D, Bohm M, Lopaschuk G et al. (2014) Cardiovascular remodelling in coronary artery disease and heart failure. Lancet.

7. Orr AW, Hastings NE, Blackman BR, Wamhoff BR (2010) Complex regulation and function of the inflammatory smooth muscle cell phenotype in atherosclerosis. J Vasc Res 47: 168–180. pmid:19851078

8. Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116: 281–297. pmid:14744438

9. Sonkoly E, Stahle M, Pivarcsi A (2008) MicroRNAs and immunity: novel players in the regulation of normal immune function and inflammation. Semin Cancer Biol 18: 131–140. pmid:18291670

10. Schober A, Thum T, Zernecke A (2012) MicroRNAs in vascular biology—metabolism and atherosclerosis. Thromb Haemost 107: 603–604. pmid:22398634

11. Schroen B, Heymans S (2012) Small but smart—microRNAs in the centre of inflammatory processes during cardiovascular diseases, the metabolic syndrome, and ageing. Cardiovasc Res 93: 605–613. pmid:21994347

12. Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, Pogosova-Agadjanyan EL et al. (2008) Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci U S A 105: 10513–10518. pmid:18663219

13. Creemers EE, Tijsen AJ, Pinto YM (2012) Circulating microRNAs: novel biomarkers and extracellular communicators in cardiovascular disease? Circ Res 110: 483–495. pmid:22302755

14. Cordes KR, Sheehy NT, White MP, Berry EC, Morton SU, Muth AN et al. (2009) miR-145 and miR-143 regulate smooth muscle cell fate and plasticity. Nature 460: 705–710. pmid:19578358

15. Cheng Y, Liu X, Yang J, Lin Y, Xu DZ, Lu Q et al. (2009) MicroRNA-145, a novel smooth muscle cell phenotypic marker and modulator, controls vascular neointimal lesion formation. Circ Res 105: 158–166. pmid:19542014

16. Pleister A, Selemon H, Elton SM, Elton TS (2013) Circulating miRNAs: novel biomarkers of acute coronary syndrome? Biomark Med 7: 287–305. pmid:23547823

17. Libby P, Theroux P (2005) Pathophysiology of coronary artery disease. Circulation 111: 3481–3488. pmid:15983262

18. Kaudewitz D, Lee R, Willeit P, McGregor R, Markus HS, Kiechl S et al. (2013) Impact of intravenous heparin on quantification of circulating microRNAs in patients with coronary artery disease. Thromb Haemost 110: 609–615. pmid:23803974

19. Fichtlscherer S, De Rosa S, Fox H, Schwietz T, Fischer A, Liebetrau C et al. (2010) Circulating microRNAs in patients with coronary artery disease. Circ Res 107: 677–684. pmid:20595655

20. Fisher LD, Kennedy JW, Chaitman BR, Ryan TJ, McCabe C, Weiner D et al. (1981) Diagnostic quantification of CASS (coronary artery surgery study) clinical and exercise test results in determining presence and extent of coronary artery disease. A multivariate approach. Circulation 63: 987–1000. pmid:7471382

21. (1983) Coronary artery surgery study (CASS): a randomized trial of coronary artery bypass surgery. Survival data. Circulation 68: 939–950. pmid:6137292

22. Head SJ, Farooq V, Serruys PW, Kappetein AP (2014) The SYNTAX score and its clinical implications. Heart 100: 169–177. pmid:23539552

23. Doran AC, Meller N, McNamara CA (2008) Role of smooth muscle cells in the initiation and early progression of atherosclerosis. Arterioscler Thromb Vasc Biol 28: 812–819. pmid:18276911

24. Ishida M, Komori K, Yonemitsu Y, Taguchi K, Onohara T, Sugimachi K. (2001) Immunohistochemical phenotypic alterations of rabbit autologous vein grafts implanted under arterial circulation with or without poor distal runoff-implications of vein graft remodeling. Atherosclerosis 154: 345–354. pmid:11166767

25. Zhang C (2009) MicroRNA-145 in vascular smooth muscle cell biology: a new therapeutic target for vascular disease. Cell Cycle 8: 3469–3473. pmid:19829088

26. Albinsson S, Sessa WC (2011) Can microRNAs control vascular smooth muscle phenotypic modulation and the response to injury? Physiol Genomics 43: 529–533. pmid:20841497

27. Fichtlscherer S, Zeiher AM, Dimmeler S (2011) Circulating microRNAs: biomarkers or mediators of cardiovascular diseases? Arterioscler Thromb Vasc Biol 31: 2383–2390. pmid:22011751

28. Wang GK, Zhu JQ, Zhang JT, Li Q, Li Y, He J et al. (2010) Circulating microRNA: a novel potential biomarker for early diagnosis of acute myocardial infarction in humans. Eur Heart J 31: 659–666. pmid:20159880

29. Li C, Pei F, Zhu X, Duan DD, Zeng C (2012) Circulating microRNAs as novel and sensitive biomarkers of acute myocardial Infarction. Clin Biochem 45: 727–732. pmid:22713968

30. Johnson JL (2007) Matrix metalloproteinases: influence on smooth muscle cells and atherosclerotic plaque stability. Expert Rev Cardiovasc Ther 5: 265–282. pmid:17338671

31. Wei Y, Nazari-Jahantigh M, Neth P, Weber C, Schober A (2013) MicroRNA-126, -145, and -155: a therapeutic triad in atherosclerosis? Arterioscler Thromb Vasc Biol 33: 449–454. pmid:23324496

32. Lovren F, Pan Y, Quan A, Singh KK, Shukla PC, Gupta N et al. (2012) MicroRNA-145 targeted therapy reduces atherosclerosis. Circulation 126: S81–90. pmid:22965997

33. Sanders I, Holdenrieder S, Walgenbach-Brunagel G, von Ruecker A, Kristiansen G, Muller SC et al. (2012) Evaluation of reference genes for the analysis of serum miRNA in patients with prostate cancer, bladder cancer and renal cell carcinoma. Int J Urol 19: 1017–1025. pmid:22788411