Zhou, Tian
(2011)
Effectiveness and safety of hydrodynamic gene delivery in animals with fibrotic liver.
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
Hydrodynamic gene delivery (HGD) has emerged as an effective and safe method for transfecting liver hepatocytes in vivo, and has potential for gene therapy of liver fibrosis. The objective of this study was to evaluate the effectiveness and safety of HGD using CCl4 induced fibrotic liver in rats as a model. I demonstrated that there is a progressive reduction of efficiency of HGD in rats with increasing severity of liver fibrosis. Using a reporter plasmid containing luciferase gene, we showed over 2,000-fold decrease in luciferase activity in the liver with advanced fibrosis compared to that of control animals. Reduction in reporter gene expression in fibrotic liver was correlated to lower copy number of plasmid DNA and less amount of luciferase mRNA in the liver. Microscopy analysis revealed significant accumulation of collagen fibers in the boundary of liver lobules and thickened hepatic sinusoidal endothelium. The morphological changes in fibrotic liver are associated with restriction of flow-through across the liver of DNA solution hydrodynamically injected and are responsible for the reduced gene delivery efficiency of the hydrodynamic procedure. Results from electrocardiogram and serum biochemistry show no difference between the control and fibrotic animals undergone HGD. These results suggest that the HGD is a safe method for gene transfer in animals with liver fibrosis but the effectiveness of gene delivery decreases with increase of severity of fibrosis. Future work should focus on adjustment of injection parameters (DNA dose, injection volume, injection speed) for optimal gene delivery to fibrotic liver.
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Details
| Item Type: |
University of Pittsburgh ETD
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| Status: |
Unpublished |
| Creators/Authors: |
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| ETD Committee: |
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| Date: |
12 August 2011 |
| Date Type: |
Completion |
| Defense Date: |
7 April 2011 |
| Approval Date: |
12 August 2011 |
| Submission Date: |
29 May 2011 |
| Access Restriction: |
5 year -- Restrict access to University of Pittsburgh for a period of 5 years. |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Pharmacy > Pharmaceutical Sciences |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
hydrodynamic delivery; liver fibrosis; transgene expression |
| Other ID: |
http://etd.library.pitt.edu/ETD/available/etd-05292011-230802/, etd-05292011-230802 |
| Date Deposited: |
10 Nov 2011 19:46 |
| Last Modified: |
15 Nov 2016 13:44 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/7975 |
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