title: DNA microarray analysis for human congenital heart disease.
abstract: 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 immuno histochemistry, 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 Ialpha, 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.
authors.affiliation: Department of Pharmacology, Erasmus MC, University Medical Center, Rotterdam, The Netherlands. h.sharma@erasmusmc.nl
authors.forename: Hari S
authors.initials: HS
authors.lastname: Sharma
authors.forename: Theodorus H F
authors.initials: TH
authors.lastname: Peters
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Moorhouse
authors.forename: Peter J
authors.initials: PJ
authors.lastname: van der Spek
authors.forename: Ad J J C
authors.initials: AJ
authors.lastname: Bogers
mesh_terms: D000328:Adult; D002318:Cardiovascular Diseases / Q000235:genetics / Q000473:pathology; D019295:Computational Biology; D015536:Down-Regulation / Q000235:genetics; D004719:Endomyocardial Fibrosis / Q000235:genetics / Q000473:pathology; D016326:Extracellular Matrix Proteins / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D020869:Gene Expression Profiling; D023281:Genomics; D006330:Heart Defects, Congenital / Q000235:genetics* / Q000473:pathology; D006801:Humans; D017380:Hypertrophy, Right Ventricular / Q000235:genetics / Q000473:pathology; D007223:Infant; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D013771:Tetralogy of Fallot / Q000235:genetics / Q000473:pathology; D015854:Up-Regulation / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics
chemical_list: D016326:Extracellular Matrix Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2