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Experimental eye research2005; 80(6); 787-794; doi: 10.1016/j.exer.2004.12.005

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.
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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

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

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

Experimental eye research
ISSN: 0014-4835
NlmUniqueID: 0370707
Country: England
Language: English
Volume: 80
Issue: 6
Pages: 787-794

Researcher Affiliations

Beutelspacher, Sven Christoph
  • 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
Ardjomand, Navid
    Tan, Peng Hong
      Patton, Gillian Sarah
        Larkin, D Frank P
          George, Andrew J T
            McClure, Myra O

              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.
              1. Munis AM. Gene Therapy Applications of Non-Human Lentiviral Vectors. Viruses 2020 Sep 29;12(10).
                doi: 10.3390/v12101106pubmed: 33003635google scholar: lookup
              2. 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).
                doi: 10.3390/v10060316pubmed: 29890733google scholar: lookup
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                doi: 10.1002/eji.201242914pubmed: 23212959google scholar: lookup
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                doi: 10.1016/j.preteyeres.2011.09.001pubmed: 21967960google scholar: lookup
              6. 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.
                doi: 10.1038/gt.2011.20pubmed: 21412281google scholar: lookup
              7. 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.
                doi: 10.1089/hum.2010.079pubmed: 21158568google scholar: lookup
              8. 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.
                doi: 10.1007/s11262-009-0382-xpubmed: 19554442google scholar: lookup
              9. 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.
                pubmed: 19347048
              10. 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.
                pubmed: 18568131
              11. 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.
                doi: 10.1167/iovs.07-1543pubmed: 18441300google scholar: lookup
              12. 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.
                doi: 10.1007/s00347-005-1230-6pubmed: 15886987google scholar: lookup
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