Microcirculation, 13: 187–197, 2006 Copyright c  2006 Taylor & Francis Group, LLC ISSN: 1073-9688 print / 1549-8719 online DOI: 10.1080/10739680600556811 Myocardial Gene Expression of Angiogenic Factors in Human Chronic Ischemic Myocardium: Influence of Acute Ischemia/Cardioplegia and Reperfusion YONGZHONG WANG, ∗ ANDERS GABRIELSEN, ‡ PATRICK R. LAWLER, ‡ GABRIELLE PAULSSON-BERNE, ‡ DANIEL A. STEINBR ¨ UCHEL, † GORAN K. HANSSON, ‡ AND JENS KASTRUP * ∗ Medical Department B, Cardiac Catheterization Laboratory, and † Department of Thoraxicsurgery, the Heart Centre, Copenhagen University Hospital, Rigshospitalet, Denmark; and ‡ Center for Molecular Medicine, Cardiovascular Research Unit, Karolinska Hospital, Stockholm, Sweden ABSTRACT Objective: Angiogenic therapies in animals have demonstrated the development of new blood vessels within ischemic myocardium. However, results from clinical protein and gene angiogenic trials have been less impressive. The present study aimed to investigate the expression of angiogenic genes in human chronic ischemic myocardium and the influence of acute ischemia/cardioplegia and reperfusion on their expression. Methods: Myocardial biopsies were taken from chronic ischemic and nonischemic myocardium in 15 patients with stable angina pectoris during coronary bypass surgery. Tissue samples were evaluated by oligonucleotide microarray and quantitative real-time PCR for the expression of angiogenic factors. Results: There was identical baseline expression of VEGF-A and VEGF-C mRNA in chronic ischemic myocardium compared with nonischemic myocardium. Reperfusion increased the gene expression of VEGF-A and VEGF-C mRNA both in nonischemic and ischemic myocardium. VEGF-A protein was detected mainly in the extracellular matrix around the cardiomyocytes in ischemic myocardium. Conclusion: These data suggest that the nonconclusive VEGF gene therapy trials chronic coronary artery disease was not due to a preexisting upregulation of VEGF in chronic ischemic myocardium. There might be room for further therapeutic angiogenesis in chronic ischemic myocardium. Microcirculation (2006) 13, 187–197. doi:10.1080/10739680600556811 KEY WORDS: angiogenesis, gene expression, growth factors, myocardial ischemia, reperfusion Angiogenesis and collateral blood vessel growth are adaptive responses to acute and chronic myocardial ischemia. Its purpose is to restore coronary blood flow and repair ischemic myocardium through the process of growing new blood vessels, which involves exten- sion of the preexisting vasculature by sprouting of new capillaries. However, spontaneous angiogenic re- sponse with development of collateral circulation is usually insufficient to reduce stress-induced ischemia in ischemic myocardium. One explanation may be the lack of adequate production of angiogenic factors. Address correspondence to Jens Kastrup, MD, DMSc, Medi- cal Department B, Cardiac Catheterization Laboratory 2014, the Heart Centre, Copenhagen University Hospital, Rigshospi- talet, 9 Blegdamsvej, DK-2100 Copenhagen, Denmark. E-mail: jkastrup@ rh.hosp.dk Received 24 June 2005; accepted 7 December 2005. Therapeutic angiogenesis by using vascular growth factor gene therapy may reduce myocardial ischemia by stimulating and augmenting the natural pro- cess of neovascularization. It is presently tested as an alternative treatment in patients with severe is- chemic heart disease (1–5). One of the most widely studied angiogenic factors in clinical trials is vas- cular endothelial growth factor (VEGF), a homod- imeric 34- to 46-kDa heparin-binding glycopro- tein. Intramyocardial gene transfer of VEGF initially demonstrated significant angiogenesis in preclinical studies. Small clinical trials have demonstrated that direct intramyocardial or intracoronary delivery of genes encoding VEGF seems to be safe and fea- sible (2–6). In addition, the studies showed clini- cal evidence of improved myocardial perfusion and thatVEGF may play a possible role in mobilization of bone marrow derived stem cells to the ischemic my- ocardium for vasculogenesis (2–6). However, larger