Abstract | OBJECTIVE: To confirm gene transfer techniques especially into the whole heart, we tried out a gene transfer method involving liposome with the viral envelope hemagglutinating virus of Japan liposome as an alternative to existing techniques such as cationic lipofection or other viral vectors. METHOD: RESULTS:
Fluorescein isothiocyanate was detected in the nuclei of more than 70% of the myocytes (75% +/- 14%, n = 5) in the H group compared with fewer than 10% in the L group (7% +/- 5%, n = 5). The intensity of fluorescein isothiocyanate was significantly higher in the H group (979 +/- 112 FI) than in the L group (116 +/- 68 FI). beta-Galactosidase was expressed in the cytosol of more than 50% of the myocytes in the H group (61% +/- 7%, n = 5) compared with none in the L group (0%, n = 5). After 3 days of gene transfection, and when exposed to ischemia (30 minutes, 37 degrees C) and reperfusion (30 minutes, 37 degrees C) with Langendorff apparatus, the hearts transfected with manganese-superoxide dismutase (S group, n = 5) showed a significantly higher percentage of recovery of left ventricular end-diastolic pressure (S vs C, 86% +/- 3% vs 54% +/- 12%) and coronary flow (98% +/- 2% vs 66% +/- 12%) than did the control hearts (C group, n = 5). Western blotting analysis showed an apparent increased expression of manganese-superoxide dismutase in the hearts transfected with manganese-superoxide dismutase compared with the control hearts. These results clearly demonstrated that the donor hearts were transfected with fluorescein-labeled oligonucleotide and the beta-galactosidase gene as a result of coronary infusion of the hemagglutinating virus of Japan liposome during cardioplegic arrest at the time of harvest. Furthermore, the hearts transfected with manganese-superoxide dismutase showed significant improvement in tolerance against ischemia reperfusion injury. CONCLUSION: We believe that this method represents a novel in vivo gene transfer technique for the heart and thus may provide a new tool for research and therapy of heart transplantation.
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Authors | Y Sawa, K Kadoba, K Suzuki, H Z Bai, Y Kaneda, R Shirakura, H Matsuda |
Journal | The Journal of thoracic and cardiovascular surgery
(J Thorac Cardiovasc Surg)
Vol. 113
Issue 3
Pg. 512-8; discussion 518-9
(Mar 1997)
ISSN: 0022-5223 [Print] United States |
PMID | 9081096
(Publication Type: Journal Article)
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Chemical References |
- Liposomes
- Oligonucleotides
- Manganese
- Superoxide Dismutase
- beta-Galactosidase
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Topics |
- Animals
- Coronary Vessels
- Gene Transfer Techniques
- Genetic Vectors
- Heart Arrest, Induced
- Heart Transplantation
(physiology)
- Liposomes
- Manganese
- Oligonucleotides
- Rats
- Rats, Sprague-Dawley
- Respirovirus
(genetics)
- Superoxide Dismutase
- Transfection
(methods)
- Ventricular Function, Left
- beta-Galactosidase
(genetics)
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