In Vivo Gene Delivery by Lentiviral Vectors

Introduction

Successful gene therapy requires the efficient delivery and sustained expression of a therapeutic gene into the tissues of a human body. Most of the candidate tissues for therapeutic gene transfer are made of quiescent cells, such as from the brain, liver, and muscle. Thus, the optimal vector should infect nondividing cells, become stably associated with the genome of target cells, and support a high, steady-state level of transcription.^1,2 Like all vectors derived from retroviruses, lentiviral vectors integrate into the chromatin of target cells and do not transfer any viral genes. Both of these features are important for achieving sustained expression of the transgene. Moreover, lentiviral vectors infect nondividing cells, a feature sharply distinguishing them from simple or onco-retroviral vectors.³ Upon infection, retroviruses deliver a nucleoprotein complex that reverse transcribes the viral RNA and integrates the newly made DNA into the chromatin. The nucleoprotein complexes of onco-retroviruses are excluded by the nucleus, and they reach the chromatin only when the nuclear membrane is fragmented during mitosis. This explains the dependence of a productive onco-retroviral infection on cell division occurring shortly after viral entry.^4,5 In contrast, the nucleoprotein complexes of lentiviruses contain nuclear localization signals that mediate their active transport through the nucleopores during interphase. This explains the capacity of lentiviruses to infect macrophages, a nondividing cell type.^6-9

• References

• 1 Verma IM, Somia N. Gene therapy promises, problems and prospects. Nature 1997; 389: 239-242.
• 2 Anderson WF. Human gene therapy. Nature 1998; 392: 25-30.
• 3 Naldini L. Lentiviruses as gene transfer agents for delivery to non-dividing cells. Curr Opin Biotechnol 1998; 9: 457-463.
• 4 Miller DG, Adam MA, Miller AD. Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection. Mol Cell Biol 1990; 10: 4239-4242.
• 5 Roe T, Reynolds TC, Yu G, Brown PO. Integration of murine leukemia virus DNA depends on mitosis. EMBO J 1993; 12: 2099-2108.
• 6 Lewis PF, Emerman M. Passage through mitosis is required for oncoretroviruses but not for the human immunodeficiency virus. J Virol 1994; 68: 510.
• 7 Weinberg JB, Matthews TJ, Cullen BR, Malim MH. Productive human immunodeficiency virus type 1 (HIV-1) infection of non-proliferating human monocytes. J Exp Med 1991; 174: 1477.
• 8 Bukrinsky MI, Haggerty S, Dempsey MP, Sharova N, Adzhubel A, Spitz L, Lewis P, Goldfarb D, Emerman M, Stevenson M. A nuclear localization signal within HIV-1 matrix protein that governs infection of non-dividing cells. Nature 1993; 365: 666-669.
• 9 Gallay P, Swingler S, Aiken C, Trono D. HIV-1 infection of nondividing cells: C-terminal tyrosine phosphorylation of the viral matrix protein is a key regulator. Cell 1995; 80: 379-388.
• 10 Shimada T, Fujii H, Mitsuya A, Nienhuis W. Targeted and highly efficient gene transfer into CD4+ cells by a recombinant human immunodeficiency virus retroviral vector. J Clin Invest 1991; 88: 1043-1047.
• 11 Poznansky M, Lever A, Bergeron L, Haseltine W, Sodroski J. Gene transfer into human lymphocytes by a defective human immunodeficiency virus type 1 vector. J Virol 1991; 65: 532-536.
• 12 Buchschacher GLJ, Panganiban AT. Human immunodeficiency virus vectors for inducible expression of foreign genes. J Virology 1992; 66: 2731-2739.
• 13 Parolin C, Dorfman T, Palu G, Gottlinger H, Sodroski J. Analysis in human immunodeficiency virus type 1 vectors of cis-acting sequences that affect gene transfer into human lymphocytes. J Virol 1994; 68: 3888-3895.
• 14 Naldini L, Blömer U, Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 1996; 272: 263-267.
• 15 Burns JC, Friedmann T, Driever W, Burrascano M, Yee JK. Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and non-mammalian cells. Proc Natl Acad Sci USA 1993; 90: 8033-8037.
• 16 Akkina RK, Walton RM, Chen ML, Li Q-X, Planelles V, Chen ISY. High-efficiency gene transfer into CD34+ cells with a human immunodeficiency virus type 1-based retroviral vector pseudotyped with vesicular stomatitis virus envelope glycoprotein G. J Virol 1996; 70: 2581-2585.
• 17 Naldini L, Blömer U, Gage FH, Trono D, Verma IM. Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc Natl Acad Sci USA 1996; 93: 11382-11388.
• 18 Luciw PA. Human immunodeficiency viruses and their replication. In: Fields BN, Knipe DM, Howley PM, Chanock RM, Melnick JL, Monath TP, Roisman B, Straus SE. eds. Fields Virology. 3rd ed.. Philadelphia: Lippincott-Raven Publisher; 1996: 1881-1975.
• 19 Trono D. HIV accessory proteins: leading roles for the supporting cast. Cell 1995; 82: 189-192.
• 20 Zufferey R, Nagy D, Mandel RJ, Naldini L, Trono D. Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nature Biotech 1997; 15: 871-875.
• 21 Reiser J, Harmison G, Kluepfel-Stahl S, Brady RO, Karlsson S, Schubert M. Transduction of nondividing cells pseudotyped defective high-titer HIV type 1 particles. Proc Natl Acad Sci USA 1996; 93: 15266-15271.
• 22 Kim VN, Mitrophanous K, Kingsman SM, Kingsman AJ. Minimal requirement for a lentivirus vector based on human immunodeficiency virus type 1. J Virol 1998; 72: 811-816.
• 23 Wei P, Garber ME, Fang SM, Fischer WH, Jones KA. A novel CDK9-associated C-type cyclin interacts directly with HIV-1 Tat and mediates its high-affinity, loop-specific binding to TAR RNA. Cell 1998; 92: 451-462.
• 24 Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, Naldini L. A third generation lentiviral vector with a conditional packaging system. J Virol 1998; 72: 8463-8471.
• 25 Goh WC, Rogel ME, Kinsey CM, Michael SF, Fultz PN, Nowak MA, Hahn BH, Emerman M. HIV-1 Vpr increases viral expression by manipulation of the cell cycle: a mechanism for selection of Vpr in vivo. Nat Medicine 1998; 4: 65-71.
• 26 Zufferey R T, Dull RJ, Mandel A, Bukovsky D, Quiroz L, Naldini L, Trono D. Self-inactivating lentiviral vector for safe and efficient in vivo gene delivery. J Virol 1998; 72: 9873-9880
• 27 Miyoshi H, Blomer U, Takahashi M, Gage FH, Verma IM. Development of a self-inactivating lentivirus vector. J Virol 1998; 72: 8150-8157.
• 28 Bukovsky A, Song J-P, Naldini L. Interaction of HIV-derived vectors with wild-type virus in transduced cells. JOURNAL. In press.
• 29 Blömer U, Naldini L, Kafri T, Trono D, Verma IM, Gage FH. Highly efficient and sustained gene transfer in adult neurons with a lentivirus vector. J Virol 1997; 71: 6641-6649.
• 30 Miyoshi H., Takahashi M., Gage F.H., Verma I.M.. Stable and efficient gene transfer into the retina using an HIV-based lentiviral vertor. Proc Natl Acad Sci USA 1997; 94: 10319-10323.
• 31 Kafri T, Blömer U, Peterson DA, Gage FH, Verma I.M. Sustained expression of genes delivered directly into liver and muscle by lentiviral vectors. Nature Genet 1997; 17: 314-317.