The American Journal of Cardiology© OCTOBER 16-21, 2005 TCT ABSTRACTS/Oral 9H

ORAL

ABSTRACTS

Cardiovascular Genetics, Genomics, and ProteomicsRoom 151AB

Monday, October 17, 2005, 3:30 pm - 4:30 pm(Abstract nos. 17-20)

TCT-17

Recombinant Adeno-Associated Virus Serotype 1 Vector-Mediated the Delivery of VEGF Gene in Mouse Ischemic Skeletal MusclesHua Yan1, Xiaobing Wu2, Yanhong Guo1, Wei Gao1 1Peking University Third Hospital, Beijing, China2AGTC Gene Technology Company Ltd., Beijing, China

Background: The serotypes of adeno- sociated virus (AAV) have the potential to become important resources for clinical gene therapy. Recent studies have compared the serotype-specific virion shells on vectors transduction in different tissues and suggested rAAV2 b ed vectors pseudotyped with serotype 1 capsid proteins (rAAV1 vectors) allow superior gene transfer into normal skeletal muscles. Method and results: To evaluate the potential role of rAAV1 vector gene transfer in the treatment of skeletal muscle ischemia, 3 × 109

vector genomes (vg), 3 × 1010 vg and 3 × 1011 vg rAAV1-VEGF165 vectors for each mouse were injected in the mouse ishcemic skeletal muscles. rAAV1-LacZ injected mice were controls. We demonstrated that VEGF _expression in different groups were dosage- dependent me ured by ELISA at 1 month post vector injection. Both 3 × 1010 vg and 3 × 1011 vg rAAV1-VEGF165 vectors for each mouse were sufficient for angiogenesis and arteriogenesis in ischmic skeletal muscles. Furthermore, the newly formed vessels in muscles injected with 3 × 1010 vg of the rAAV1-VEGF165 vectors were significantly less leaky compared to those in muscles injected with 3 × 1011 vg of the rAAV1-VEGF165 vectors. Conclusion: Our results suggest that rAAV1-VEGF165 vector may be a useful tool in the treatment of muscle ischemia and a lower rAAV1-VEGF165 vector dosage is sufficient for new vessel genesis

TCT-18

Prospective Evaluation of the Effect of Paraoxonase Gln-Arg 192 Polymorphism on the Long Term Risk of Major Adverse Cardiac Events After Coronary StentingKaram Souibri1, Adolphe Shayne1, Emmanuelle Filippi-Codaccioni1, Sabine Fradin2, Rémi Morello3, Gilles Grollier1, Martial Hamon1 1Service de Cardiologie, Caen, France2Service de Biochimie, Caen, France3Service de Biostatistiques, Caen, France

Background: Oxidative modification of low density lipoproteins (ox-LDL) in the artery wall is currently believed to be central in the pathogenesis of atherosclerosis. Paraoxonase (PON1) is a high density lipoprotein (HDL) associated enzyme that protects both HDL and LDL against oxidation. Objective: We sought to evaluate the association between Gln (Q)-Arg (R) 192 polymorphism of PON1 gene and the risk of major adverse cardiac events (MACE) in patients with established coronary artery disease (CAD) treated by PCI. Methods: From January 1997, to January 1999, 1456 consecutive patients have had stents in at least one of their coronary arteries. We used blood samples for genotypic study after the procedure and the patients’ written consent. 203 DNA samples couldn’t be used, and 11 patients were lost during the 5.7 mean years follow-up period, which brought the final cohort to 1222 patients. Long term clinical outcome was obtained and the rates of MACE (death, acute myocardial infarction, percutaneous or surgical myocardial revascularization ) were compared according to the insertion/deletion (Q/R) polymorphism of PON1. Results: Of the 1222 patients, 46.3% had the QQ genotype, 43.6 had the QR

genotype and 9.9% had the RR genotype. All baseline clinical, angiographic and procedural characteristics were identical in the three groups. Event-free survival during the follow-up period was identical in patients with QQ genotype compared to those carrying one or two R alleles. The predictors of cardiovascular death in patients with CAD were age >= 65 years (RR=2, CI 0.95-4.19, p=0.068), creatinine clearance <80ml/min (RR=3.72, CI 1.36-10.23, p=0.011), and LVEF<40% (RR=2.68, CI 1.45-4.97, p=0.002). There is no difference for the other MACE according to PON1 polymorphism. Conclusion: In this large prospective study PON1 Gln-Arg 192 polymorphism does not influence the long term prognosis of stented patients. Our results do not discard definitely the pathogenic role of PON1 polymorphism in CAD but indicates that screening these patients for this gene is not useful for secondary prevention strategies. PON1 activity and concentration rather than its polymorphism may identify patients at risk for MACE.

TCT-19

Nitric Oxide Synthase-3 Polymorphism and Outcomes after Percutaneous Coronary InterventionA.J. Conrad Smith, Oscar C. Marroquin, Suresh R. Mulukutla, Pratik B. Patel, William D. Anderson, Joon Sup Lee, Debra Rosenfelder, Karen Janosko, Dennis M. McNamara University of Pittsburgh Medical Center, Pittsburgh, PA

Introduction: Nitric oxide influences platelet function and smooth muscle cell proliferation after percutaneous coronary intervention (PCI). A polymorphism is present in endothelial nitric oxide synthase (NOS3), Asp298Glu. The Asp variant is a risk factor for CAD, however the impact on outcomes post PCI is uncertain. We evaluated the NOS3 polymorphism and revascularization events the first year after PCI Method: 674 patients referred for PCI were enrolled in a registry from January 2000 until 2003. Most PCI were with bare metal stents. Subjects were genotyped and followed for one year or until a repeat revascularization event (PCI or CABG). Kaplan-Meier event-free survival was compared by genotype subset. Results: The cohort (mean age 64+ 11) was 96% white and 67% male. In terms of NOS3, 40% of subjects were Glu298Glu, 49% were Glu298Asp heterozygotes, and 11% were Asp298Asp homozygotes. During the course of follow-up, 138 events occurred: 89 repeat PCI (13%), 37 CABG (6%), and 12 deaths(2%). The overall event-free survival at 30, 90,180 and 365 days was 98, 92, 87 and 80%. Asp298 was associated with poorer event free survival (Figure, p=0.008). Event-free survival was poorest in Asp298Asp (96,88,81,69%) intermediate in heterozygotes and best in Glu298Glu subjects (99,95,88, 84%). Conclusion: Subjects with NOS3 variantAsp298 have poorer event free survival after PCI. Analysis of genetic background has the potential to delineate populations at greater risk for repeat intervention.

MONDAY 10/17/05 3:30 PM-4:30 PM (Room 151AB)

10H The American Journal of Cardiology© OCTOBER 16-21, 2005 TCT ABSTRACTS/Oral

ORAL

ABSTRACTS

TCT-20

Cellular and Humeral Characteristics of Injected Bone Marrow cells Fail to Predict Clinical Response in Patients with Advanced Coronary Artery DiseaseShmuel Fuchs1, Richard Baffour2, Giora Weisz3, Petros Okubagzi4, Alexander Battler1, Martin B Leon3, Stephen E Epstein2, Ran Kornowski1 1Rabin Medical Center, Petach Tikva, Israel2Cardiovascular Research Institute, Washington, DC;3Cardiovascular Research Foundation, New York, NY;4Cardiovascualr Research Institute, Washington, DC

Background: Intra-myocardial injection of autologous bone marrow (BM) may improve clinical response in “no option” patients with refractory angina and myocardial ischemia. The improvement, however, is not uniform and variability in clinical response was noted. We sought to assess whether cellular and humeral characteristics of the injected BM cells predict clinical response. Methods: BM cells obtained from 27 patients were isolated by Ficoll-Hypaque gradient centrifugation and cultured in long-term culture medium (LTCM). Weekly, half of the medium was removed and replaced with fresh LTCM. The concentration of VEGF and MCP-1 in conditioned medium was measured using ELISA. The BM cell population was analyzed by FACS analyzer using laser light side scattering and monoclonal antibodies against CD34 and CD45. Results: At 3 months, exercise duration time improved in 16 patients (2.5±2.0 min) and failed to improve in 11 (-1.0±1.1 min). The number of total injected nucleated cells (CD45+), progenitor cells (CD34+) or the magnitude of ex-vivo secretion VEGF and MCP-1 by cultured BM cells failed to predict change in exercise response (Table). Similar results were obtained when CCS class was used as the response criteria (21 responders, data not shown). Conclusions: Cellular and humeral characteristics of ABM cells do not predict clinical response following transendocardial injection of these cells in patients with refractory angina, underscoring the advisability for further mechanistic exploration.

Responders Non-Responders PTotal injected cells (x106) 25±27 28±30 0.85CD34+ cell number (x106) 0.72±1.85 0.38±0.28 0.33VEGF (at week 4, ng/ml) 4,975±3704 3,093±1467 0.70ΔVEGF (between week 4 and 1, ng/ml)

4,496±3185 2,921±1393 0.75

MCP-1 (at week 4, ng/ml 2,178±295 2,317±379 0.10ΔMCP-1 (between week 4 and 1, ng/ml)

1,002±516 943±627 0.8

MONDAY 10/17/05 3:30 PM-4:30 PM (Room 151AB)