DNA microarray analysis for human congenital heart disease
Right ventricular hypertrophy and failure are prominent features in cyanotic congenital heart disease, tetralogy of Fallot (TF). Patients with TF require primary cardiac surgery at a very young age. To gain insight into the underlying molecular mechanisms of right ventricular hypertrophy and to identify gene(s) involved in TF, differential gene expression profile was assessed using expression-based microarray technology on right ventricular biopsies from young TF patients who underwent primary correction. By using quantitative immunohistochemistry, expression of vascular endothelial growth factor (VEGF), flk-1, and extracellular matrix (ECM) proteins (collagens and fibronectin) as well as vessel counts and myocyte cell size was evaluated in TF patients in relation to age-matched controls. Among 236 genes showing altered expression pattern in TF patients, VEGF (1.8-fold) and ECM markers were clearly upregulated (fibronectin, 2.4-fold; collagen Iα, 7.5-fold; and collagen III, 4.4-fold); flk-1 and most matrix metalloproteinases (MMPs) remained unchanged, except the levels of MMP-13 and-17 declined. Tissue inhibitors of metalloproteinases showed a downregulated pattern. Staining of VEGF in cardiomyocytes and of ECM proteins (fibronectin, collagen I and III) in interstitial as well as in perivascular area was increased (p<0.01) in TF patients. Morphometric analysis revealed enhanced vascular density (p<0.05) with unchanged wall thickness and enlarged myocyte cross-sectional areas (p<0.01) with linear correlation (p<0.01) with the age in TF-1 patients. We conclude that the upregulation of genes encoding VEGF and ECM proteins are the key events contributing to right ventricular hypertrophy and stunted angiogenesis in patients with TF.
Index EntriesTetralogy of Fallot right ventricular hypertrophy DNA microarray fibrosis angiogenesis video image analysis
Srivastava, D. (2004) Heart disease: an ongoing genetic battle? Nature
, 819–822.PubMedCrossRefGoogle Scholar
Nediani, C., Formigli, L., Perna, A. M., et al. (2000) Early changes induced in the left ventricle by pressure overload: an experimental study on swine heart. J. Mol. Cell. Cardiol.
, 131–142.PubMedCrossRefGoogle Scholar
Bauer, E. P., Kuki, S., Zimmermann, R., et al. (1998) Upregulated and downregulated transcription of myocardial genes after pulmonary artery banding in pigs. Ann. Thorac. Surg.
, 527–531.PubMedCrossRefGoogle Scholar
Pigula, F. A., Khalil, P. N., Mayer, J. E., et al. (1999) Repair of tetralogy of Fallot in neonates and young infants. Circulation
, II157-II161.PubMedGoogle Scholar
Van Arsdell, G. S., Maharaj, G. S., Tom, J., et al. (2000) What is the optimal age for repair of tetralogy of Fallot? Circulation
, III123-II129.PubMedGoogle Scholar
Seliem, M. A., Wu, Y. T., and Glenwright, K. (1995) Relation between age at surgery and regression of right ventricular hypertrophy in tetralogy of Fallot. Pediatr. Cardiol.
, 53–55.PubMedCrossRefGoogle Scholar
Schwartz, S. M., Gordon, D., Mosca, R. S., et al. (1996) Collagen content in normal, pressure, and pressure-volume overloaded developing human hearts. Am. J. Cardiol.
, 734–738.PubMedCrossRefGoogle Scholar
Konstantinov, I. E., Coles, J. G., Boscarino, C., et al. (2004) Gene expression profiles in children undergoing cardiac surgery for right heart obstructive lesions. J. Thorac. Cardiovasc. Surg.
, 746–754.PubMedCrossRefGoogle Scholar
Kruse, J. J., te Poele, J. A., Russell, N. S., Boersma, L. J., and Stewart, F. A. (2004). Microarray analysis to identify molecular mechanisms of radiation-induced microvascular damage in normal tissues. Int. J. Radiat. Oncol. Biol. Phys.
, 420–426.PubMedCrossRefGoogle Scholar
Dempsey, A. A., Dzau, V. J., and Liew, C. C. (2001) Cardiovascular genomics: estimating the total number of genes expressed in the human cardiovascular system. J. Mol. Cell. Cardiol.
, 1879–1886.PubMedCrossRefGoogle Scholar
Peng, C. F., Wei, Y., Levsky, J. M., et al. (2002) Microarray analysis of global changes in gene expression during cardiac myocyte differentiation. Physiol. Genomics
, 145–155.PubMedGoogle Scholar
Zhao, X. S., Gallardo, T. D., Lin, L., et al. (2002) Transcriptional mapping and genomic analysis of the cardiac atria and ventricles. Physiol. Genomics
, 53–60.PubMedGoogle Scholar
Cook, S. A., Matsui, T., Li, L., et al. (2002) Transcriptional effects of chronic Akt activation in the heart. J. Biol. Chem.
, 22,528–22,533.Google Scholar
Steenbergen, C., Afshari, C. A., Petranka, J. G., et al. (2003) Alterations in apoptotic signaling in human idiopathic cardiomyopathic hearts in failure. Am. J. Physiol.
, H268-H276.Google Scholar
Yussman, M. G., Toyokawa, T., Odley, A., et al. (2002) Mitochondrial death protein Nix induced in cardiac hypertrophy and triggers apoptotic cardiomyopathy. Nat. Med.
, 725–730.PubMedGoogle Scholar
Stanton, L. W., Garrard, L. J., Damm, D., et al. (2000) Altered patterns of gene expression in response to myocardial infarction. Circ. Res.
, 939–945.PubMedGoogle Scholar
Friddle, C. J., Koga, T., Rubin, E. M., et al. (2000) Expression profiling reveals distinct sets of genes altered during induction and regression of cardiac hypertrophy. Proc. Natl. Acad. Sci. USA
, 6745–6750.PubMedCrossRefGoogle Scholar
Barrans, J. D., Stamatiou, D., and Liew, C. (2001) Construction of a human cardiovascular cDNA microarray: portrait of the failing heart. Biochem. Biophys. Res. Commun.
, 964–969.PubMedCrossRefGoogle Scholar
Yang, J., Moravec, C. S., Sussman, M. A., et al. (2000) Expression profiling reveals distinct sets of genes altered during induction and regression of cardiac hypertrophy. Proc. Natl. Acad. Sci. USA
, 6745–6750.CrossRefGoogle Scholar
Chien, K. R. (2000) Genomic circuits and the integrative biology of cardiac diseases. Nature
, 227–232.PubMedCrossRefGoogle Scholar
Sharma, H. S., van Heugten, H. A., Goedbloed, M. A., et al. (1994) Angiotensin II induced expression of transcription factors precedes increase in transforming growth factor-β1 mRNA in neonatal cardiac fibroblasts. Biochem. Biophys. Res. Commun.
, 105.PubMedCrossRefGoogle Scholar
Brand, T., Sharma, H. S., Fleischmann, K. E., et al. (1992) Proto-oncogene expression in porcine myocardium subjected to ischemia and reperfusion. Circ. Res.
, 1351–1360.PubMedGoogle Scholar
Tan, F. L., Moravec, C. S., Li, J. et al. (200) The gene expression fingerprint of human heart failure. Proc. Natl. Acad. Sci. USA
, 11,387–11392.Google Scholar
Yajima, N., Masuda, M., Miyazaki, M., et al. (2002) Oxidative stress is involved in the development of experimental abdominal aortic aneurysm: a study of the transcription profile with complementary DNA microarray. J. Vasc. Surg.
, 379–385.PubMedCrossRefGoogle Scholar
Peters, T. H. F., Sharma, H. S., Yilmaz, E., et al. (1999) Quantitative analysis of collagens and fibronectin expression in human right ventricular hypertrophy. Ann. N.Y. Acad. Sci.
, 278–285.PubMedCrossRefGoogle Scholar
Peters, T. H. F., Sharma, H. S., and Bogers, A. J. J. C. (2003) Computerized image analysis in the quantitative assessment of interstitial fibrosis late after correction of tetralogy of Fallot. Cardiovas. Eng.
, 114–120.Google Scholar
Peters, T. H. F., de Jong, P. L., Klompe, L., et al. (2003) Right ventricular collagen and fibronectin levels in patients with pulmonary atresia and ventricular septal defect. Mol. Cell. Biochem.
, 27–32.PubMedCrossRefGoogle Scholar
Bishop, J. E., Rhodes, S., Laurent, G. J., et al. (1994) Increased collagen synthesis and decreased collagen degradation in right ventricular hypertrophy induced by pressure overload. Cardiovasc. Res.
, 1581–1585.PubMedCrossRefGoogle Scholar
Spinale, F. G. (2002) Matrix metalloproteinases: regulation and dysregulation in the failing heart. Circ. Res.
, 520–530.PubMedCrossRefGoogle Scholar
Vikstrom, K. L., Bohlmeyer, T., Factor, S. M., et al. (1998) Hypertrophy, pathology, and molecular markers of cardiac pathogenesis. Circ. Res.
, 773–778.PubMedGoogle Scholar