Lentivirus Vectors

10 Key excerpts on "Lentivirus Vectors"

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  Guide To Human Gene Therapy, A

  • Roland W Herzog, Sergei Zolotukhin(Authors)
  • 2010(Publication Date)
  • World Scientific

    (Publisher)

  Chapter 4 Lentiviral Vectors Janka Mátrai, Marinee K. L. Chuah and Thierry VandenDriessche ∗ Lentiviral vectors represent some of the most promising vectors for gene therapy. They have emerged as potent and versatile tools for ex vivo or in vivo gene transfer into dividing and non-dividing cells. Lentiviral vec-tors can be pseudotyped with distinct viral envelopes that influence vector tropism and transduction efficiency. In addition, it is possible to redi-rect vector transduction and generate cell-type specific targetable vectors by incorporating cell-type specific ligands or antibodies into the vector envelope. Lentiviral vectors have been used to deliver genes into various tissues, including brain, retina and liver, by direct in vivo gene deliv-ery. Since they integrate stably into the target cell genome, they are ide-ally suited to deliver genes ex vivo into bona fide stem cells, particularly hematopoietic stem cells (HSCs), allowing for stable transgene expression upon hematopoietic reconstitution. Though there are some safety concerns regarding the risk of insertional mutagenesis, it is possible to minimize this risk by modifying the vector design or by employing integration-deficient lentiviral vectors which, in conjunction with zinc-finger nuclease tech-nology, allow for site-specific gene correction or addition in pre-defined chromosomal loci. Lentiviral gene transfer has provided efficient phe-notypic correction of diseases in mouse models paving the way towards clinical applications. 1. Basic Viral Biology Lentiviruses are enveloped RNA viruses that are surrounded by a lipid bilayer in which the envelope proteins are embedded. The main components ∗ Correspondence: Flanders Institute for Biotechnology,Vesalius Research Center, University of Leuven, Leuven, Belgium. E-mail: [email protected] 53 A Guide to Human Gene Therapy Fig. 4.1 A. Schematic representation of the HIV-1 virus.

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  Gene Therapy for Viral Infections

  • Patrick Arbuthnot(Author)
  • 2015(Publication Date)
  • Academic Press

    (Publisher)

  4.4. Lentiviral and Retroviral Vectors

  Retroviridae are the family of viruses to which lentiviruses and gammaretroviruses belong. Interestingly, retroviruses were among the earliest viral infectious agents to be discovered [86] . The equine infectious anemia virus, a lentivirus, and murine leukemia virus (MLV), a gammaretrovirus, were isolated as filterable infectious agents during the early 1900s. Subsequent outbreak of the HIV-1 pandemic, and the serious complications associated with the infection, provided enormous impetus to research members of the Retroviridae family (Chapter 9 ) and their development for gene transfer. A feature of lentiviruses and retroviruses is that they have RNA genomes that serve as templates for reverse transcription. After infection of cells, the reverse-transcribed DNA is integrated into the genome of host cells to form a provirus. This fundamental feature is crucial for achieving sustained transgene expression after transduction with the vectors.

  Vectors derived from lentiviruses have largely superseded their gammaretroviral counterparts because they have several advantages:

  1. Lentiviruses are capable of infecting nondividing or slowly dividing cells [87] , whereas retroviral vectors are only capable of transducing rapidly proliferating cells. This is as a result of the ability of the lentiviral preintegration complex (PIC) to traverse the intact nuclear membrane [88 ,89] .

  2. The considerable insights into the molecular biology of lentiviruses that have resulted from research on HIV-1 have led to improved understanding of the ways in which lentiviruses may be manipulated to improve the vectors’ properties. This has facilitated propagation of safer vectors with disabled viral transcriptional elements (reviewed in ref. [90] ). Improvement in the regulation of expression of transgenes and limiting interference with transcription of cellular genes after integration of the provirus has also resulted.

  3. The gene encoding the lentiviral reverse transcriptase may be conveniently mutated to render the vectors integration defective [91]

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  Molecular Virology

  • Moses P. Adoga(Author)
  • 2012(Publication Date)
  • IntechOpen

    (Publisher)

  However, the main disadvantage of this type of vector is its cell toxicity and low transduction efficiency. Currently, other variants that try to surpass these limitations have been developed. Lentivirus Vectors have a modest packaging capacity, induce minimal immunological response and can produce long-term transgene expression. In addition, envelope-engineered vectors can show broad cell tropism. On the other hand, these vectors show poor in vivo delivery and present the risk of insertional mutagenesis. However, as lentiviruses mostly transduce terminally differentiated cells, the risk of insertional mutagenesis is less important than observed for other retroviruses (Kaplitt & Pfaff, 1996; Davidson & Breakefield, 2003; Howarth et al., 2010; Snyder et al., 2010). 2. Therapeutic and research applications Viral vectors were originally developed as an alternative to transfection (see glossary ) of naked DNA for molecular genetics experiments. Compared to traditional methods such as calcium phosphate precipitation, transduction (see glossary ) can ensure that nearly 100% of cells are infected without severely affecting cell viability. Furthermore, some viruses integrate into the cell genome facilitating stable expression. Protein coding genes can be expressed using viral vectors, commonly to: 1. Increase concentration of a certain protein and study its function (over-expression studies); 2. Antagonize function of a certain protein (expression of dominant negative proteins and RNAi constructs); 3. Make the cell produce fluorescent indicator proteins (for example, EGFP or Ca2+ sensitive proteins). These may be used to monitor various variables within the living cells (tracing and in vivo imaging); 4. Control neuronal excitability using light-sensitive ion channels (optogenetics); 5.

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  Advanced Textbook On Gene Transfer, Gene Therapy And Genetic Pharmacology: Principles, Delivery And Pharmacological And Biomedical Applications Of Nucleotide-based Therapies

  Principles, Delivery and Pharmacological and Biomedical Applications of Nucleotide-Based Therapies

  • Daniel Scherman(Author)
  • 2013(Publication Date)
  • ICP

    (Publisher)

  PART II VECTORS AND GENE DELIVERY TECHNIQUES This page intentionally left blank This page intentionally left blank 117 8 γ -RETROVIRUS- AND LENTIVIRUS-DERIVED VECTORS FOR GENE TRANSFER AND THERAPY Caroline Duros a and Odile Cohen-Haguenauer a,b 8.1 Introduction Viral vectors can be manipulated in vitro to modify their genomes in order to insert a gene of interest. Intrinsic viral cycle properties, such as DNA importation into the nucleus and its expression, are therefore used to deliver the transgene to target cells. The major advantage of viral vectors derived from retroviruses is the integration of the transferred transgene into the chromosome of the host cell, which may enable long-lasting transgene expression, not only in the transduced target cell, but also in all offspring cells after cell division. 8.2 The Concept: Designing Retrovirus-Based Vectors 8.2.1 Starting From the Knowledge of Helper Retrovirus Biology Retroviruses are composed of oncoviruses, lentiviruses and spumaviruses. Oncoretroviruses can only infect dividing cells, because they require the disrup-tion of the nuclear membrane for the viral genome to get into the nucleus, whereas lentiviruses can also infect non-dividing cells. Oncoretroviruses and lentiviruses a Laboratory of Biotechnology and Applied Pharmacogenetics, CNRS UMR8 H3, Ecole Normale Supérieure de Cachan, France b Oncogenetics, Department of Clinical Oncology, Hôpital Saint-Louis, Unversité Paris7-Paris Diderot, Sorbonne Paris-Cité Paris, France Email: [email protected] 118 Advanced Textbook on Gene Transfer, Gene Therapy and Genetic Pharmacology are also first-choice vectors because they can be manipulated to obtain defective, non-replication-competent viruses in the target cells. Retroviruses are enveloped viruses with positive RNA, two identical RNAs being encapsidated in the viral particle.

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  Gene Transfer and Expression in Mammalian Cells

  • S.C. Makrides(Author)
  • 2003(Publication Date)
  • Elsevier Science

    (Publisher)

  Virus-based vectors for gene expression in mammalian cells: Lentiviruses

  Mehdi Gasmi1 , Flossie Wong-Staal2, * 1

  1 Ceregene, Inc., 9381 Judicial Drive #130, San Diego, CA 92121, USA. Tel.: +1 (858) 458-8828; Fax: +1 (858) 458-8801

  2 Department of Medicine MS0665, University of California San Diego, La Jolla, CA 92093, USA

  E-mail address: [email protected]

  Abstract Publisher Summary

  Lentiviruses, represented by the human immunodeficiency virus type 1 (HIV-1) are retroviruses that possess complex genomes and a finely regulated mode of replication. These viruses share the common feature of being able to infect post-mitotic cells of the monocyte/macrophage lineage; a phenomenon closely related to their biology and induced pathologies. Gene therapy vectors based on lentiviruses enable the generation of a gene delivery system that combines features of vectors derived from murine oncoretroviruses (i.e., large coding capacity and stable integration of the transgene into host-cell genetic material) and the capability of transducing non-dividing cells. As such, vectors derived from HIV-1 have been designed and the proof of principle of stable transgene delivery in non-dividing cells has been established in a wide variety of systems. The major hurdle that prevents the progression of the HIV-1-based vector system to the clinic for evaluation of therapeutic potential is the association of the parental virus with an incurable and still largely fatal disease in humans. An alternative approach to the construction of HIV-1-derived vectors, is the use of viruses derived from less pathogenic human immunodeficiency virus type 2 (HIV-2) and from simian immunodeficiency virus (SIV). Non-primate lentiviral vector systems derived from feline immunodeficiency virus (FIV), equine infectious anemia virus (EIAV), and visna/maedi virus (VMV) have also been described in the chapter. However, the impressive achievements accomplished with HIV-1-based vectors have largely overshadowed the characterization of these other lentiviral vector systems. Therefore, this chapter focuses on HIV-1 derived vectors to exemplify the basic design and improvements relevant to the development of a safe and efficient lentivirus-based gene delivery system.

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  Viral Vectors

  Gene Therapy and Neuroscience Applications

  • Michael G. Kaplitt, Arthur D. Loewy(Authors)
  • 1995(Publication Date)
  • Academic Press

    (Publisher)

  Genetics and Biology of Retroviral Vectors Rajat Bannerii Department of Molecular Biology MemoriaI-Sloan Kettering Cancer Center New York, New York In this chapter, retroviral vectors, based on the Moloney murine leukemia virus, are discussed as a method for delivering genes into target cells. This vector system offers the advantages of highly efficient gene transfer and stable gene expression in the target cell by virtue of the integration of the vector genome into the host chromosomal DNA. Other vector systems including adenovirus vectors, vaccinia virus vectors, and nonviral methods, such as lipo- somes, may allow only transient gene expression (Mulligan, 1993). Adeno- associated virus vectors do have the ability to stabily integrate into the host genome and will be discussed elsewhere (see Samulski, this volume). Helper- free retroviral vectors infect only once and do not spread in vivo, in contrast to other viral vector methods which employ attenuated viruses that may retain the ability to infect other cells. Retroviral vectors have a wide host range, both in terms of species and cell types (Hartley and Rowe, 1976; Rasheed et al., 1976; Luciw and Leung, 1992), but are limited to infecting only dividing cells (Varmus et al., 1977; Fritsch and Temin, 1977; Humphries et al., 1981). Overall, retroviral vectors offer an excellent gene delivery system available for stable gene transfer and expression. Retroviruses are single-stranded RNA viruses encoding a characteristic RNA-dependent DNA polymerase (reverse transcriptase, see Baltimore, 1970; Temin and Mizutani, 1970). They initially were identified as RNA tumor viruses by Peyton Rous, who was working at the Rockefeller Institute on a spontaneous chicken sarcoma (Rous, 1911). Since the discovery of the Rous sarcoma virus (RSV), many tumor causing retroviruses have been found in a variety of vertebrates including reptiles, birds, and mammals.

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  Gene and Cell Therapy

  Therapeutic Mechanisms and Strategies, Fourth Edition

  • Nancy Smyth Templeton(Author)
  • 2015(Publication Date)
  • CRC Press

    (Publisher)

  However, their ability to stably introduce a therapeutic gene in multiple different target cells and their favorable integration pattern predict that they might become powerful gene therapy tools in the near future. In parallel, safer LV vectors are being designed that will extend their applications to other genetic diseases and give hope to many patients suffering from these incurable diseases. REFERENCES 1. Sakuma, T., M.A. Barry, and Y. Ikeda, Lentiviral vectors: Basic to translational. Biochem J , 2012. 443 (3): 603–618. 2. Naldini, L., Lentiviruses as gene transfer agents for deliv-ery to non-dividing cells. Curr Opin Biotechnol , 1998. 9 (5): 457–463. 3. Vigna, E. et al., Robust and efficient regulation of transgene expression in vivo by improved tetracycline-dependent lenti-viral vectors. Mol Ther , 2002. 5 (3): 252–261. 4. Coiras, M. et al., Understanding HIV-1 latency provides clues for the eradication of long-term reservoirs. Nat Rev Microbiol , 2009. 7 (11): 798–812. 5. Frankel, A.D. and J.A. Young, HIV-1: Fifteen proteins and an RNA. Annu Rev Biochem , 1998. 67 : 1–25. 6. Vicenzi, E. and G. Poli, Novel factors interfering with human immunodeficiency virus-type 1 replication in vivo and in vitro. Tissue Antigens , 2013. 81 (2): 61–71. 7. Harris, R.S., J.F. Hultquist, and D.T. Evans, The restriction factors of human immunodeficiency virus. J Biol Chem , 2012. 287 (49): 40875–40883. 8. Metharom, P. et al., Novel bovine lentiviral vectors based on Jembrana disease virus. J Gene Med , 2000. 2 (3): 176–185. 9. Farley, D.C. et al., Development of an equine-tropic replication-competent lentivirus assay for equine infec-tious anemia virus-based lentiviral vectors. Hum Gene Ther Methods , 2012. 23 (5): 309–323. 10. Mendenhall, A. et al., Packaging HIV- or FIV-based lentivec-tor expression constructs and transduction of VSV-G pseudo-typed viral particles.

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  Gene Therapy

  Tools and Potential Applications

  • Francisco Martin Molina(Author)
  • 2013(Publication Date)
  • IntechOpen

    (Publisher)

  Gene trans‐ fer in humans using a conditionally replicating lentiviral vector. Proceedings of the National Academy of Sciences of the United States of America. 2006;103(46):17372–7. www.pnas.org/cgi/content/abstract/103/46/17372 (accessed 1 August 2012) Lentiviral Gene Therapy Vectors: Challenges and Future Directions http://dx.doi.org/10.5772/52534 317 Chapter 13 Lentiviral Vectors in Immunotherapy Ines Dufait, Therese Liechtenstein, Alessio Lanna, Roberta Laranga, Antonella Padella, Christopher Bricogne, Frederick Arce, Grazyna Kochan, Karine Breckpot and David Escors Additional information is available at the end of the chapter http://dx.doi.org/10.5772/50717 1. Introduction Genetic immunotherapy can be defined as a therapeutic approach in which therapeutic genes are introduced into defined target cell types to modulate immune responses. A major challenge for this therapeutic strategy is the delivery of these genes into target cells in an efficient, stable manner. Possibly one of the best systems to achieve this is the use of lentivi‐ ral vectors (lentivectors) as gene carriers, as they are capable of transducing both dividing and resting cells [1]. Lentivectors are mainly derived from the human immunodeficiency virus (HIV-1) genome, a member of the Retroviridae family. The defining characteristic of retroviruses is their ca‐ pacity to stably integrate their RNA genome into the host cell chromosomes, in the form of a cDNA copy (Figure 1). Therefore, retrovirus and Lentivirus Vectors have been used exten‐ sively in research since they are ideal gene carriers into target cells. Moreover, both retrovi‐ rus and Lentivirus Vectors have been successfully applied in human gene therapy for the treatment of several genetic/metabolic inherited diseases (Cartier et al, 2009; Cavazzana-Cal‐ vo et al, 2010; Gaspar et al, 2004; Grez et al, 2010; Ott et al, 2006; Thrasher et al, 2006).

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  Advances in Molecular Retrovirology

  • Shailendra K. Saxena(Author)
  • 2016(Publication Date)
  • IntechOpen

    (Publisher)

  Section 4 Retroviruses as Vectors in Gene Therapy Chapter 5 Retroviral Vectors in Gene Therapy Miroslava Matuskova and Erika Durinikova Additional information is available at the end of the chapter http://dx.doi.org/10.5772/61844 Abstract Several decades ago, the first retroviral vectors were constructed. They have been proved as delivery vehicles in basic and translational research; many of them were used in clini‐ cal trials in the treatment of genetic and immunologic disorders or malignancies to deliv‐ er therapeutic genes into target tissue. Gammaretroviral and lentiviral vectors are popular viral delivery vehicles; their ability to integrate into genome of the host cell ena‐ bles permanent genetic modification of the target cell and long-term expression of the transgene. Besides classical cancer gene therapy, they are used in cell-mediated cancer gene therapy in combination with mesenchymal stromal cells (MSC) or neural progeni‐ tors. Based on the promising preclinical studies, clinical trials with genetically engineered cell vehicles were initiated. Keywords: Retroviral vector, lentiviral vector, gene therapy 1. Introduction Besides negative and pathogenic attributes, viruses can also be beneficial when used as delivery vehicles in gene therapy. The advocates of viral vectors even claim that just viruses are the right tools for delivery of foreign genetic information into the cell because they have been evolving for this purpose for millions of years. Gene therapy can be defined as the delivery of nucleic acid into the cell for the purpose of acquiring new features or restoration of phys‐ iologic status. The idea that disorders can be treated by genes arose in the 1960s, when the mechanism of cell transformation by SV40 virus and papovaviruses was described [1]. Gene therapy enables modification of cell by the replacement of non-functional or missing gene, suppression of another gene, or induction of cell death as in the case of oncologic diseases.

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  Viral Gene Therapy

  • Ke Xu(Author)
  • 2011(Publication Date)
  • IntechOpen

    (Publisher)

  It was Brenner et al (Brenner et al. 1993) who demonstrated the proof-of-principle of γ -retroviral gene transfer in hematopoietic stem cells (HSCs). The first clinical trial using γ -retroviral vectors (RVs) was carried out by Anderson and colleagues to correct severe combined immunodeficiency (SCID) in 1991 (Anderson et al. 1990). The majority of clinical gene therapy trials today use γ -RVs, despite their relative low stability, the low titers and their inability to transduce quiescent cells. More importantly, worrisome incidents of RV induced insertional mutagenesis were reported (Pincha et al. 2010). Lentiviruses, which as γ -retroviruses are members of the Retroviridae , were suggested to be an attractive alternative, since they are capable of transducing both dividing and non-dividing cells (Bukrinsky et al. 1993; Lewis & Emerman 1994). Moreover, their integration into the host genome is, in contrast to γ -retroviruses, associated with lower genotoxicity (Montini et al. 2006). Dendritic Cells and Lentiviral Vectors: Mapping the Way to Successful Immunotherapy 317 At the end of the 1990s, the use of recombinant LVs was boosted, especially for transduction of non-dividing cells (Akkina et al. 1996; Naldini et al. 1996a; Naldini et al. 1996b; Reiser et al. 1996). Another 10 years later the first clinical trial using LV modified cells for the treatment of HIV infection was completed (Lu et al. 2004). 3.1 Development of recombinant lentiviral vectors Lentiviruses are characterized by a diploid 7-12 kb single stranded RNA genome with positive polarity that is reverse transcribed to double stranded DNA upon host cell entry ( Coffin 1997). Diploidy permits genetic recombination, which accounts partially for their success as procreators of the acquired immunodeficiency syndrome, a disease that develops by slowly affecting the immune systems’ function (lenti meaning slow). Lentiviruses include primate and non-primate retroviruses, e.g .

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