Vascular Endothelial Growth Factor Is Associated with the Morphologic and Functional Parameters in Patients with Hypertrophic Cardiomyopathy

Pudil, Radek; Vasatova, Martina; Fucikova, Alena; Rehulkova, Helena; Rehulka, Pavel; Palicka, Vladimir; Stulik, Jiri

doi:https://doi.org/10.1155/2015/762950

BioMed Research International

On this page

Abstract Introduction Methods Results Discussion Conclusions References Copyright Related Articles

Special Issue

Myocardial Diseases: Current Views on Etiopathogenesis, Diagnostic Modalities, and Therapeutic Options

View this Special Issue

Research Article | Open Access

Volume 2015 | Article ID 762950 | https://doi.org/10.1155/2015/762950

Vascular Endothelial Growth Factor Is Associated with the Morphologic and Functional Parameters in Patients with Hypertrophic Cardiomyopathy

Radek Pudil,¹Martina Vasatova,²Alena Fucikova,³Helena Rehulkova,³Pavel Rehulka,³Vladimir Palicka,²and Jiri Stulik³

Academic Editor: Giovanni Di Salvo

Received22 Oct 2015

Revised22 Nov 2015

Accepted10 Dec 2015

Published31 Dec 2015

Abstract

Background. Hypertrophic cardiomyopathy (HCM) is mostly autosomal dominant disease of the myocardium, which is characterized by myocardial hypertrophy. Vascular endothelial growth factor (VEGF) is involved in myocyte function, growth, and survival. The aim of study was to analyze the clinical significance of VEGF in structural and functional changes in patient with HCM. Methods. In a group of 21 patients with nonobstructive HCM, we assessed serum VEGF and analyzed its association with morphological and functional parameters. Compared to healthy controls, serum VEGF was increased: 199 (IQR: 120.4–260.8) ng/L versus 20 (IQR: 14.8–37.7) ng/L, . VEGF levels were associated with left atrium diameter (, ), left ventricle ejection fraction (, ), fractional shortening (, ), left ventricular mass (, ), LV mass index (, ), vena cava inferior diameter (, ), and peak gradient of tricuspid regurgitation (, ). Conclusions. Increased VEGF level is associated with structural and functional parameters in patients with HCM and serves as a potential tool for diagnostic process of these patients.

1. Introduction

Hypertrophic cardiomyopathy (HCM) is a heterogeneous myocardial disorder characterized by myocardial hypertrophy with structural and functional abnormalities. The incidence of HCM is approximately 1 in 500 (0.2%) of the general population [1]. The clinical outcome of HCM is diverse, ranging from asymptomatic patients to cardiac arrhythmias, congestive heart failure, and sudden cardiac death. According to the guidelines, patients with HCM require lifelong follow-up to detect changes in symptoms, risk of adverse events, left ventricle outflow tract obstruction (LVOTO), LV function, and cardiac rhythm [1, 2]. The recommended follow-up includes clinical evaluation, 12-lead ECG, and transthoracic echocardiography. The use of laboratory markers seems to be promising for diagnosis and risk stratification in HCM. There is a broad spectrum of biomarkers in peripheral blood, which are potentially useful for diagnosis and risk stratification in patients with HCM. But only cardiac troponins and natriuretic peptides have the most robust data [2]. The previous studies showed that high levels of natriuretic peptides and cardiac troponins are associated with cardiovascular events, heart failure, and death [3]. Systemic evaluation of these parameters revealed an association of these markers with the morphological and functional parameters [4–6]. The association of the myocardial hypertrophy and cardiac troponin and natriuretic peptide levels is not specific for the degree of left ventricle remodeling in HCM. The increase of these parameters is driven probably more by myocardial damage or pressure overload in patients with obstructive type of LV hypertrophy and not by hypertrophy per se.

Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen in vitro and angiogenic inducer in in vivo models. VEGF is secreted as a glycosylated homodimeric protein of 46 kDa that is made up of two 24 kDa subunits linked by disulphide bonds.

The tissue distribution of these VEGF receptors includes vascular smooth muscle cells, osteoblasts, cardiomyocytes, myofibroblasts, neurons, and various tumor cells [7]. It has been shown that VEGF is highly expressed in cardiomyocytes and myofibroblasts, indicating that VEGF family plays an autocrine/paracrine role in the regulation of myocyte and myofibroblast function and growth/survival [8–10]. Vascular endothelial growth factor plays an important role in the process of myocardial hypertrophy. However, the data about the association of VEGF and morphological and functional parameters in patients with hypertrophic cardiomyopathy are missing. The aim of our study was to assess VEGF level in peripheral blood in patients with HCM and to analyze its association with clinical markers of the disease.

2. Material and Methods

2.1. Study Population

Study population consisted of patients with nonobstructive hypertrophic cardiomyopathy. The diagnosis of hypertrophic cardiomyopathy was based on a history of illness, physical examination, echocardiography, and cardiac catheterization in accordance with European Society of Cardiology recommendations [2]. Based on the results of previous cardiac catheterization, patients with resting or provoked LVOTO were not included in the study. Also, patients with significant concomitant disease, such as pulmonary disease, arterial hypertension, malignancy, autoimmune disorders, neurodegenerative disorders, thyroid disease, or concurrent viral disease, were excluded. VEGF levels were compared to control group of healthy 20 blood donors ( years) with no evidence of cardiovascular disease according to ECG stress test and echocardiography. The study protocol conformed to the ethical guidelines of the 1975 Declaration of Helsinki and was approved by the ethical committee of our institution. Informed consent was obtained from each patient. Baseline demography and clinical characteristics of the study population are shown in Table 1.

2.2. Echocardiography

Echocardiography was performed in agreement with the American Society of Echocardiography and European Association of Cardiovascular Imaging standards evaluating the following parameters: left ventricular end-diastolic (LVEDD) and end-systolic (LVESD) diameters, left atrium diameter (LA), right ventricle diameter (RV), end-diastolic interventricular septum (IVST) and posterior wall thickness (PWT), inferior vena cava diameter (IVC), LV ejection fraction (LV EF), left ventricle fractional shortening (FS), peak gradient of tricuspid regurgitation (Pgrad TR), left ventricle mass (LVM), and left ventricle mass index (LVMI) [11]. Left ventricle outflow tract gradient was evaluated in accordance with European Society of Cardiology guidelines at rest and after provocation by Valsalva maneuver [2].

2.3. Assessment of VEGF

Blood samples were obtained from venous catheters, introduced into tube collectors containing no preservatives. Within 1 h, the blood samples were centrifuged for 10 min at 2500 g and the supernatant was removed and kept at −70°C until the assay was performed.

VEGF concentrations were measured using cytokine array for the evidence investigator protein biochip system (Randox Laboratories, UK). Simultaneous quantitative detection of multiple analytes based on sandwich chemiluminescent immunoassay was carried out from a single patient sample. The core technology consists of a solid plate containing discrete test regions with immobilized antibodies specific to different markers. Increased levels of marker in a specimen lead to increased binding of antibody labeled with horseradish peroxidase and thus to an increase in the luminescent signal emitted. The light signals generated from each of the test regions on the biochip are simultaneously detected using a charge coupled device camera. The analytical range of the VEGF assay was 3.24–1000 ng/L. The interassay coefficient of variability () was, for 146.4 ng/L, 10.8% and, for 456.3 ng/L, 7.4%. Internal quality control measurements were carried out using samples provided by the kit manufacturer.

2.4. Statistics

Statistical analysis was performed by MedCalc Software, version 14 (MedCalc Software bvba, Ostend, Belgium). Normally distributed variables are expressed as means ± standard deviation, while nonnormally distributed variables are expressed as median (interquartile range). Categorical variables are presented as percentages. Continuous variables were compared using Student’s -test, Mann-Whitney, or Wilcoxon’s tests, where appropriate. Linear regression was applied to evaluate the relationship between continuous variables. The degree of association between continuous variables was calculated using Pearson’s correlation coefficient. A value <0.05 was considered statistically significant.

3. Results

3.1. Serum VEGF

Serum VEGF was increased in all 21 patients. Compared with controls, HCM patients had significantly increased serum VEGF level 199 (IQR: 120.4–260.8) ng/L versus 20 (IQR: 14.8–37.7) ng/L, (Figure 1). In patients with New York Heart Association (NYHA) functional classes I and II, serum VEGF values were lower compared to patients with NYHA classes III and IV: 146.4 (IQR: 113.6–235.2) ng/L versus 328.1 (IQR: 286.7–406.3) ng/L, . In patients with atrial fibrillation, VEGF values were not significantly increased compared to patients with sinus rhythm: 234.1 (IQR: 132.2–307.5) ng/L versus 158.5 (IQR: 121.8–250.1) ng/L, 0.43.

3.2. Echocardiography

All patients underwent two-dimensional and Doppler echocardiography. Table 2 shows the summary information of echocardiographic parameters of patients with hypertrophic cardiomyopathy. The mean of the anteroposterior left atrium dimension was mm, and it exceeded reference values (female: 38 mm, male: 40 mm) in 17 (81%) patients. The mean internal diameter of right ventricle was mm, and it exceeded reference value (31 mm) in 1 (5%) patient. The mean of the internal end-systolic left ventricle dimension was mm, and it exceeded reference values (female: 34.8 mm, male: 39.8 mm) in 2 (10%) patients. The mean of the internal end-diastolic left ventricle dimension was mm, and it exceeded reference values (female: 52.2 mm, male: 58.4 mm) in 1 (5%) patient. The mean of the interventricular septum thickness was mm, and it exceeded recommended thickness for diagnosis of hypertrophic cardiomyopathy (≥15 mm) in all patients. The mean left ventricle mass was g, and it exceeded reference values for two-dimensional method (female: 150 g, male: 200 g) in all patients. The mean left ventricle mass index was g·m⁻², and it exceeded reference values for two-dimensional method (female: 88 g·m⁻², male: 102 g·m⁻²) in all patients. The mean of the left ventricle ejection fraction was , and, only in 1 (5%) patient, the LV ejection fraction was below the reference values (female: 54%, male: 52%). The mean of the left ventricle fractional shortening was , and it exceeded the reference values (female: 27–45, male: 25–43) in 3 (15%) patients. The mean peak tricuspid regurgitation gradient was mmHg. The inferior vena cava diameter was mm.

None of the patients had left ventricle outflow tract obstruction. The peak left ventricle outflow tract gradients were 6 (IQR: 1–7.5) mmHg during at rest measurement and 15 (IQR: 2.2–29) mmHg during Valsalva maneuver. The median values for the peak LVOT pressure gradient were 3 (IQR: 2.2–5) mmHg for at rest measurements and 14 (IQR: 2–22) mmHg during Valsalva maneuver.

3.3. Association of VEGF and Morphological and Functional Echocardiographic Parameters

The analysis of echocardiographic parameters showed association of the morphological and functional parameters with serum VEGF level. Serum levels of increased vascular endothelial growth factor left were significantly associated with left atrium diameter (, 95% CI: 0.09–0.77, and ), left ventricle ejection fraction (, 95% CI: −0.80–−0.17, and ), LV fractional shortening (, 95% CI: −0.79–−0.14, and ), LV mass (, 95% CI: 0.24–0.82, and ), LV mass index (, 95% CI: 0.02–0.75, and ), vena cava inferior diameter (, 95% CI: 0.30–0.84, and ), and peak gradient of tricuspid regurgitation (, 95% CI: 0.04–0.74, and ). Figures 2, 3, 4, and 5 illustrate regression analysis of the morphological and functional echocardiographic parameters.

(a)

(b)

(a)

(b)

(a)

(b)

Figure 4

Association of VEGF and left ventricle ejection fraction (a) and left ventricle fractional shortening (b) in patients with hypertrophic cardiomyopathy. Left ventricle ejection fraction (a): , coefficient of determination : 0.42, regression equation: , and significance level: . Left ventricle fractional shortening (b): , coefficient of determination : 0.27, regression equation: , and significance level: . LV EF, left ventricle ejection fraction; FS, left ventricular fractional shortening.

4. Discussion

Hypertrophic cardiomyopathy is an autosomal dominant inherited myocardial disease defined by the presence of increased left ventricular wall thickness that is not solely explained by abnormal loading conditions [2]. The pathophysiology of HCM is complex and consists of multiple interrelated abnormalities, including left ventricle hypertrophy, left ventricle outflow tract obstruction, diastolic dysfunction, mitral regurgitation, myocardial ischemia, and arrhythmias. The natural history of HCM varies from an asymptomatic and benign clinical course to sudden premature death. Therefore, the new markers are searched with the aim for detecting risk patients and improving their prognosis. Until now, there have been published many studies focused on a broad spectrum of biomarkers covering most of the pathophysiological processes described in HCM, for example, myocardial necrosis and wall stress markers, inflammatory markers, markers of endothelial dysfunction, markers of apoptosis, matrix metalloproteinases, platelet function markers, prothrombotic markers, hormones [12–14]. However, no disease specific marker was approved for a routine clinical practice.

In our study, we focused on VEGF and its clinical significance in patients with HCM.

It has been shown that VEGF is an endothelial cell-specific mitogen in vitro and an angiogenic inducer in in vivo models [15] and plays an important role in myocardial hypertrophy [16]. In our study, we confirmed increased serum VEGF level in patients with nonobstructive HCM when compared to normal population. Furthermore, VEGF levels were significantly associated with the degree of left ventricle hypertrophy (left ventricle mass and its index) in the absence of left ventricle outflow tract obstruction.

The progression of left ventricle hypertrophy leads to development of heart failure and is associated with a poor prognosis [17]. In various studies, plasma natriuretic peptide levels were significantly increased in HCM compared with normal subjects. In HCM patients, both BNP and ANP are significantly higher in the subgroup that shows evidence of obstruction and both correlate positively with left intraventricular pressure gradient. As prognostic factors, plasma levels of NT-pro-BNP and ANP are independent predictors of cardiovascular events in patients with HCM [17–20]. Therefore, we analyzed VEGF level and its relation to clinical and hemodynamic parameters. We observed that the New York Heart functional classes III and IV were characterized by higher VEGF level when compared to NYHA classes I and II. Hemodynamic parameters were evaluated by the use of echocardiography. We found significant negative association of left ventricular functional parameters (ejection fraction and fractional shortening). Also, we have noticed that VEGF is associated with increased pressure in pulmonary artery circulation. The increase of this pressure is passive and it is driven by volume and pressure left ventricle overload. This observation supports left atrium enlargement in the presence of LV hypertrophy. Our results are supported by the previous observations of the pathogenetic role of the VEGF in left ventricle remodeling process in patients with HCM [21–23]. As compared to other markers, VEGF could have a potential to reflect morphological and functional parameters in patients with hypertrophic cardiomyopathy.

5. Conclusions

The results of our present study indicate that serum VEGF level in patients with HCM was significantly increased in comparison to control group and was associated with NYHA functional class. The VEGF levels were positively associated with the morphological parameters of HCM (left ventricle mass and its index); functional parameters of the left ventricle showed negative association. Serum VEGF correlated also with peak tricuspid regurgitation gradient (marker of pulmonary circulation pressure), inferior vena cava diameter, and left atrium diameter (markers of volume and pressure overload).

Despite the study limitations (relatively small number of the patients and morphological and hemodynamic parameters were evaluated by echocardiography), serum VEGF has emerged as an interesting and promising parameter useful for risk stratification of patients with hypertrophic cardiomyopathy.

Conflict of Interests

The authors declare that there is no conflict of interests regarding the publication of this paper.

Acknowledgment

This work was supported by a grant from Ministry of Health of the Czech Republic no. NT 13721.

References

B. J. Gersh, B. J. Maron, R. O. Bonow et al., “2011 ACCF/AHA Guideline for the diagnosis and treatment of hypertrophic cardiomyopathy: a report of the American College of Cardiology Foundation/American Heart Association Task Force on Practice Guidelines developed in collaboration with the American Association for Thoracic Surgery, American Society of Echocardiography, American Society of Nuclear Cardiology, Heart Failure Society of America, Heart Rhythm Society, Society for Cardiovascular Angiography and Interventions, and Society of Thoracic Surgeons,” Journal of the American College of Cardiology, vol. 58, no. 25, pp. e212–e260, 2011.
View at: Publisher Site | Google Scholar
P. M. Elliott, A. Anastasakis, M. A. Borger et al., “2014 ESC Guidelines on diagnosis and management of hypertrophic cardiomyopathy: the Task Force for the Diagnosis and Management of Hypertrophic Cardiomyopathy of the European Society of Cardiology (ESC),” European Heart Journal, vol. 35, no. 39, pp. 2733–2779, 2014.
View at: Publisher Site | Google Scholar
J. B. Geske, P. M. McKie, S. R. Ommen, and P. Sorajja, “B-type natriuretic peptide and survival in hypertrophic cardiomyopathy,” Journal of the American College of Cardiology, vol. 61, no. 24, pp. 2456–2460, 2013.
View at: Publisher Site | Google Scholar
C. J. Coats, M. J. Gallagher, M. Foley et al., “Relation between serum N-terminal pro-brain natriuretic peptide and prognosis in patients with hypertrophic cardiomyopathy,” European Heart Journal, vol. 34, no. 32, pp. 2529–2537, 2013.
View at: Publisher Site | Google Scholar
T. Kubo, H. Kitaoka, M. Okawa et al., “Serum cardiac troponin I is related to increased left ventricular wall thickness, left ventricular dysfunction, and male gender in hypertrophic cardiomyopathy,” Clinical Cardiology, vol. 33, no. 2, pp. E1–E7, 2010.
View at: Publisher Site | Google Scholar
T. Kubo, H. Kitaoka, S. Yamanaka et al., “Significance of high-sensitivity cardiac troponin T in hypertrophic cardiomyopathy,” Journal of the American College of Cardiology, vol. 62, no. 14, pp. 1252–1259, 2013.
View at: Publisher Site | Google Scholar
M. Shibuya, “Vascular endothelial growth factor and its receptor system: physiological functions in angiogenesis and pathological roles in various diseases,” Journal of Biochemistry, vol. 153, no. 1, pp. 13–19, 2013.
View at: Publisher Site | Google Scholar
T. Zhao, W. Zhao, W. Meng et al., “VEGF-C/VEGFR-3 pathway promotes myocyte hypertrophy and survival in the infarcted myocardium,” American Journal of Translational Research, vol. 7, no. 4, pp. 697–709, 2015.
View at: Google Scholar
F. Cambronero, F. Marín, V. Roldán, D. Hernández-Romero, M. Valdés, and G. Y. H. Lip, “Biomarkers of pathophysiology in hypertrophic cardiomyopathy: implications for clinical management and prognosis,” European Heart Journal, vol. 30, no. 2, pp. 139–151, 2009.
View at: Publisher Site | Google Scholar
E. Kaza, K. Ablasser, D. Poutias et al., “Up-regulation of soluble vascular endothelial growth factor receptor-1 prevents angiogenesis in hypertrophied myocardium,” Cardiovascular Research, vol. 89, no. 2, pp. 410–418, 2011.
View at: Publisher Site | Google Scholar
R. M. Lang, L. P. Badano, V. Mor-Avi et al., “Recommendations for cardiac chamber quantification by echocardiography in adults: an update from the American Society of Echocardiography and the European Association of Cardiovascular Imaging,” European Heart Journal—Cardiovascular Imaging, vol. 16, no. 3, pp. 233–271, 2015.
View at: Publisher Site | Google Scholar
B. A. Houston and G. R. Stevens, “Hypertrophic cardiomyopathy: a review,” Clinical Medicine Insights: Cardiology, vol. 8, supplement 1, pp. 53–65, 2014.
View at: Google Scholar
B. N. Putko, Z. Wang, J. Lo et al., “Circulating levels of tumor necrosis factor-alpha receptor 2 are increased in heart failure with preserved ejection fraction relative to heart failure with reduced ejection fraction: evidence for a divergence in pathophysiology,” PLoS ONE, vol. 9, no. 6, Article ID e99495, 2014.
View at: Publisher Site | Google Scholar
C. Roma-Rodrigues, L. R. Raposo, and A. R. Fernandes, “MicroRNAs based therapy of hypertrophic cardiomyopathy: the road traveled so far,” BioMed Research International, vol. 2015, Article ID 983290, 8 pages, 2015.
View at: Publisher Site | Google Scholar
N. Ferrara, “Vascular endothelial growth factor: basic science and clinical progress,” Endocrine Reviews, vol. 25, no. 4, pp. 581–611, 2004.
View at: Publisher Site | Google Scholar
J. Alkon, M. K. Friedberg, C. Manlhiot et al., “Genetic variations in hypoxia response genes influence hypertrophic cardiomyopathy phenotype,” Pediatric Research, vol. 72, no. 6, pp. 583–592, 2012.
View at: Publisher Site | Google Scholar
B. J. Maron, V. N. Tholakanahalli, A. G. Zenovich et al., “Usefulness of B-type natriuretic peptide assay in the assessment of symptomatic state in hypertrophic cardiomyopathy,” Circulation, vol. 109, no. 8, pp. 984–989, 2004.
View at: Publisher Site | Google Scholar
J. Magga, P. Sipola, O. Vuolteenaho et al., “Significance of plasma levels of N-terminal pro-B-type natriuretic peptide on left ventricular remodeling in non-obstructive hypertrophic cardiomyopathy attributable to the Asp175Asn mutation in the α-tropomyosin gene,” American Journal of Cardiology, vol. 101, no. 8, pp. 1185–1190, 2008.
View at: Publisher Site | Google Scholar
M. Okawa, H. Kitaoka, Y. Matsumura et al., “Functional assessment by myocardial performance index (Tei index) correlates with plasma brain natriuretic peptide concentration in patients with hypertrophic cardiomyopathy,” Circulation Journal, vol. 69, no. 8, pp. 951–957, 2005.
View at: Publisher Site | Google Scholar
H. Matsuura, T. Murakami, K. Hina et al., “Association of elevated plasma B-type natriuretic peptide levels with paroxysmal atrial fibrillation in patients with nonobstructive hypertrophic cardiomyopathy,” Clinical Biochemistry, vol. 41, no. 3, pp. 134–139, 2008.
View at: Publisher Site | Google Scholar
I. Friehs, R. Barillas, N. V. Vasilyev, N. Roy, F. X. McGowan, and P. J. Del Nido, “Vascular endothelial growth factor prevents apoptosis and preserves contractile function in hypertrophied infant heart,” Circulation, vol. 114, no. 1, supplement, pp. I290–I295, 2006.
View at: Publisher Site | Google Scholar
A. Leychenko, E. Konorev, M. Jijiwa, and M. L. Matter, “Stretch-Induced hypertrophy activates NFkB-Mediated VEGF secretion in adult cardiomyocytes,” PLoS ONE, vol. 6, no. 12, Article ID e29055, 2011.
View at: Publisher Site | Google Scholar
N. Shimojo, S. Jesmin, S. Zaedi et al., “Contributory role of VEGF overexpression in endothelin-1-induced cardiomyocyte hypertrophy,” American Journal of Physiology—Heart and Circulatory Physiology, vol. 293, no. 1, pp. H474–H481, 2007.
View at: Publisher Site | Google Scholar

Copyright

Copyright © 2015 Radek Pudil et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PDF Download Citation

Download other formats

Order printed copies

Views

1196

Downloads

994

Citations