A method of gene transfer for the treatment of ischemic heart failure is described using a swine model of myocardial infarction. Our simple and reproducible method enables us to readily evaluate the efficacy of various gene transfers with a very simple and reproducible way.
A subscription to JoVE is required to view this content. Sign in or start your free trial.
Method Article
Gene Transfer for Ischemic Heart Failure in a Preclinical Model
In This Article
Summary
Abstract
Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety.
Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential.
In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates.
Protocol
1. Myocardial infarct creation
- Female Yorkshire pigs (20kg) from Animal Biotech Industries (Doylestown, PA) are pre-medicated using Telazol (tiletamine/zolazepam) (8.0 mg/kg) and Buprenorphine (0.6 mg).
- Animals are intubated and then ventilated with 100% oxygen. General anesthesia is maintained with Propofol (5-8mg/kg/hr) throughout the procedure.
- The femoral site is prepared with 70% isopropyl alcohol followed by povidone iodine. A percutaneous puncture provides access to the artery and for sheath placement. After sheath insertion, 3000-5000 IU of heparin is administered IV to maintain an activated coagulation time of 250-300 seconds.<....
Access restricted. Please log in or start a trial to view this content.
Discussion
Among the various causes of heart failure, ischemia is by far the most frequent in etiology. To overcome this serious disease, various therapeutic approaches are being developed. Careful evaluations of therapies are required before these promising therapies are can be applied to the clinics. Therefore, establishment of an ischemic heart failure model that represents the clinical conditions of patients is critical to development of heart failure therapies. Our swine myocardial infarct model meets these translational requ.......
Access restricted. Please log in or start a trial to view this content.
Disclosures
All authors have no commercial associations that might pose as a conflict of interest in connection with the manuscript.
Acknowledgements
This work is supported by Leducq Foundation through the Caerus network (RJH), by NIH R01 HL093183, HL088434, HL071763, HL080498, HL083156, and P20HL100396 (RJH). DL was supported by the German Research Foundation. We would like to thank Lauren Leonardson for her technical assistance in this project.
....Access restricted. Please log in or start a trial to view this content.
Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Telazol | Fort Dodge Animal Health | 0856-9050-49 | |
| Propofol | APP Pharmaceuticals | 63323-270-20 | |
| Heparin | APP Pharmaceuticals | 63323-540-11 | |
| Amiodarone | APP Pharmaceuticals | 63323-616-03 | |
| Atropin Sulfate | Med-Pharmex | 54925-063-10 | |
| Potassium acetate | American Regent Inc. | 2053-25 | |
| Nitroglycerine | American Regent Inc. | 4810-25 | |
| Salix (Furosemide) | Intervet Inc. | 710461 | |
| 5 Fr Hockey stick catheter | Cordis | 556-278-0L | |
| 7 Fr Hockey stick catheter | Cordis | 778-278-00 | |
| Coronary wire | Abbott Laboratories | 1044588 | |
| Coronary balloon | Abbott Laboratories | 1009447-08 | |
| 8Fr sheath | Terumo Medical Corp. | RSS801 |
References
- Perez de Prado, A. Closed-chest experimental porcine model of acute myocardial infarction-reperfusion. J Pharmacol Toxicol Methods. 60, 301-306 (2009).
- Angeli, F. S. Left ventricular remodeling after myocardi....
Access restricted. Please log in or start a trial to view this content.
Reprints and Permissions
