Global cardiac-specific transgene expression using cardiopulmonary bypass with cardiac isolation
| Autor: | Stephane M. Konig, James M. Burkman, Hansell H. Stedman, Timothy J. Gardner, Alan S. Stewart, Mark M. Stecker, Terry Patterson, Haiyan Chen, Charles R. Bridges, Charles Yarnall, Ramin Malekan |
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| Rok vydání: | 2002 |
| Předmět: |
Pulmonary and Respiratory Medicine
medicine.medical_specialty Pathology Gene delivery law.invention Adenoviridae chemistry.chemical_compound Dogs law Internal medicine Albumins Cardiopulmonary bypass Medicine Myocyte Animals Transgenes Radionuclide Imaging Evans Blue Cardiopulmonary Bypass biology business.industry Myocardium Fissipedia Gene Transfer Techniques Heart biology.organism_classification medicine.disease chemistry Lac Operon Heart failure Circulatory system Cardiology Surgery Cardiology and Cardiovascular Medicine business Perfusion |
| Zdroj: | The Annals of thoracic surgery. 73(6) |
| ISSN: | 0003-4975 |
| Popis: | Background . The available techniques for intravascular gene delivery to the heart are inefficient and not organ-specific. Yet, effective treatment of heart failure will likely require transgene expression by the majority of cardiac myocytes. To address this problem, we developed a novel cannulation technique that achieves efficient isolation of the heart in situ using separate cardiopulmonary bypass (CPB) circuits for the heart and body in dogs. Methods . The arterial inflow and venous effluent from the two circuits were physically isolated. The efficiency of separation was 98% to 99% in three preliminary experiments using Evans Blue dye-labeled albumin. In 6 dogs, the cardiac circuit was perfused with oxygenated crystalloid cardioplegia at 37°C containing ≅ 4 × 10 11 particles of an adenovirus encoding LacZ (AdCMVLacZ) with a perfusion pressure of 170 to 200 mm Hg for 15 minutes allowing virus to recirculate through the heart ≅ 15 times. Cross-clamp time was 26 ± 2 minutes and CPB time was 90 ± 3 minutes. Results . Five animals survived and were euthanized at 7 days. β-Galactosidase activities measured using a chemiluminescent assay were three orders of magnitude higher in all areas of the heart than in the liver. Histological analyses revealed heterogeneous X-Gal staining of myocytes in all areas of the myocardium. Conclusions . Despite using a constitutive promoter, this technique yields relatively cardiac-specific transgene expression and is potentially translatable to clinical applications. Future studies will allow for further optimization of the conditions necessary for vector-mediated gene delivery to the heart. |
| Databáze: | OpenAIRE |
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