Comparison of HIV-1 and EIAV-based lentiviral vectors in corneal transduction.
Abstract: In this study we compare the ability of self-inactivating Human Immunodeficiency Virus 1 (HIV-1) and Equine Infectious Anaemia Virus (EIAV)-based vectors to mediate gene transfer to rabbit and human corneas and to a murine corneal endothelial cell line. Both vectors were pseudotyped with vesicular stomatitis virus-G (VSV-G) envelope and contained marker transgenes under the control of an internal CMV promoter. For specificity of action, the heterologous promoter in the EIAV-vector was exchanged for an inducible E-Selectin promoter, previously shown to regulate gene-expression in a plasmid system. We show that EIAV is more efficient than HIV in transducing human and rabbit corneal endothelial cells. Rabbit corneal endothelial cells are transduced in higher quantity than human corneal endothelial cells. In the inducible system, however, we detected impairment between the vector and its internal E-Selectin promoter. Instead of controlled transgene expression or silencing of promoter activity, the U3-modified long-terminal-repeats (LTR) impaired the conditional activity of the E-Selectin promoter. Significant transgene expression was seen without stimulation of the inducible promoter. We show efficient transduction by lentiviruses of a corneal endothelial cell line and of full thickness corneas from different species, confirming that those vectors would be appropriate tools for gene therapy of selected corneal diseases. However, the modification within the U3-LTR did not adequately allow regulated transgene expression. These findings have important implications for vector design for diagnostic or therapeutic opportunities.
Publication Date: 2005-01-21 PubMed ID: 15939034DOI: 10.1016/j.exer.2004.12.005Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research explores the efficiency of the Human Immunodeficiency Virus 1 (HIV-1) and the Equine Infectious Anaemia Virus (EIAV)-based vectors in gene transfer processes involving rabbit and human corneas and a particular mouse corneal endothelial cell line.
Experiment Setup
The researchers utilized both HIV-1 and EIAV vectors and modified them in order to examine their effectiveness in gene transfer processes. The vectors were pseudotyped with an envelope from the vesicular stomatitis virus-G (VSV-G) and contained particular marker transgenes governed by an internal CMV promoter.
- To ensure the desired specificity, they replaced the heterologous promoter in the EIAV vector with an inducible E-Selectin promoter, previously demonstrated to yield regulated gene-expression inside a plasmid system.
Findings and Observations
The team found and documented a range of results and observations, including:
- EIAV showing greater efficiency in transducing (introducing genes into) human and rabbit corneal endothelial cells over HIV-1.
- Rabbit corneal endothelial cells responded to transduction in larger quantities as compared to their human counterparts.
- A noticeable impairment between the EIAV-based vector and its internal E-Selectin promoter in the inducible system. Instead of the expected controlled transgene expression or promoter activity silencing, U3-modified long-terminal-repeats (LTR) restricted the conditional activity of the E-Selectin promoter. As a result, there was significant transgene expression, even without inducible promoter stimulation.
Conclusion and Implications
This study confirmed that lentiviruses can efficiently transduce a corneal endothelial cell line and full thickness corneas from different species. This suggests they can be used as an effective tool for gene therapy concerning certain corneal diseases. The study also highlighted areas that need operational improvements, specifically the U3-LTR’s modification that affected regulated transgene expression.
- These findings are crucial in understanding the potential applications and challenges of using lentiviral vectors in gene therapy and, more specifically, for addressing corneal diseases.
- This research carries significant implications for vector designs to be used in diagnostic or therapeutic opportunities.
Cite This Article
APA
Beutelspacher SC, Ardjomand N, Tan PH, Patton GS, Larkin DF, George AJ, McClure MO.
(2005).
Comparison of HIV-1 and EIAV-based lentiviral vectors in corneal transduction.
Exp Eye Res, 80(6), 787-794.
https://doi.org/10.1016/j.exer.2004.12.005 Publication
Researcher Affiliations
- Jefferiss Research Trust Laboratories, Wright-Fleming Institute, Faculty of Medicine, Division of Medicine, Department of GU Medicine, St Mary's Campus Medicine, Imperial College London, Norfolk Place, London W2 1PG, UK. s.beutelspacher@imperial.ac.uk
MeSH Terms
- Animals
- Cell Line
- Cornea / physiology
- E-Selectin / genetics
- Endothelium, Corneal / physiology
- Flow Cytometry / methods
- Gene Expression
- Genetic Vectors / genetics
- Green Fluorescent Proteins / genetics
- HIV-1 / genetics
- Humans
- Infectious Anemia Virus, Equine / genetics
- Liposomes
- Mice
- Mice, Inbred BALB C
- Promoter Regions, Genetic / genetics
- Rabbits
- Transduction, Genetic / methods
- Transfection / methods
- Transgenes / genetics
Citations
This article has been cited 12 times.- Munis AM. Gene Therapy Applications of Non-Human Lentiviral Vectors. Viruses 2020 Sep 29;12(10).
- Cavalieri V, Baiamonte E, Lo Iacono M. Non-Primate Lentiviral Vectors and Their Applications in Gene Therapy for Ocular Disorders. Viruses 2018 Jun 9;10(6).
- Mohan RR, Rodier JT, Sharma A. Corneal gene therapy: basic science and translational perspective. Ocul Surf 2013 Jul;11(3):150-64.
- Khan A, Fu H, Tan LA, Harper JE, Beutelspacher SC, Larkin DF, Lombardi G, McClure MO, George AJ. Dendritic cell modification as a route to inhibiting corneal graft rejection by the indirect pathway of allorecognition. Eur J Immunol 2013 Mar;43(3):734-46.
- Mohan RR, Tovey JC, Sharma A, Tandon A. Gene therapy in the cornea: 2005--present. Prog Retin Eye Res 2012 Jan;31(1):43-64.
- Fuchsluger TA, Jurkunas U, Kazlauskas A, Dana R. Anti-apoptotic gene therapy prolongs survival of corneal endothelial cells during storage. Gene Ther 2011 Aug;18(8):778-87.
- Fuchsluger TA, Jurkunas U, Kazlauskas A, Dana R. Corneal endothelial cells are protected from apoptosis by gene therapy. Hum Gene Ther 2011 May;22(5):549-58.
- Shinoda Y, Hieda K, Koyanagi Y, Suzuki Y. Efficient transduction of cytotoxic and anti-HIV-1 genes by a gene-regulatable lentiviral vector. Virus Genes 2009 Oct;39(2):165-75.
- Engler C, Kelliher C, Wahlin KJ, Speck CL, Jun AS. Comparison of non-viral methods to genetically modify and enrich populations of primary human corneal endothelial cells. Mol Vis 2009;15:629-37.
- Bustamante M, Tasinato A, Maurer F, Elkochairi I, Lepore MG, Arsenijevic Y, Pedrazzini T, Munier FL, Schorderet DF. Overexpression of a mutant form of TGFBI/BIGH3 induces retinal degeneration in transgenic mice. Mol Vis 2008 Jun 13;14:1129-37.
- Koh SW, Chandrasekara K, Abbondandolo CJ, Coll TJ, Rutzen AR. VIP and VIP gene silencing modulation of differentiation marker N-cadherin and cell shape of corneal endothelium in human corneas ex vivo. Invest Ophthalmol Vis Sci 2008 Aug;49(8):3491-8.
- Beutelspacher SC, Serbecic N, Tan P, McClure MO. [Comparison of several viral vectors for gene therapy of corneal endothelial cells]. Ophthalmologe 2005 Dec;102(12):1168-74.
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